Saeid Ghavami

The inactivation of programmed cell death has profound effects not only on the development but also on the overall integrity of multicellular organisms. Beside developmental abnormalities, it may lead to tumorigenesis, autoimmunity, and other serious health problems. Deregulated apoptosis may also be the leading cause of cancer therapy chemoresistance. Caspase family of cysteinyl-proteases plays the key role in the initiation and execution of programmed cell death. This review gives an overview of the role of caspases, their natural modulators like IAPs, FLIPs, and Smac/Diablo in apoptosis and upon inactivation, and also in cancer development. Besides describing the basic mechanisms governing programmed cell death, a large part of this review is dedicated to previous studies that were focused on screening tumours for mutations within caspase genes as well as their regulators. The last part of this review discusses several emerging treatments that involve modulation of caspases and their regulators. Thus, we also highlight caspase cascade modulating experimental anticancer drugs like cFLIP-antagonist CDDO-Me; cIAP1 antagonists OSU-03012 and ME-BS; and XIAP small molecule antagonists 1396-11, 1396-12, 1396-28, triptolide, AEG35156, survivin/Hsp90 antagonist shephedrin, and some of the direct activators of procaspase-3.

In December 2019, a novel SARS-CoV-2 coronavirus emerged, causing an outbreak of life-threatening pneumonia in the Hubei province, China, and has now spread worldwide, causing a pandemic. The urgent need to control the disease, combined with the lack of specific and effective treatment modalities, call for the use of FDA-approved agents that have shown efficacy against similar pathogens. Chloroquine, remdesivir, lopinavir/ritonavir or ribavirin have all been successful in inhibiting SARS-CoV-2 in vitro. The initial results of a number of clinical trials involving various protocols of administration of chloroquine or hydroxychloroquine mostly point towards their beneficial effect. However, they may not be effective in cases with persistently high viremia, while results on ivermectin (another antiparasitic agent) are not yet available. Interestingly, azithromycin, a macrolide antibiotic in combination with hydroxychloroquine, might yield clinical benefit as an adjunctive. The results of clinical trials point to the potential clinical efficacy of antivirals, especially remdesivir (GS-5734), lopinavir/ritonavir, and favipiravir. Other therapeutic options that are being explored involve meplazumab, tocilizumab, and interferon type 1. We discuss a number of other drugs that are currently in clinical trials, whose results are not yet available, and in various instances we enrich such efficacy analysis by invoking historic data on the treatment of SARS, MERS, influenza, or in vitro studies. Meanwhile, scientists worldwide are seeking to discover novel drugs that take advantage of the molecular structure of the virus, its intracellular life cycle that probably elucidates unfolded-protein response, as well as its mechanism of surface binding and cell invasion, like angiotensin converting enzymes-, HR1, and metalloproteinase inhibitors.

Cellular homeostasis and adaptation to various environmental conditions are importantly regulated by the sophisticated mechanism of autophagy and its crosstalk with Wnt signaling and other developmental pathways. Both autophagy and Wnt signaling are involved in embryogenesis and differentiation. Autophagy is responsible for degradation and recycling of cytosolic materials by directing them to lysosomes through the phagophore compartment. A dual feedback mechanism regulates the interface between autophagy and Wnt signaling pathways. During nutrient deprivation, β-catenin and Dishevelled (essential Wnt signaling proteins) are targeted for autophagic degradation by LC3. When Wnt signaling is activated, β-catenin acts as a corepressor of one of the autophagy proteins, p62. In contrast, another key Wnt signaling protein, GSK3β, negatively regulates the Wnt pathway and has been shown to induce autophagy by phosphorylation of the TSC complex. This article reviews the interplay between autophagy and Wnt signaling, describing how β-catenin functions as a key cellular integration point coordinating proliferation with autophagy, and it discusses the clinical importance of the crosstalk between these mechanisms.

Protein homeostasis is a vital process for cell function and, therefore, disruption of the molecular mechanisms involved in this process, such as autophagy, may contribute to neurodegenerative diseases (NDs). Apart from autophagy disruption, excess oxidative stress and endoplasmic reticulum (ER) stress are additional main molecular mechanisms underlying neurodegeneration, leading to protein aggregation, and mitochondrial dysfunction. Notably, these primary molecular processes are interconnected pathways, which have synergistic effects on each other. Therefore, we propose that targeting of the crosstalk between autophagy, oxidative stress and ER stress simultaneously may play a critical role in healing NDs. NeuroNanoTechnology, as a revolutionized approach, in combination with an in-silico strategy, holds great promise for developing de-novo structures for targeting and modulating neuro-molecular pathways. Accordingly, this review outlines the contributions of autophagy, oxidative stress, and ER stress in neurodegenerative conditions along with a particular focus on the crosstalk among these pathways. Furthermore, we provide a comprehensive discussion on the potential of nanomaterials to target this crosstalk and suggest this potential as a promising opportunity in neuroprotection.

Metastasis consists of hallmark events, including Epithelial-Mesenchymal Transition (EMT), angiogenesis, initiation of inflammatory tumor microenvironment, and malfunctions in apoptosis. Autophagy is known to play a pivotal role in the metastatic process. Autophagy has pulled researchers towards it in recent times because of its dual role in the maintenance of cancer cells. Evidence states that cells undergoing EMT need autophagy in order to survive during migration and dissemination. Additionally, it orchestrates EMT markers in certain cancers. On the other side of the coin, autophagy plays an oncosuppressive role in impeding early metastasis. This review aims to project the interrelationship between autophagy and EMT. Targeting EMT via autophagy as a useful strategy is discussed in this review. Furthermore, for the first time, we have covered the possible reciprocating roles of EMT and autophagy and its consequences in cancer metastasis.

The statin drugs ('statins') potently inhibit hydroxymethylglutaryl-coenzyme A (HMG-CoA) reductase by competitively blocking the active site of the enzyme. Statins decrease de novo cholesterol biosynthesis and thereby reduce plasma cholesterol levels. Statins exhibit "pleiotropic" properties that are independent of their lipid-lowering effects. For example, preclinical evidence suggests that statins inhibit tumor growth and induce apoptosis in specific cancer cell types. Furthermore, statins show chemo-sensitizing effects by impairing Ras family GTPase signaling. However, whether statins have clinically meaningful anti-cancer effects remains an area of active investigation. Both preclinical and clinical studies on the potential mechanisms of action of statins in several cancers have been reviewed in the literature. Considering the contradictory data on their efficacy, we present an up-to-date summary of the pleiotropic effects of statins in cancer therapy and review their impact on different malignancies. We also discuss the synergistic anti-cancer effects of statins when combined with other more conventional anti-cancer drugs to highlight areas of potential therapeutic development.

Lipotoxicity is a form of cellular stress caused by the accumulation of lipids resulting in mitochondrial dysfunction and insulin resistance in muscle. Previously, we demonstrated that the mitophagy receptor BNIP3L/Nix is responsive to lipotoxicity and accumulates in response to a high-fat (HF) feeding. To provide a better understanding of this observation, we undertook gene expression array and shot-gun metabolomics studies in soleus muscle from rodents on an HF diet. Interestingly, we observed a modest reduction in several autophagy-related genes. Moreover, we observed alterations in the fatty acyl composition of cardiolipins and phosphatidic acids. Given the reported roles of these phospholipids and BNIP3L in mitochondrial dynamics, we investigated aberrant mitochondrial turnover as a mechanism of impaired myocyte insulin signaling. In a series of gain-of-function and loss-of-function experiments in rodent and human myotubes, we demonstrate that BNIP3L accumulation triggers mitochondrial depolarization, calcium-dependent activation of DNM1L/DRP1, and mitophagy. In addition, BNIP3L can inhibit insulin signaling through activation of MTOR-RPS6KB/p70S6 kinase inhibition of IRS1, which is contingent on phosphatidic acids and RHEB. Finally, we demonstrate that BNIP3L-induced mitophagy and impaired glucose uptake can be reversed by direct phosphorylation of BNIP3L by PRKA/PKA, leading to the translocation of BNIP3L from the mitochondria and sarcoplasmic reticulum to the cytosol. These findings provide insight into the role of BNIP3L, mitochondrial turnover, and impaired myocyte insulin signaling during an overfed state when overall autophagy-related gene expression is reduced. Furthermore, our data suggest a mechanism by which exercise or pharmacological activation of PRKA may overcome myocyte insulin resistance.Abbreviations: BCL2: B cell leukemia/lymphoma 2; BNIP3L/Nix: BCL2/adenovirus E1B interacting protein 3-like; DNM1L/DRP1: dynamin 1-like; FUNDC1: FUN14 domain containing 1; IRS1: insulin receptor substrate 1; MAP1LC3A/LC3: microtubule-associated protein 1 light chain 3 alpha; MFN1: mitofusin 1; MFN2: mitofusin 2; MTOR: mechanistic target of rapamycin kinase; OPA1: OPA1 mitochondrial dynamin like GTPase; PDE4i: phosphodiesterase 4 inhibitor; PLD1: phospholipase D1; PLD6: phospholipase D family member 6; PRKA/PKA: protein kinase, AMP-activated; PRKCD/PKCδ: protein kinase C, delta; PRKCQ/PKCθ: protein kinase C, theta; RHEB: Ras homolog enriched in brain; RPS6KB/p70S6K: ribosomal protein S6 kinase; SQSTM1/p62: sequestosome 1; YWHAB/14-3-3β: tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein beta.

The coronavirus disease 2019 (COVID-19) pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection has profoundly affected the lives of millions of people. To date, there is no approved vaccine or specific drug to prevent or treat COVID-19, while the infection is globally spreading at an alarming rate. Because the development of effective vaccines or novel drugs could take several months (if not years), repurposing existing drugs is considered a more efficient strategy that could save lives now. Statins constitute a class of lipid-lowering drugs with proven safety profiles and various known beneficial pleiotropic effects. Our previous investigations showed that statins have antiviral effects and are involved in the process of wound healing in the lung. This triggered us to evaluate if statin use reduces mortality in COVID-19 patients.After initial recruitment of 459 patients with COVID-19 (Shiraz province, Iran) and careful consideration of the exclusion criteria, a total of 150 patients, of which 75 received statins, were included in our retrospective study. Cox proportional-hazards regression models were used to estimate the association between statin use and rate of death. After propensity score matching, we found that statin use appeared to be associated with a lower risk of morbidity [HR = 0.85, 95% CI = (0.02, 3.93), P = 0.762] and lower risk of death [(HR = 0.76; 95% CI = (0.16, 3.72), P = 0.735)]; however, these associations did not reach statistical significance. Furthermore, statin use reduced the chance of being subjected to mechanical ventilation [OR = 0.96, 95% CI = (0.61-2.99), P = 0.942] and patients on statins showed a more normal computed tomography (CT) scan result [OR = 0.41, 95% CI = (0.07-2.33), P = 0.312].Although we could not demonstrate a significant association between statin use and a reduction in mortality in patients with COVID19, we do feel that our results are promising and of clinical relevance and warrant the need for prospective randomized controlled trials and extensive retrospective studies to further evaluate and validate the potential beneficial effects of statin treatment on clinical symptoms and mortality rates associated with COVID-19.

Alzheimer's disease (AD) is one of the biggest human health threats due to increases in aging of the global population. Unfortunately, drugs for treating AD have been largely ineffective. Interestingly, downregulation of macroautophagy (autophagy) plays an essential role in AD pathogenesis. Therefore, targeting autophagy has drawn considerable attention as a therapeutic approach for the treatment of AD. However, developing new therapeutics is time-consuming and requires huge investments. One of the strategies currently under consideration for many diseases is “drug repositioning” or “drug repurposing”. In this comprehensive review, we have provided an overview of the impact of autophagy on AD pathophysiology, reviewed the therapeutics that upregulate autophagy and are currently used in the treatment of other diseases, including cancers, and evaluated their repurposing as a possible treatment option for AD. In addition, we discussed the potential of applying nano-drug delivery to neurodegenerative diseases, such as AD, to overcome the challenge of crossing the blood brain barrier and specifically target molecules/pathways of interest with minimal side effects.

Spermatogenesis is a complex process in the testis and is a cornerstone of male infertility. The abundance of unsaturated fatty acid and high cell division rate make male germs cells prone to DNA deterioration. ROS-mediated oxidative stress triggers DNA damage, autophagy, and apoptosis in male germ cells, which are critical causative factors that lead to male infertility. The complex connection and molecular crosstalk between apoptosis and autophagy is seen at multifaceted levels that interconnect the signaling pathways of these two processes. Multilevel interaction between apoptosis and autophagy is a seamless state of survival and death in response to various stressors. Interaction between multiple genes and proteins such as the mTor signaling pathway, Atg12 proteins, and the death adapter proteins, such as Beclin 1, p53, and Bcl-2 family proteins, validates such a link between these two phenomena. Testicular cells being epigenetically different from somatic cells, undergo numerous significant epigenetic transitions, and ROS modulates the epigenetic framework of mature sperm. Epigenetic deregulation of apoptosis and autophagy under oxidative stress conditions can cause sperm cell damage. The current review recapitulates the current role of prevailing stressors that generate oxidative stress leading to the induction of apoptosis and autophagy in the male reproductive system. Considering the pathophysiological consequences of ROS-mediated apoptosis and autophagy, a combinatorial approach, including apoptosis inhibition and autophagy activation, should be implemented as a therapeutic strategy to treat male idiopathic infertility. Understanding the crosslink between apoptosis and autophagy under stress conditions in male germ cells may play an essential role in developing therapeutic strategies to treat infertility.

Liver fibrosis is characterized by the excessive deposition and accumulation of extracellular matrix components, mainly collagens, and occurs in response to a broad spectrum of triggers with different etiologies. Under stress conditions, autophagy serves as a highly conserved homeostatic system for cell survival and is importantly involved in various biological processes. Transforming growth factor-β1 (TGF-β1) has emerged as a central cytokine in hepatic stellate cell (HSC) activation and is the main mediator of liver fibrosis. A growing body of evidence from preclinical and clinical studies suggests that TGF-β1 regulates autophagy, a process that affects various essential (patho)physiological aspects related to liver fibrosis. This review comprehensively highlights recent advances in our understanding of cellular and molecular mechanisms of autophagy, its regulation by TGF-β, and the implication of autophagy in the pathogenesis of progressive liver disorders. Moreover, we evaluated crosstalk between autophagy and TGF-β1 signalling and discussed whether simultaneous inhibition of these pathways could represent a novel approach to improve the efficacy of anti-fibrotic therapy in the treatment of liver fibrosis.

The study of autophagy ('self-eating'), a fundamental cell fate pathway involved in physiological and pathological subcellular processes, opens a new frontier in the continuous search for novel therapies for human asthma. Asthma is a complex syndrome with different disease phenotypes. Autophagy plays a central role in cell physiology, energy and metabolism, and cell survival. Autophagy's hallmark is the formation of double-membrane autophagic autophagosomes, and this process is operational in airway epithelial and mesenchymal cells in asthma. Genetic associations between autophagy genes and asthma have been observed including single nucleotide polymorphisms in Atg5 which correlate with reduced lung function. Immune mechanisms important in asthma such as Th2 cells and eosinophils also manifest autophagy. Lastly, we address the role of autophagy in extracellular matrix deposition and fibrosis in asthmatic airways remodeling, a pathologic process still without effective therapy, and discuss potential pharmacologic inhibitors. We end by offering two opposing but plausible hypotheses as to how autophagy may be directly involved in airway fibrosis.

Magnetic nanomaterials (MNMs) have gained great interest from different fields of study ranging from wastewater treatment to (bio)sensor development, taking advantage of both nanoscale size (allowing high surface-to-volume ratios) and the opportunity for magnetic manipulation. These materials can be surface-modified with antibodies, oligonucleotides, and aptamers to enable selective binding with target viruses or their biomarkers in biological samples. Using an external magnetic field, MNM-virus/biomarker complexes can be effectively isolated for further analysis. In some cases, the role of MNMs is not limited to simply serve as magnetic sorbents for extraction purposes as they have become an active part of some emerging detection processes (e.g., the use of magnetoresistive sensors). The combined application of MNMs with microfluidics for virus detection provides promising avenues for diagnostic tests that are of lower cost, require less time, and have higher specificity and sensitivity over conventional tests. This review focuses on the different approaches of virus detection using MNMs integrated in microfluidic chips. We will discuss recent research findings and provide insights and future perspectives for the development of low-cost and effective COVID-19 diagnostics tests.

The development of novel therapeutic approaches is necessary to manage gastrointestinal cancers (GICs). Considering the effective molecular mechanisms involved in tumor growth, the therapeutic response is pivotal in this process. Autophagy is a highly conserved catabolic process that acts as a double-edged sword in tumorigenesis and tumor inhibition in a context-dependent manner. Depending on the stage of malignancy and cellular origin of the tumor, autophagy might result in cancer cell survival or death during the GICs' progression. Moreover, autophagy can prevent the progression of GIC in the early stages but leads to chemoresistance in advanced stages. Therefore, targeting specific arms of autophagy could be a promising strategy in the prevention of chemoresistance and treatment of GIC. It has been revealed that autophagy is a cytoplasmic event that is subject to transcriptional and epigenetic regulation inside the nucleus. The effect of epigenetic regulation (including DNA methylation, histone modification, and expression of non-coding RNAs (ncRNAs) in cellular fate is still not completely understood. Recent findings have indicated that epigenetic alterations can modify several genes and modulators, eventually leading to inhibition or promotion of autophagy in different cancer stages, and mediating chemoresistance or chemosensitivity. The current review focuses on the links between autophagy and epigenetics in GICs and discusses: 1) How autophagy and epigenetics are linked in GICs, by considering different epigenetic mechanisms; 2) how epigenetics may be involved in the alteration of cancer-related phenotypes, including cell proliferation, invasion, and migration; and 3) how epidrugs modulate autophagy in GICs to overcome chemoresistance.

Autophagy is a very well-coordinated intracellular process that maintains cellular homeostasis under basal conditions by removing unnecessary or dysfunctional components through orderly degradation and recycling. Under pathological conditions, defects in autophagy have been linked to various human disorders, including neurodegenerative disorders and cancer. The role of autophagy in stem cell proliferation, differentiation, self-renewal, and senescence is well documented. Additionally, cancer stem cells (CSCs) play an important role in tumorigenesis, metastasis and tumor relapse and several studies have suggested the involvement of autophagy in the maintenance and invasiveness of CSCs. Hence, considering the modulation of autophagy in normal and cancer stems cells as a therapeutic approach can lead to the development or improvement of regenerative and anti-cancer therapies. Accordingly, modulation of autophagy can be regarded as a target for stem cell-based therapy of diseases with abnormal levels of autophagy. This article is focused on understanding the role of autophagy in stem cell homeostasis with an emphasis on the therapeutic potential of targeting autophagy for future therapies.

Proteasome subunit alpha type-2 (PSMA2) is a critical component of the 20S proteasome, which is the core particle of the 26S proteasome complex and is involved in cellular protein quality control by recognizing and recycling defective proteins. PSMA2 expression dysregulation has been detected in different human diseases and viral infections. No study yet has reported PSMA2 knockdown (KD) effects on the cellular proteome. Methods: We used SOMAScan, an aptamer-based multiplexed technique, to measure >1300 human proteins to determine the impact of PSMA2 KD on A549 human lung epithelial cells. Results: PSMA2 KD resulted in significant dysregulation of 52 cellular proteins involved in different bio-functions, including cellular movement and development, cell death and survival, and cancer. The immune system and signal transduction were the most affected cellular functions. PSMA2 KD caused dysregulation of several signaling pathways involved in immune response, cytokine signaling, organismal growth and development, cellular stress and injury (including autophagy and unfolded protein response), and cancer responses. Conclusions: In summary, this study helps us better understand the importance of PSMA2 in different cellular functions, signaling pathways, and human diseases.

The COVID-19 pandemic remains an emerging public health crisis with serious adverse effects. The disease is caused by severe acute respiratory syndrome coronavirus-2 (SARS-CoV--2) infection, targeting angiotensin-converting enzyme-2 (ACE2) receptor for cell entry. However, changes in the renin-angiotensin system (RAS) balance alter an individual's susceptibility to COVID-19 infection. We aimed to evaluate the association between AGT rs699 C > T, ACE rs4646994 I/D, and AGTR1 rs5186 C > A variants and the risk of COVID-19 infection and the severity in a sample of the southeast Iranian population.A total of 504 subjects, including 258 COVID-19 positives, and 246 healthy controls, were recruited. Genotyping of the ACE gene rs4646994, and AGT rs699, and AGTR1 rs5186 polymorphisms was performed by polymerase chain reaction (PCR) and PCR-restriction fragment length polymorphism (PCR-RFLP), respectively.Our results showed that the II genotype of ACE rs4646994 and the I allele decreased the risk of COVID-19 infection. Moreover, we found that the TC genotype and C allele of AGT rs699 increased the risk of COVID-19 infection. The AGTR1 rs5186 was not associated with COVID-19 infection. Also, we did not find any association between these polymorphisms and the severity of the disease. However, we found a significantly higher age and prevalence of diabetes and hypertension in patients with severe disease than a non-severe disease.These findings suggest that ACE rs4646994 and AGT rs699 polymorphisms increase the risk of COVID-19 infection in a southeast Iranian population.

Lung cells are constantly exposed to various internal and external stressors that disrupt protein homeostasis. To cope with these stimuli, cells evoke a highly conserved adaptive mechanism called the unfolded protein response (UPR). UPR stressors can impose greater protein secretory demands on the endoplasmic reticulum (ER), resulting in the development, differentiation, and survival of these cell types to meet these increasing functional needs. Dysregulation of the UPR leads to the development of the disease. The UPR and ER stress are involved in several human conditions, such as chronic inflammation, neurodegeneration, metabolic syndrome, and cancer. Furthermore, potent and specific compounds that target the UPR pathway are under development as future therapies. The focus of this review is to thoroughly describe the effects of both internal and external stressors on the ER in asthma. Furthermore, we discuss how the UPR signaling pathway is activated in the lungs to overcome cellular damage. We also present an overview of the pathogenic mechanisms, with a brief focus on potential strategies for pharmacological interventions.

MicroRNAs (miRNAs or miRs) are a family of small non-coding RNAs that function as oncogenes or tumor suppressor genes. Recent evidence suggests that the pri-miR-34b/c rs4938723 variant is associated with the development of cancer. At present, there is an inconsistent association between the single-nucleotide polymorphism in pri-miR-34b/c and cancer in the limited studies. The present study is a case-control investigation, with 263 breast cancer (BC) patients and 221 control women, which examined the potential association of the pri-miR-34b/c rs4938723 polymorphisms with BC susceptibility. The polymorphisms were genotyped by the polymerase chain reaction restriction fragment length polymorphism method. No significant association between the pri-miR-34b/c rs4938723 variant and BC was identified [TC vs. TT: Odds ratio (OR), 0.87; 95% confidence interval (CI), 0.60-1.26; P=0.506; CC vs. TT: OR, 1.22; 95% CI, 0.61-2.47; P=0.600; TC+CC vs. TT: OR, 0.91; 95% CI, 0.64-1.31; P=0.648; CC vs. TT+TC: OR, 1.32; 95% CI, 0.67-2.59; P=0.498; C vs. T: OR, 0.99; 95% CI, 0.75-1.31; P=0.986]. However, a significant association was observed between the pri-miR-34b/c rs4938723 genotypes and clinicopathological characteristics, such a grade, progesterone receptor and human epidermal growth factor receptor 2 status were observed (P<0.05). These findings suggest that the pri-miR-34b/c rs4938723 variant may not be a risk factor for the development of BC.

The liver is continuously exposed to a large antigenic load that includes pathogens, toxins, tumor cells and dietary antigens. Amongst the hepatitis viruses, only hepatitis B virus (HBV) and hepatitis C virus (HCV) cause chronic hepatitis, which can progress to cirrhosis and hepatocellular carcinoma. Of the different antiviral defense systems employed by the tissue, apoptosis significantly contributes to the prevention of viral replication, dissemination, and persistence. Loss of tolerance to the liver autoantigens may result in autoimmune hepatitis (AIH). This review outlines the recent findings that highlight the role and mechanisms of apoptotic processes in the course of liver diseases. Among factors that contribute to liver pathology, we discuss the role of tumor necrosis factor (TNF)-alpha, HBx, ds-PKR, TRAIL, FasL, and IL-1alpha. Since TNF and FasL-induced hepatocyte apoptosis is implicated in a wide range of liver diseases, including viral hepatitis, alcoholic hepatitis, ischemia/reperfusion liver injury, and fulminant hepatic failure, these items will be discussed in greater detail in this review. We also highlight some recent discoveries that pave the way for the development of new therapeutic strategies by protecting hepatocytes (for example by employing Bcl-2, Bcl-XL or A1/Bfl-1, IAPs, or synthetic caspase inhibitors), or by the induction of apoptosis in stellate cells. The assessment of the severity of liver disease, as well as monitoring of patients with chronic liver disease, remains a major challenge in clinical hepatology practice. Therefore, a separate chapter is devoted to a novel cytochrome c-based method useful for the diagnosis and monitoring of fulminant hepatitis.

The recent SARS-CoV-2 pandemic poses one of the greatest challenges to modern medicine. Therefore, identification of new therapeutic strategies seems essential either based on novel vaccines or drugs or simply repurposing existing drugs. Notably, due to their known safety profile, repurposing of existing drugs is the fastest and highly efficient approach to bring a therapeutic to a clinic for any new indication. One such drug that has been used extensively for decades is chloroquine (CQ, with its derivatives) either for malaria, lupus and rheumatoid arthritis. Accumulating body of evidence from experimental pharmacology suggests that CQ and related analogues also activate certain pathways that can potentially be exploited for therapeutic gain. For example, in the airways, this has opened an attractive avenue for developing novel bitter taste ligands as a new class of bronchodilators for asthma. While CQ and its derivatives have been proposed as a therapy in COVID-19, it remains to be seen whether it really work in the clinic? To this end, our perspective aims to provide a timely yet brief insights on the existing literature on CQ and the controversies surrounding its use in COVID-19. Further, we also highlight some of cell-based mechanism(s) that CQ and its derivatives affect in mediating variety of physiological responses in the cell. We believe, data emanating from the clinical studies and continual understanding of the fundamental mechanisms may potentially help in designing effective therapeutic strategies that meets both efficacy and safety criteria for COVID-19.

Doxorubicin (DOX) is an effective anthracycline used in chemotherapeutic regimens for a variety of haematological and solid tumors. However, its utility remains limited by its well-described, but poorly understood cardiotoxicity. Despite numerous studies describing various forms of regulated cell death and their involvement in DOX-mediated cardiotoxicity, the predominate form of cell death remains unclear. Part of this inconsistency lies in a lack of standardization of in vivo and in vitro model design. To this end, the objective of this study was to characterize acute low- and high-dose DOX exposure on cardiac structure and function in C57BL/6 N mice, and evaluate regulated cell death pathways and autophagy both in vivo and in cardiomyocyte culture models. Acute low-dose DOX had no significant impact on cardiac structure or function; however, acute high-dose DOX elicited substantial cardiac necrosis resulting in diminished cardiac mass and volume, with a corresponding reduced cardiac output, and without impacting ejection fraction or fibrosis. Low-dose DOX consistently activated caspase-signaling with evidence of mitochondrial permeability transition. However, acute high-dose DOX had only modest impact on common necrotic signaling pathways, but instead led to an inhibition in autophagic flux. Intriguingly, when autophagy was inhibited in cultured cardiomyoblasts, DOX-induced necrosis was enhanced. Collectively, these observations implicate inhibition of autophagy flux as an important component of the acute necrotic response to DOX, but also suggest that acute high-dose DOX exposure does not recapitulate the disease phenotype observed in human cardiotoxicity.

Glioblastoma (GBM) is human's most prevalent and severe brain cancer. Epigenetic regulators, micro(mi)RNAs, significantly impact cellular health and disease because of their wide range of targets and functions. The "epigenetic symphony" in which miRNAs perform is responsible for orchestrating the transcription of genetic information. The discovery of regulatory miRNA activities in GBM biology has shown that various miRNAs play a vital role in disease onset and development. Here, we summarize our current understanding of the current state-of-the-art and latest findings regarding the interactions between miRNAs and molecular mechanisms commonly associated with GBM pathogenesis. Moreover, by literature review and reconstruction of the GBM gene regulatory network, we uncovered the connection between miRNAs and critical signaling pathways such as cell proliferation, invasion, and cell death, which provides promising hints for identifying potential therapeutic targets for the treatment of GBM. In addition, the role of miRNAs in GBM patient survival was investigated. The present review, which contains new analyses of the previous literature, may lead to new avenues to explore in the future for the development of multitargeted miRNA-based therapies for GBM.

Abstract Current treatment strategies for Glioblastoma (GBM)—including surgery, radiotherapy, and chemotherapy with oral administration of temozolomide (TMZ)—still lead to poor survival rates, making the development of more effective therapeutic methods an urgent need. This study presents a new approach for the treatment of GBM patients using a 3D‐printed hydrogel‐based mesh (GlioMesh), loaded with TMZ‐releasing microparticles, that is capable of delivering TMZ over several weeks at the tumor site. Given the challenges associated with loading the amphiphilic TMZ in polymeric substrates, a novel encapsulation strategy is developed using an oil‐in‐oil emulsion method that improves the encapsulation efficiencies of TMZ in poly(lactic‐co‐glycolic acid) (PLGA) from &lt;7% to about 61%. The cytotoxic effects of GlioMesh on GBM cells are evaluated in vitro by investigating the resultant levels of DNA break, autophagic activity, and mitochondrial damage. It is shown that GlioMesh produces significantly higher susceptibility to the drug in comparison with free TMZ by maintaining the level of autophagic activity and inducing larger degrees of mitochondrial damage. Sustained delivery of TMZ holds promise for suppressing chemoresistance to TMZ that is normally developed in GBM cells in systemic administration of the drug due to the induction of autophagy.

Aim: MicroRNAs (miRNAs) are small noncoding RNAs that function as oncogene or tumor suppressors. The single nucleotide polymorphisms in miRNAs potentially can alter miRNA-binding sites on target genes as well as affecting miRNAs expression. The present study aimed to evaluate the impact of miR-608 rs4919510 C&gt;G variant on breast cancer (BC) risk. Materials and Me thods: This case-control study conducted on 160 women with BC and 192 age-matched healthy women. Genotyping of miR608 rs4919510 was done using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. Results: Our findings showed that GC genotype significantly decreased the risk of BC (odds ratio (OR) = 0.49, 95% confidence interval (CI) 0.28–0.88, p = 0.018) compared to CC genotype. Furthermore the G allele decreased the risk of BC (OR = 0.53, 95%CI 0.30–0.92, p = 0.024). No significant association was found between miR-609 genotypes and clinicopathological characteristics of BC patients (p &gt; 0.05). Conclusion: Our findings indicate that miR-608 polymorphism might be associated with decreased risk of BC in an Iranian subpopulation. Further large-scale studies with different ethnicities are needed to verify our findings.

Biomaterials for tissue scaffolds are key components in modern tissue engineering and regenerative medicine. Targeted reconstructive therapies require a proper choice of biomaterial and an adequate choice of cells to be seeded on it. The introduction of stem cells, and the transdifferentiation procedures, into regenerative medicine opened a new era and created new challenges for modern biomaterials. They must not only fulfill the mechanical functions of a scaffold for implanted cells and represent the expected mechanical strength of the artificial tissue, but furthermore, they should also assure their survival and, if possible, affect their desired way of differentiation. This paper aims to review how modern biomaterials, including synthetic (i.e., polylactic acid, polyurethane, polyvinyl alcohol, polyethylene terephthalate, ceramics) and natural (i.e., silk fibroin, decellularized scaffolds), both non-biodegradable and biodegradable, could influence (tissue) stem cells fate, regulate and direct their differentiation into desired target somatic cells.

Asthma is one of the fastest growing syndromes in many countries and is adding a huge cost to the health care system. Increasing reports have linked airway infectious diseases to asthma. Influenza is one of the most serious airway infectious diseases and in recent years there have been some serious influenza virus pandemics which caused increased fatality in numerous different populations. Diverse host response pathways during virus infection have been identified, including different cell death and survival pathways. These pathways include 1) programmed cell death I (apoptosis), 2) programmed cell death II (autophagy), and 3) endoplasmic reticulum stress with subsequent unfolded protein response (UPR). There has been extensive research on the regulatory roles of these pathways during the influenza virus life cycle. These studies address the benefits of enhancing or inhibiting these pathways on viral replication. Here we review the most recent and significant knowledge in this area for possible benefits to clinicians and basic scientist researchers in different areas of the respiratory and virology sciences.

Airway smooth muscle has classically been of interest for its contractile response linked to bronchoconstriction. However, terminally differentiated smooth muscle cells are phenotypically plastic and have multifunctional capacity for proliferation, cellular hypertrophy, migration, and the synthesis of extracellular matrix and inflammatory mediators. These latter properties of airway smooth muscle are important in airway remodeling which is a structural alteration that compounds the impact of contractile responses on limiting airway conductance. In this overview, we describe the important signaling components and the functional evidence supporting a view of smooth muscle cells at the core of fibroproliferative remodeling of hollow organs. Signal transduction components and events are summarized that control the basic cellular processes of proliferation, cell survival, apoptosis, and cellular migration. We delineate known intracellular control mechanisms and suggest future areas of interest to pursue to more fully understand factors that regulate normal myocyte function and airway remodeling in obstructive lung diseases.

Autoimmune hepatitis is a necroinflammatory process of liver, featuring interface hepatitis by T cells, macrophages and plasma cells that invade to periportal parenchyma. In this process, a variety of cytokines are secreted and liver tissues undergo fibrogenesis, resulting in the apoptosis of hepatocytes. Autophagy is a complementary mechanism for restraining intracellular pathogens to which the innate immune system does not provide efficient endocytosis. Hepatocytes with their particular regenerative features are normally in a quiescent state, and, autophagy controls the accumulation of excess products, therefore the liver serves as a basic model for the study of autophagy. Impairment of autophagy in the liver causes the accumulation of damaged organelles, misfolded proteins and exceeded lipids in hepatocytes as seen in metabolic diseases. In this review, we introduce autoimmune hepatitis in association with autophagy signaling. We also discuss some genes and proteins of autophagy, their regulatory roles in the activation of hepatic stellate cells and the importance of lipophagy and tyrosine kinase in hepatic fibrogenesis. In order to provide a comprehensive overview of the regulatory role of autophagy in autoimmune hepatitis, the pathway analysis of autophagy in autoimmune hepatitis is also included in this article.

Acute kidney injury (AKI) is an important complication of COVID-19 encompassing a wide range of presentations. SARS-CoV-2 is proposed to cause AKI in the patients through various mechanisms. We are, nevertheless, far from a comprehensive understanding of the underlying pathophysiological mechanisms of the kidney injury in this infection. AKI has been shown to be a marker of disease severity and also a negative prognostic factor for survival. Unfortunately, no effective preventive strategy to decrease the risk of kidney damage in these patients has yet been identified. In this hypothesis, we highlight the potential protective effects of acetazolamide, a carbonic anhydrase inhibitor, in preventing the proximal tubular damage caused by the virus through disrupting the virus-endosome fusion and also interfering with the lysosomal proteases. Our proposed mechanisms could pave the way for further in vitro studies and subsequent clinical trials.

Boswellia serrata gum is a natural product that showed beneficial effects on neurodegenerative diseases in recent studies. In this study, we investigated the effects of Boswellia serrata resin on rotenone-induced dopaminergic neurotoxicity. Firstly, we attempted to see if the resin can induce AMP-activated protein kinase (AMPK) signaling pathway which has been known to have broad neuroprotective effects. Boswellia increased AMPK phosphorylation and reduced phosphorylation of mammalian target of rapamycin (p-mTOR) and α-synuclein (p-α-synuclein) in the striatum while increased the expression level of Beclin1, a marker for autophagy and brain-derived neurotrophic factor. Next, we examined the neuroprotective effects of the Boswellia extract in the rotenone-injected mice. The results showed that Boswellia evidently attenuated the loss of the nigrostriatal dopaminergic neurons and microglial activation caused by rotenone. Moreover, Boswellia ameliorated rotenone-induced decrease in the striatal dopamine and impairment in motor function. Accumulation of α-synuclein meditated by rotenone was significantly ameliorated by Boswellia. Also, we showed that β-boswellic acid, the active constituents of Boswellia serrata gum, induced AMPK phosphorylation and attenuated α-synuclein phosphorylation in SHSY5 cells. These results suggest that Boswellia protected the dopaminergic neurons from rotenone neurotoxicity via activation of the AMPK pathway which might be associated with attenuation of α-synuclein aggregation and neuroinflammation. Further investigations are warranted to identify specific molecules in Boswellia which are responsible for the neuroprotection.

Background: The rapid spreading of corona virus disease 2019 (COVID-19) worldwide results in pneumonia and acute respiratory distress syndrome in many patients, which can be the major cause of death in cases with COVID-19. It has been reported that chloroquine (CQ) has improved COVID-19-induced pneumonia in clinical trials. Objectives: Since CQ and its derivatives are proved to exhibit anti-autophagy properties based on previous studies, autophagy can be introduced as a possible mechanism of respiratory complications. Methods: In the current study, we reviewed papers of Google Scholar database with no time limitation. Results: It was revealed that autophagy has an important role in the manifestation of COVID-19 respiratory complications Conclusions: Autophagy is triggered by SARS-CoV2 virus for its replication and autophagy inhibitory treatments might be considered promising therapeutics.

Clinical and Translational ScienceVolume 14, Issue 2 p. 431-433 CommentaryOpen Access Rationale for Effective Prophylaxis Against COVID-19 Through Simultaneous Blockade of Both Endosomal and Non-Endosomal SARS-CoV-2 Entry into Host Cell Seyed Fazel Nabavi, Seyed Fazel Nabavi Applied Biotechnology Research Center, Baqiyatallah University of Medical Sciences, Tehran, IranSearch for more papers by this authorSolomon Habtemariam, Solomon Habtemariam Pharmacognosy Research Laboratories and Herbal Analysis Services UK, University of Greenwich, Kent, UKSearch for more papers by this authorIoana Berindan-Neagoe, Ioana Berindan-Neagoe Department of Functional Genomics and Experimental Pathology, The Oncology Institute "Prof.Dr. Ion Chiricuta", Cluj-Napoca, RomaniaSearch for more papers by this authorCosmin Andrei Cismaru, Cosmin Andrei Cismaru Department of Functional Genomics and Experimental Pathology, The Oncology Institute "Prof.Dr. Ion Chiricuta", Cluj-Napoca, RomaniaSearch for more papers by this authorDedmer Schaafsma, Dedmer Schaafsma Science Impact, Winnipeg, Manitoba, CanadaSearch for more papers by this authorSaeid Ghavami, Saeid Ghavami Department of Human Anatomy and Cell Sciences, Rady Faculty of Health Sciences, Max Rady College of Medicine, University of Manitoba, Winnipeg, Manitoba, CanadaSearch for more papers by this authorMaciej Banach, Maciej Banach Department of Hypertension, Lodz, Medical University, Lodz, Poland Polish Mother's Memorial Hospital Research Institute (PMMHRI), Lodz, PolandSearch for more papers by this authorSafieh Aghaabdollahian, Safieh Aghaabdollahian Department of Nanobiotechnology, New Technologies Research Group, Pasteur Institute of Iran, Tehran, IranSearch for more papers by this authorSeyed Mohammad Nabavi, Corresponding Author Seyed Mohammad Nabavi [email protected] Applied Biotechnology Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran Correspondence: Seyed Mohammad Nabavi ([email protected])Search for more papers by this author Seyed Fazel Nabavi, Seyed Fazel Nabavi Applied Biotechnology Research Center, Baqiyatallah University of Medical Sciences, Tehran, IranSearch for more papers by this authorSolomon Habtemariam, Solomon Habtemariam Pharmacognosy Research Laboratories and Herbal Analysis Services UK, University of Greenwich, Kent, UKSearch for more papers by this authorIoana Berindan-Neagoe, Ioana Berindan-Neagoe Department of Functional Genomics and Experimental Pathology, The Oncology Institute "Prof.Dr. Ion Chiricuta", Cluj-Napoca, RomaniaSearch for more papers by this authorCosmin Andrei Cismaru, Cosmin Andrei Cismaru Department of Functional Genomics and Experimental Pathology, The Oncology Institute "Prof.Dr. Ion Chiricuta", Cluj-Napoca, RomaniaSearch for more papers by this authorDedmer Schaafsma, Dedmer Schaafsma Science Impact, Winnipeg, Manitoba, CanadaSearch for more papers by this authorSaeid Ghavami, Saeid Ghavami Department of Human Anatomy and Cell Sciences, Rady Faculty of Health Sciences, Max Rady College of Medicine, University of Manitoba, Winnipeg, Manitoba, CanadaSearch for more papers by this authorMaciej Banach, Maciej Banach Department of Hypertension, Lodz, Medical University, Lodz, Poland Polish Mother's Memorial Hospital Research Institute (PMMHRI), Lodz, PolandSearch for more papers by this authorSafieh Aghaabdollahian, Safieh Aghaabdollahian Department of Nanobiotechnology, New Technologies Research Group, Pasteur Institute of Iran, Tehran, IranSearch for more papers by this authorSeyed Mohammad Nabavi, Corresponding Author Seyed Mohammad Nabavi [email protected] Applied Biotechnology Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran Correspondence: Seyed Mohammad Nabavi ([email protected])Search for more papers by this author First published: 06 January 2021 https://doi.org/10.1111/cts.12949Citations: 5AboutSectionsPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL The severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) enters host cells through the endosomal and nonendosomal pathways. Although the relative contribution of each mechanism could vary depending on host cell types, evidence from studies on related coronaviruses suggest that simultaneous targeting of both pathways could be an efficient therapeutic approach to limit viral infection. This communication highlights the prophylactic potential of combination therapies using TMPRSS2, cathepsin/calpain, and FURIN inhibitors for complete blockade of SARS-CoV-2 infection. Until 2 decades ago, several human coronaviruses were acknowledged, but were given little attention as they only caused mild upper respiratory tract infections with a severity comparable to common cold virus infections. However, the emergence of the severe acute respiratory syndrome (SARS) in China (2002/2003) and Middle East respiratory syndrome (MERS) in Saudi Arabia (2012) changed the overall dynamics of our focus on coronaviruses, both in vaccine development and therapeutics. The current global medical and socioeconomic cost associated with SARS-CoV-2 (the newly discovered coronavirus) infection, first reported in the Wuhan region of China in December 2019, is of a magnitude exceeding any other over the last generations. With over 21 million cases reported so far, the ongoing SARS-CoV-2 pandemic, referred to as coronavirus disease-2019 (COVID-19), has already claimed over 1 million human lives across the globe. Regarding therapeutics for COVID-19, the 2 main approaches are based on targeting (1) the key viral proteins/enzymes involved in the replication cycle of SARS-CoV-2, or (2) the host immune system by enhancing clearance of the virus and/or suppressing the overexaggerated immune reaction leading to multiorgan failure and death. In this paper, the therapeutic potential of targeting the entry mechanisms of the coronavirus is discussed. Coronaviruses can use two mechanisms of entry into host cells: the endosomal and nonendosomal pathways (Figure 1).1 In view of interrupting the SARS-CoV-2 infection cycle by targeting the nonendosomal pathway, the key steps in processing the viral spike (S) proteins, their attachment with host receptors, in particular angiotensin converting enzyme 2 (ACE2), and the viral entry mechanisms have been scrutinized since the outbreak of COVID-19. Hoffmann et al. highlighted the role of TMPRSS2 priming of the S glycoprotein as part of the nonendosomal SARS-CoV-2 cellular entry pathway and the potential application of its inhibitor camostat mesylate as a potential therapeutic agent.2 Other approaches include using neutralizing antibodies against TMPRSS2.2 Based on these and other findings, the proteolytic cleavage of inactive S proteins, which is required to generate an active viral ligand suitable for attachment to ACE2, has emerged as a key target of interest.2 Figure 1Open in figure viewerPowerPoint Endosomal and nonendosomal severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) entry into host cells. The endosomal mechanisms of coronavirus entry into host cells have been well-established and require the activity of the pH-dependent cysteine proteases cathepsin B and L.1 Although further research is required to validate their therapeutic efficacy and overcome certain safety concerns, the claimed benefit of chloroquine and hydroxychloroquine for COVID-19 is based on targeting the endosomal pathway (pH) of SARS-CoV-2 entry. Although these pharmacological mechanisms are proven under in vitro infection models, readers should bear in mind that these antimalarial drugs have multiple mechanisms and their efficacy against COVID-19 is not yet to be confirmed, as some clinical trials have already been discontinued for various reasons. SARS-CoV replication was suggested to be significantly higher following entry through the nonendosomal than the endosomal pathway.3 It is, however, rational to assume that targeting both entry mechanisms simultaneously would offer a superior outcome in effectively disrupting the SARS-CoV infection cycle. Supporting this notion, combined treatment with camostat (TMPRSS2 inhibitor) and EST (cathepsin L and/or B inhibitor) effectively prevented cell entry and multistep growth of SARS-CoV in human airway epithelial cells in vitro, which could be attributed to dual inhibition of the endosomal pathway and entry from the cell surface.4 The combination of camostat and a cathepsin inhibitor ((23,25)-trans-epoxysuccinyl-l-leucylamindo-3-methylbutane ethyl ester) have been studied in relation to MERS-CoV as well.5 The results revealed that simultaneous inhibition of TMPRSS2 and cathepsin L completely blocked MERS-CoV entry into Vero-TMPRSS2 cells, confirming that, as with for SARS-CoV, this virus uses both the cell surface and endosomal pathway to infect the host. In addition, camostat was sufficient to block MERS-CoV entry in human bronchial submucosal gland-derived Calu-3 cells, as the combined treatment with the cathepsin inhibitor was no more efficacious than treatment with camostat alone. This latter finding might indicate differential contributions of the cell surface and endosome pathway to viral entry depending on the cell type. These studies further imply that agents inhibiting host serine and/or cysteine proteases, relevant to viral cell entry, could effectively interfere with coronavirus infection. In addition to playing a key role in the endosomal pathway, cathepsin L proteolysis has been shown to contribute to TMPRSS2-mediated S protein activation and membrane fusion, linking it to cell surface-mediated SARS-CoV entry into the host cells.6 All these promising data based on MERS-CoV and SARS-CoV are significant milestones but yet to be translated to SARS-CoV-2 therapy through further studies. Pretreatment of 293/hACE2 cells with E64D (a broad inhibitor for cathepsin B, H, L, and calpain) strongly reduced entry of SARS-CoV-2 S pseudovirions (by 92.5%), suggesting the requirement of at least one of the cathepsins or calpain. Further studies using specific inhibitors demonstrated over 76% reduction in cell entry when a cathepsin L (SID-26681509), but not B inhibitor (CA-074), was used, implying that particularly cathepsin L is critical for priming the SARS-CoV-2 S protein in the lysosome for entry into these cells.7 Recent study showed simultaneous treatment with camostat mesylate and E64d markedly inhibited SARS-CoV-2 entry into Calu-3 cells, but could not completely abrogate viral entry.8 The S protein of SARS-CoV-2 contains a PRRA motif, which contains four redundant FURIN cleavage sites.8 Notably, a FURIN-like cleavage site in the S-protein of the SARS-CoV-2 was identified as unique, as it was lacking in the other SARS-like coronaviruses.8 Hoffmann et al.2 speculated that FURIN may be involved in SARS-CoV-2 S protein priming. Luteolin is a well-known, safe, and potent inhibitor of FURIN with a inhibitor constant value of 58.6 μM, and has been shown to exhibit antiviral activity in mice infected with dengue virus.9 This small molecule has also potent antiviral effect against wild-type SARS-CoV infection in Vero E6 cells (half-maximal effective concentration: 10.6 µm).10 Overall, the data discussed in this short communication suggest that simultaneous targeting of endosomal and nonendosomal cellular entry of SARS-CoV-2 could represent a highly efficient therapeutic approach to interfere with viral infection. In this context, we feel it would be valuable to test the prophylactic potential of combination therapies of TMPRSS2 inhibitors (such as ambroxol, leupeptin, camostat mesylate, and nafamostat mesylate), Aloxistatin (E64d) (as potent inhibitor of cathepsin L and calpain) in association with FURIN inhibitors (e.g., luteolin) that may lead to complete abrogation of SARS-CoV-2 entry into host cells. Funding No funding was received for this work. Conflict of Interest The authors declared no competing interests for this work. References 1Zumla, A., Chan, J.F., Azhar, E.I., Hui, D.S. & Yuen, K.Y. Coronaviruses—drug discovery and therapeutic options. Nat. Rev. Drug Discov. 15, 327– 347 (2016). 2Hoffmann, M. et al. SARS-CoV-2 cell entry depends on ACE2 and TMPRSS2 and is blocked by a clinically proven protease inhibitor. Cell 181, 271– 280.e8 (2020). 3Lukassen, S. et al. SARS-CoV-2 receptor ACE 2 and TMPRSS 2 are primarily expressed in bronchial transient secretory cells. EMBO J. 39, e105114 (2020). 4Kawase, M., Shirato, K., van der Hoek, L., Taguchi, F. & Matsuyama, S. Simultaneous treatment of human bronchial epithelial cells with serine and cysteine protease inhibitors prevents severe acute respiratory syndrome coronavirus entry. J Virol. 86, 6537– 6545 (2012). 5Shirato, K., Kawase, M. & Matsuyama, S. Middle East respiratory syndrome coronavirus infection mediated by the transmembrane serine protease TMPRSS2. J. Virol. 87, 12552– 12561 (2013). 6Simmons, G., Gosalia, D.N., Rennekamp, A.J., Reeves, J.D., Diamond, S.L. & Bates, P. Inhibitors of cathepsin L prevent severe acute respiratory syndrome coronavirus entry. Proc. Natl. Acad. Sci. 102, 11876– 11881 (2005). 7Ou, X. et al. Characterization of spike glycoprotein of SARS-CoV-2 on virus entry and its immune cross-reactivity with SARS-CoV. Nat. Commun. 11, 1620 (2020). 8Mykytyn, A.Z. et al. The SARS-CoV-2 multibasic cleavage site facilitates early serine protease-mediated entry into organoid-derived human airway cells. bioRxiv. https://doi.org/10.1101/2020.09.07.286120. 9Peng, M. et al. Luteolin restricts dengue virus replication through inhibition of the proprotein convertase FURIN. Antiviral Res. 143, 176– 185 (2017). 10Yi, L. et al. Small molecules blocking the entry of severe acute respiratory syndrome coronavirus into host cells. J. Virol. 78, 11334– 11339 (2004). Citing Literature Volume14, Issue2March 2021Pages 431-433 This article also appears in:COVID-19 in ASCPT FiguresReferencesRelatedInformation

Epithelial-mesenchymal transition (EMT) and the stemness potency in association with BRAF mutation are in dispensable to the progression of melanoma. Recently, microRNAs (miRNAs) have been introduced as the regulator of a multitude of oncogenic functions in most of tumors. Therefore identifying and interpreting the expression patterns of these miRNAs is essential. The present study sought to find common miRNAs regulating all three important pathways in melanoma development.In this experimental study, 18 miRNAs that importantly contribute to EMT and have a role in regulating self-renewal and the BRAF pathway were selected based on current literature and cross-analysis with available databases. Subsequently, their expression patterns were evaluated in 20 melanoma patients, normal tissues, serum from patients and control subjects, and melanospheres. Pattern discovery and integrative regulatory network analysis were used to find the most important miRNAs in melanoma progression.Among 18 selected miRNAs, miR-205, -141, -203, -15b, and -9 were differentially expressed in tumor samples than normal tissues. Among them, miR-205, -15b, and -9 significantly expressed in serum samples and healthy donors. Attribute Weighting and decision trees (DT) analysis presented evidence that the combination of miR-205, -203, -9, and -15b can regulate self-renewal and EMT process, by affecting CDH1, CCND1, and VEGF expression.We suggested here that miR-205, -15b, -203, -9 pattern as the key miRNAs linked to melanoma status, the pluripotency, proliferation, and motility of malignant cells. However, further investigations are required to find the mechanisms underlying the combinatory effects of the above mentioned miRNAs.

Nanotechnology is a fast-growing field with the broad application of magnetic nanoparticles (MNPs) in different facets of modern human life being a perfect example. MNPs have recently emerged in many industrial technologies and devices related to biomedical, environmental, and clinical disciplines. Such a variety of critical applications of MNPs triggered new important areas of research; this chapter is primarily focused on three main themes: (1) basic characteristics of long-term interaction/behavior between MNPs and biological life, (2) pros and cons of their toxic effects on human cells, and (3) state-of-the-art techniques for safe disposal/recycling of consumed MNPs. Additionally, we highlight the importance of some toxic characteristics of MNPs in cancer treatment through hyperthermia phenomena. We also discussed the importance of MNPs in targeting cancer cells and their application in modern chemotherapy strategies.

Macrophages and T-lymphocytes are involved in immune response to Mycobacterium tuberculosis. Macrophage produces interleukin (IL)-1 as an inflammatory mediator. IL-1 receptor antagonist (IL1-Ra) is a natural antagonist of IL-1 receptors. In this study we aimed to examine the possible association between the variable number of tandem repeats (VNTR) of the IL-1 receptor antagonist (IL1RN) gene and pulmonary tuberculosis (TB) in a sample of Iranian population. Our study is a case-control study and we examined the VNTR of the IL1RN gene in 265 PTB and 250 healthy subjects by PCR. Neither the overall chi-square comparison of PTB and control subjects nor the logistic regression analysis indicated any association between VNTR IL1RN polymorphism and PTB. Our data suggest that VNTR IL1RN polymorphism may not be associated with the risk of PTB in a sample of Iranian population. Larger studies with different ethnicities are needed to find out the impact of IL1RN VNTR polymorphism on risk of developing TB.

Rhabdomyosarcoma (RMS) is a muscle-derived tumor and is the most common pediatric soft tissue sarcoma representing 5% of all childhood cancers.Statistically, soft tissue sarcomas account for approximately 10% of all cancers in children, of which more than half of these tumors are RMS.Thus, RMS is a major clinical problem in pediatric oncology.RMS is caused by a disruption in the pathway of primitive mesenchymal stem cells directed towards myogenesis.In most cases of patients diagnosed with RMS there is a genetic or chromosomal alteration involved.In past few years there have been discoveries of more therapeutic approaches that has improved the quality of life in RMS patients and has resulted in a better survival rate in this population from 25% to 60%.However, Additional researches and clinical trials are needed in order to minimize the devastating consequences of the pediatric cancer including RMS.In the current mini review we will briefly discuss current knowledge in RMS focusing on most common biological and clinical aspects of the disease.

Inflammatory bowel diseases (IBDs) are chronic disorders of the gastrointestinal tract. The goal of IBD treatment is to reduce the inflammation period and induce long-term remission. Use of anti-inflammatory drugs including corticosteroids, immunosuppressants and biologicals, is often the first step in the treatment of IBD. Therefore, IBD patients in pandemic of infectious diseases are considered a high-risk group. The public believes that IBD patients are at a higher risk in the current coronavirus 2 pandemic. Nevertheless, these patients may experience mild or moderate complications compared to healthy people. This might be because of particular anti-TNF-α treatment or any immunosuppressant that IBD patients receive. Moreover, these patients might be silent carrier for the virus.

Introduction: Lymphoma is a common hematopoietic cancer. Immunosuppression is one of the main risk factors for the development of lymphoma. The interleukin (IL)-1 receptor antagonist IL1RN, which binds to the IL-1 receptor, moderates a variety of immune responses related to IL-1. We aimed to assess the impact of IL1RN variable number of tandem repeats (VNTR) polymorphism on lymphoma risk in an Iranian population sample.&#x0D; Materials and Methods: DNA was extracted from peripheral blood of 120 subjects with non-Hodgkin Lymphoma (NHL), 50 subjects with Hodgkin’s lymphoma (HL), and 186 unrelated healthy individuals. IL1RN VNTR polymorphism was detected using polymerase chain reaction.&#x0D; Results: Our findings revealed that the IL1RN VNTR polymorphism was associated with protection against NHL (P≤0.001, OR: 0.30, 95% CI: 0.18-0.53). The IL1RN 2 allele significantly decreased the risk of NHL (p = 0.023, OR = 0.66, 95%CI = 0.46–0.93). In addition, we found that IL1RN 1/2 was associated with a lower risk of HL (p ≤0.001, OR = 0.24, 95%CI = 0.12–0.50).&#x0D; Conclusion: Our results suggest that the presence of IL1RN VNTR polymorphism is associated with a decreased risk of lymphoma in an Iranian subpopulation in southeast Iran.

Accumulated evidence have proposed that single nucleotide polymorphisms (SNPs) in microRNAs (miRNAs) are connected to breast cancer (BC) risk. We have done a case-control study with 258 BC patients and 209 control women to examine the potential association of Hsa-mir-603 rs11014002 C>T polymorphisms with BC susceptibility. The polymorphisms were genotyped by polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) method. Our findings showed that the rs11014002 C>T variant was not associated with an increased risk of BC in codominant (OR=0.67, 95%CI=0.42-1.08, P=0.121, CT vs CC; and OR=0.18, 95%CI=0.02-1.67, P=0.170, TT vs CC), dominant (OR=0.64, 95%CI=0.41-1.01, P=0.062, CT+TT vs CC), and recessive (OR=0.20, 95%CI=0.02-1.81, P=0.178, TT vs CC+CT) inheritance models tested. While, the T allele significantly decreased the risk of BC (OR= 0.63; 95% CI =0.41-0.95; P=0.032) compared to C allele. In conclusion, the findings indicated that Mir603 rs11014002 T allele might contribute to decrease the risk of BC in a sample of Iranian population. Further studies with larger sample sizes and different ethnicities are warranted to confirm our findings.

Genetic polymorphisms in ERBB4 are thought to be associated with cancer susceptibility. In the present study, we aimed to assess the impact of ERBB4 rs12052398 T>C, rs13393577 A>G, rs13424871 A>T, rs16847082 A>G and rs6147150 (12-bp I/D) polymorphisms on risk of prostate cancer (PCa) in a sample of Iranian population. In a case-control study, we enrolled 169 patients with pathologically confirmed PCa and 182 subjects with benign prostatic hyperplasia (BPH). No significant association was found among ERBB4 polymorphisms and risk of PCa. Subjects carrying TT/AA/AA/AG/ID, TC/AA/AA/AA/II, TT/AA/AT/AA/II and TT/AA/AT/AG/ID genotypes are associated with a decreased risk of PCa. Our findings suggest that haplotypes CAAAI and TAAAD (rs12052398, rs13393577, rs13424871, rs16847082 and rs6147150I) of the ERBB4 polymorphisms are associated with a significantly lower risk of PCa. Further studies with a larger sample sizes and diverse ethnicities are necessary to verify our findings.

Purkinje cells (PCs) are large GABAergic projection neurons of the cerebellar cortex, endowed with elaborate dendrites that receive a multitude of excitatory inputs. Being the only efferent neuron of the cerebellar cortex, PCs project to cerebellar nuclei and control behaviors ranging from movement to cognition and social interaction. Neural cell adhesion molecule 1 (NCAM1) is widely expressed in the embryonic and postnatal development of the brain and plays essential roles in neuronal migration, axon pathfinding and synapse assembly. However, despite its high expression levels in cerebellum, little is known to date regarding the role(s) of NCAM1 in PCs development. Among other aspects, elucidating how the expression of NCAM1 in PCs could impact their postnatal migration would be a significant achievement. We analyzed the Acp2 mutant mouse (nax: naked and ataxia), which displays excessive PC migration into the molecular layer, and investigated how the excessive migration of PCs along Bergmann glia could correlate to NCAM1 expression pattern in early postnatal days. Our Western blot and RT-qPCR analysis of the whole cerebellum show that the protein and mRNA of NCAM1 in wild type are not different during PC dispersal from the cluster stage to monolayer formation. However, RT-qPCR analysis from FACS-based isolated PCs shows that Ncam1 is significantly upregulated when PCs fail to align and instead overmigrate into the molecular layer. Our results suggest two alternative interpretations: (1) NCAM1 promotes excessive PC migration along Bergmann glia, or (2) NCAM1 upregulation is an attempt to prevent PCs from invading the molecular layer. If the latter scenario proves true, NCAM1 may play a key role in PC monolayer formation.

It has been established that overproduction of reactive oxygen species (ROS) occurs during hemodialysis causing oxidation of proteins. Alpha-1-antitrypsin is the major circulating anti-protease which contains methionine in the active site. The aim of the present study was to measure the level of serum trypsin inhibitory capacity (sTIC) in hemodialysis patients. This case-control study was performed in 52 hemodialysis patients and 49 healthy controls. sTIC was measured by enzymatic assay. The sTIC was significantly (P< 0.001) lower in hemodialysis patients (1.87 +/- 0.67 micromol/min/mL) than healthy controls (2.83 +/- 0.44 micromol/min/L). Reduction of sTIC may be due to the oxidation of methionine residue in the reactive site of alpha-1 antitrypsin.

Background: Interleukin 27 (IL-27) has potent antitumor activity. Several epidemiological studies have designated that genetic variants of the IL-27 gene may contribute to various cancer susceptibility, but the data were inconclusive. Objective: The current meta-analysis aimed to address the association between IL-27 rs153109, rs17855750, and rs181206 polymorphisms and the risk of cancer. Data Sources: Our team has selected eligible studies up to May 1, 2020, from several electronic databases, including Web of Science, PubMed, Scopus, and Google Scholar databases. Results: Our meta-analysis revealed that the carriers rs153109 A>G polymorphism in the IL-27 gene have higher risks of diseases in the heterozygous (OR=1.26, 95%CI=1.06-1.49, P=0.007, AG vs AA), homozygous (OR=1.18, 95%CI=1.01-1.37, p=0.33, GG vs AA), dominant (OR=1.25, 95%CI=1.07-1.47, P=0.006, AG+GG vs AA), and allele (OR=1.15, 95%CI=1.04-1.27, P=0.008, G vs A) genetic models. Stratified analysis by cancer type indicated that this variant was significantly associated with gastrointestinal cancer, colorectal cancer and breast cancer. The findings did not support an association between rs17855750 T>G, rs181206 T>C polymorphisms of IL-27 and cancer risk. Conclusion: the current study findings suggest that IL-27 rs153109 polymorphism significantly increased the risk of cancer susceptibility. Well-designed replication in a larger independent genetic association study with larger sample sizes in diverse ethnicities is required to verify the findings.

Background: The AKT/PKB (protein kinase B) kinase is the main regulator of autophagy in mammalian cells, which consists of three isoforms, including AKT-1, AKT-2, and AKT-3. Rat sarcoma viral oncogene homolog (RAS), known as the most frequently mutated oncogene in colorectal cancers, is one of the major activators of AKT signaling. However, the relationship between AKT isoforms expression and autophagy level in RAS-driven cancer cells has not been fully investigated. Method: In this experimental in vitro study, RAS mutated colon cancer cell lines (HCT116, SW480, and LS180) and HT29 cells, which are the wild type of RAS, were cultured and real-time polymerase chain reaction (RT-PCR) was utilized to determine the mRNA level of AKT-1, AKT-2, and autophagy markers, including microtubule-associated protein 1 light chain-3B (LC3B) and p62/sequestosome-1 (p62). In addition, Western blotting was performed to assess the protein expression of p62 and LC3B lipidation. Results: We found that RAS mutated colon cancer cells up-regulate basal autophagy. Moreover, highly expressed AKT-1 was observed in RAS mutated colon cancer cells. However, no significant differences were found in AKT-2 expression between RAS-driven cells and HT29 cells. Conclusion: Our obtained data suggested that RAS-driven colon cancer cells regulated the autophagy machinery, possibly, through the upregulation of AKT-1 isoform.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), which causes novel coronavirus disease (COVID-19), is the seventh pathogenic coronavirus recently discovered in December 2019 in Wuhan, China. To date, our knowledge about its effect on the human host remains limited. It is well known that host genetic factors account for the individual differences in the susceptibility to infectious diseases. The genetic susceptibility factors to COVID-19 and its severity are associated with several unanswered questions. However, the experience gained from an earlier strain of coronavirus, SARS-CoV-1, which shows 78% genetic similarity to SARS-CoV-2 and uses the same receptor to bind to host cells, could provide some clues. It, therefore, seems possible to assemble new evidence in order to solve a potential genetic predisposition puzzle for COVID-19. In this chapter, the puzzle pieces, including virus entry receptors, immune response, and inflammation-related genes, as well as the probable genetic predisposition models to COVID-19, are discussed.

Glioblastoma (GBM) is a rapidly progressive form of brain tumor with high mortality rates. Current treatment modalities are moderately effective and chemoresistance has become a major concern in GBM treatment. Evidence suggests that cross talks between several pathways, including apoptosis, autophagy, and unfolded protein response (UPR), are involved in the induction of resistance to chemotherapeutic agents such as temozolomide (TMZ). Apoptosis is crucial for organism survival through the elimination of unwanted cells. Various conditions such as hypoxia, defective DNA mismatch repair (MMR) system, p53 loss of function, insufficient O6-methylguanine-DNA-methyltransferase (MGMT) enzyme, and dysregulation of miRNAs expression negatively influence apoptosis and lead to chemoresistance. A dual role has been reported for autophagy in GBM chemoresistance and chemosensitivity. Similarly, the UPR adaptive pathway has both cytotoxic and cytoprotective effects on GBM cells in response to chemotherapeutic agents. Considering the complexity of this malignancy, the development of comprehensive in vitro models is required to predict the effects of different therapeutic strategies by recapitulating the intricate in vivo GBM microenvironment. Three-dimensional (3D) tissue engineering using different biomaterials is a promising approach to evaluate the effectiveness of chemotherapeutics by developing bioengineered GBM models. The functional contribution of signaling pathways such as autophagy, apoptosis, and UPR can be investigated in detail through 3D tissue engineering. In this chapter, we briefly review the role of autophagy, apoptosis, and unfolded protein response in chemoresistance in GBM and later discuss the application of a 3D bioengineer model of GBM and its application in the evaluation of chemotherapy response and chemoresistance in GBM.

Review on S100A9 (S100 calcium binding protein A9), with data on DNA, on the protein encoded, and where the gene is implicated.

Development is an evolutionary process that is tightly regulated in mammalian species. Several different cascades are involved in different stages of development. Among these mechanisms, apoptosis, autophagy, and unfolded protein response play critical roles in regulation of development by affecting the cell fate. All of these pathways are involved in regulation of cell number via determining the life and death cycles of the cells. In this chapter, we first explain the brief mechanisms that are involved in regulation of apoptosis, autophagy, and unfolded protein response, and later, we briefly describe how these mechanisms play roles in general development. We then discuss the importance of these pathways in regulation of cerebellar development.

The activation of hepatic stellate cells is the primary function of facilitating liver fibrosis. Interfering with the coordinators of different signaling pathways in activated hepatic stellate cells (aHSCs) could be a potential approach in ameliorating liver fibrosis. Regarding the illustrated anti-fibrotic effect of imatinib in liver fibrosis, we investigated the imatinib's potential role in inhibiting HSC activation through miR-124 and its interference with the STAT3/hepatic leukemia factor (HLF)/IL-6 circuit. The anti-fibrotic effect of imatinib was investigated in the LX-2 cell line and carbon tetrachloride (CCl4 )-induced Sprague-Dawley rat. The expression of IL-6, STAT3, HLF, miR-124, and α-smooth muscle actin (α-SMA) were quantified by quantitative real-time PCR (qRT-PCR) and the protein level of α-SMA and STAT3 was measured by western blot analysis both in vitro and in vivo. The LX-2 cells were subjected to immunocytochemistry (ICC) for α-SMA expression. After administering imatinib in the liver fibrosis model, histopathological examinations were done, and hepatic function serum markers were checked. Imatinib administration alleviated mentioned liver fibrosis markers. The expression of miR-124 was downregulated, while IL-6/HLF/STAT3 circuit agents were upregulated in vitro and in vivo. Notably, imatinib intervention decreased the expression of IL-6, STAT3, and HLF. Elevated expression of miR-124 suppressed the expression of STAT3 and further inhibited HSCs activation. Our results demonstrated that imatinib not only ameliorated hepatic fibrosis through tyrosine kinase inhibitor (TKI) activity but also interfered with the miR-124 and STAT3/HLF/IL-6 pathway. Considering the important role of miR-124 in regulating liver fibrosis and HSCs activation, imatinib may exert its anti-fibrotic activity through miR-124.

Development is an evolutionary process that is tightly regulated in mammalian species. Several different cascades are involved in various stages of development. Among these mechanisms, apoptosis, autophagy, and unfolded protein response play critical roles in regulating development by affecting cell fate. All of these pathways are involved in the regulation of cell numbers via determining the life and death cycles of the cells. In this chapter, we first explain the brief mechanisms that are involved in the regulation of apoptosis, autophagy, and unfolded protein response, and later, we briefly describe how these mechanisms play roles in general development. We next address the critical role of these pathways in cerebellar development regulation and how they will aid in our knowledge of the processes behind neurodevelopmental disorders. Additionally, we summarize the present findings on neurological symptoms and disorders related to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection and their linkage to autophagy pathways in the cerebellum.

Mitochondrial quality control is critical in muscle to ensure both contractile and metabolic function. Homeostatic pathways operate to ensure mitochondrial health by coordinating mitochondrial biogenesis with the removal of damaged mitochondrial components, which have been collectively termed mitochondrial quality control. In addition, mitochondria can respond to cell stress by integrating signaling pathways that orchestrate a tissue level response, but the mechanisms for this are largely unknown. Nix is a BCL-2 family member, mitophagy receptor, and has recently been implicated in muscle atrophy and aging. In human and rodent myotubes, we previously demonstrated that Nix orchestrates both mitochondrial calcium and mTOR signaling in responsive to a lipotoxic stress leading mitochondrial turnover and impaired insulin signaling. In addition, human GWAS suggests altered Nix expression could predispose individuals to manifestations of mitochondrial disease.

Mitochondrial quality control is critical in muscle to ensure both contractile and metabolic function. Homeostatic pathways operate to ensure mitochondrial health by coordinating mitochondrial biogenesis with the removal of damaged mitochondrial components, which have been collectively termed mitochondrial quality control. In addition, mitochondria can respond to cell stress by integrating signaling pathways that orchestrate a tissue level response, but the mechanisms for this are largely unknown. Nix is a BCL-2 family member, mitophagy receptor, and has recently been implicated in muscle atrophy and aging. In human and rodent myotubes, we previously demonstrated that Nix orchestrates both mitochondrial calcium and mTOR signaling in responsive to a lipotoxic stress leading mitochondrial turnover and impaired insulin signaling. In addition, human GWAS suggests altered Nix expression could predispose individuals to manifestations of mitochondrial disease.

Calprotectin, a calcium-bound protein complex, is abundant in the cytosol of neutrophils. It has been reported that this protein has an apoptotic activity in tumor cells. Since calprotectin increases in colorectal cancer, this study was conducted to investigate, for the first time, the cytotoxicity/apoptotic effect of calprotectin on HT29/219 and SW742 colon carcinoma and adenocarcinoma cell lines. MTT (3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide) assay, annexin V/PI and Hoechst 33258 staining were used to detect apoptotic cells. The activation of caspase-3 and -9 was assayed using caspase assay kits. Flow cytometer was used to determine if there is any binding of calprotectin to its receptor. Treatment of the cells with different concentrations of human calprotectin resulted in a significant increase in the cell death. Annexin V/PI and Hoechst 33258 staining revealed that the cell death was mainly of apoptotic type. A significant (p<0.05) increase in the activity of caspase-3, and -9 was observed in both cell lines following the treatment. However no binding activity was observed. In Conclusion, this study showed that human calprotectin has an apoptotic effect on HT29/219 and SW742 cells, and that the mode of action is caspase activation particularly via mitochondrial route.

Abstract Lipotoxicity is a form of cellular stress caused by the accumulation of lipids resulting in mitochondrial dysfunction and insulin resistance in muscle. Previously, we demonstrated that the mitophagy receptor Nix is responsive to lipotoxicity and accumulates in response to diacylglycerols induced by high-fat (HF) feeding. In addition, previous studies have implicated autophagy and mitophagy in muscle insulin sensitivity. To provide a better understanding of these observations, we undertook gene expression array and shot-gun metabolomics studies in soleus muscle from rodents on an HF diet. Interestingly, we observed a modest reduction in several autophagy-related genes including Beclin-1, ATG3, and -5. Moreover, we observed alterations in the fatty acyl composition of cardiolipins and phosphatidic acids. Given the previously reported roles of these phospholipids and Nix in mitochondrial dynamics, we investigated aberrant mitochondrial fission and turn-over as a mechanism of myocyte insulin resistance. In a series of gain-of-function and loss-of-function experiments in rodent and human myotubes, we demonstrate that Nix accumulation triggers mitochondrial depolarization, fragmentation, calcium-dependent activation of DRP1, and mitophagy. In addition, Nix-induced mitochondrial fission leads to myotube insulin resistance through activation of mTOR-p70S6 kinase inhibition of IRS1, which is contingent on phosphatidic acids and Rheb. Finally, we demonstrate that Nix-induced mitophagy and insulin resistance can be reversed by direct phosphorylation of Nix by PKA, leading to the translocation of Nix from the mitochondria and sarcoplasmic reticulum to the cytosol. These findings provide insight into the role of Nix-induced mitophagy and myocyte insulin resistance during an overfed state when overall autophagy-related gene expression is reduced. Furthermore, our data suggests a mechanism by which exercise or pharmacological activation of PKA may overcome myocyte insulin resistance. Graphical Abstract

Unfolded protein response (UPR) pathway is a promising target for cancer treatment because of its over-activation in different cancers and its role in tumorigenesis and chemotherapeutic drug resistance. Endoplasmic Reticulum Metallo Protease 1 (ERMP1) is overexpressed in cancers such as colorectal cancer. The ERMP1 role in UPR activation was previously reported in breast cancer. We aimed to investigate the ERMP1 role in the UPR activation in colorectal cancer. In this regard, ERMP1 gene was silenced in colorectal cancer HCT116 cell line using specific small hairpin RNA (shRNA). Then, UPR associated protein markers including inositol requiring enzyme 1 (IRE1α), activating transcription factor 6 (ATF-6), eukaryotic initiation factor 2α (eIF2α) and phosphorylated eIF2α (P- eIF2α) were evaluated using western blot. We found that ERMP1 gene expression and all of the above UPR associated protein markers were significantly decreased after ERMP1 gene silencing. Therefore, it seems that ERMP1 plays an important role in UPR activation. Since the overexpression of ERMP1 as a potential oncogene can highly activate the UPR pathway in colorectal cancer, it can be considered as a promising target for colorectal cancer treatment. However, further investigations are required to confirm these findings.

We investigate the impact of IL-1A, IL-1B and IL-1R1 polymorphism on lymphoma. This study consisted of 155 Non-Hodgkin’s lymphoma (NHL) patients 55 Hodgkin’s lymphoma (HL) patients and 150 healthy individuals. PCR-RFLP method and ARMS PCR were used for genotyping of IL-1A rs3783553, IL-1B rs3917356, rs16944, IL-1R1 rs10490571 and IL-1A rs3783550 polymorphism. The results showed that the CC genotype of rs3783550 as well Ins/del of rs3783553 increased the risk of NHL. In contrast the AG genotype of rs3917356 and AG also AG + AA genotype of rs10490571 decreased the risk of NHL. The result revealed that the CC genotype of rs3783550 and AG genotype of rs3917356 increased risk of HL.

Glioblastoma multiforme (GBM) is the deadliest brain tumor with an approximate 20 month survival rate after diagnosis and treatment. Temozolomide (TMZ), the chemotherapeutic drug of choice for GBM, is an alkylating agent that causes DNA damage. TMZ treatment results in the induction of apoptosis in GBM cells, however, it induces autophagy and consequently chemoresistance. Statins are mevalonate (MEV) cascade inhibitors with beneficial effects on the enhancement of the survival rate of patients with different types of cancer, particulary if consumed long term before diagnosis of cancer. Here, we determined the effect of simvastatin (Simva), a blood brain barrier permeable statin, on the sensitization of GBM cells to TMZ induced apoptosis through inhibition of autophagy flux. We pretreated two GMB cell lines, U251 and U87 cells, with low doses of Simva (1 and 2.5 μM, respectively) with or without different intermediates of the mevalonate cascade: MEV (2.5 mM), Farnesyl pyrophosphate (FPP) (15 μM), Geranylgeranyl pyrophosphate (GGPP) (15 μM) and cholesterol (50 μM) for 4 h and then treated cells with TMZ (100 μM) for 48–96 h. A signficiantly reduced viability and increase in the population of apoptotic dead cells were observed in GBM cells treated with the combination of Simva‐TMZ compared to cells treated with TMZ alone. Addition of MEV, FPP, GGPP and cholesterol did not attenuate these effects significantly. Sima‐TMZ treatment did not alter the total cholesterol pool in U87 and U251 cells compared to controls. Western blot analysis, immunocytochemistry and transmission electron microscopy revealed that the combination of Simva with TMZ inhibited autophagic flux. Overall, the results suggest that sensitization of GBM cells to TMZ‐induced apoptosis by Simva is independent on the cholesterol biosynthetic pathway but may involve inhibition of autophagy flux. Support or Funding Information Supported by funding from a Mitacs Accelerate PDF Award &amp; a Health Science Center Foundation Operating Grant. This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal .

Please cite this article as follows: Mirzaei N, Sepehri Z, Ghavami S. Autophagy, a possible future approach for tuberculosis treatment Int J Basic Sci Med. 2016;1(1):1-3. doi:10.15171/ijbsm.2016.01.

Wang, et al. have provided new and novel evidence about the importance of using α-fetoprotein (ALP) and glypican-3 (GPC-3) mRNA detection in peripheral blood of the patients with hepatocellular carcinoma (HCC) recurrence after ortothopic liver transplantation (OLT) [1]. HCC keeps on as major universal health apprehension ensuing in more than 1 million caner associated deaths annually [2][3]. These results are frankly attributable to a commonly pitiable prognosis for patients with HCC, with few attaining 5-year survival [3]. Although the incidence of HCC in North America has been comparatively low, a stable augmentation has taken place since the 1980s, while the prevalence of most other cancers has turned down [4]. Because of the comparative resistance of HCC to systemic chemotherapy, pioneering treatments such as transarterial chemoembolization and yttrium 90-labeled radiotherapy has been extended to improve disease manage. HCC often happens in the setting of cirrhosis, total hepatectomy with OLT affords one of the only prospective cures for both diseases so at present liver transplantation has been established as a cure of patients with HCC and associated cirrhosis. In spite of this, the return of HCC after liver transplantation remains an unanswered concern. Mazzaferro et al. proved that a stringent assortment of HCC patients, focusing on tumor-staging before transplantation, could facilitate an acceptable tumor-free survival. They also found poor outcome among patients whose tumors did not descend within the criteria (i.e., the Milano criteria) [5]. In general, immunosuppressed organ allograft recipients are at increased jeopardy of developing a number of malignancies. Vivarelli et al. [6] confirmed that the amount of immunosuppressant agents (cyclosporine) administered was associated with HCC recurrence after liver transplantation. Recent studies showed that the risk of HCC recurrence subsequent liver transplantation is larger with antibody treatment than with other classes of immunosuppressive agents [7]. Several researches have been performed to find a reliable tumor marker for HCC and HCC recurrence after OLT. During the past 50 years, AFP has been used as a serum tumor marker for HCC, but its function as a probable predictive factor in OLT for HCC has not been established. AFP is a major transport protein in the fetus [8] and being secreted in only about 70% of HCC cases, so both false-negative and false-positive rates are elevated when considering AFP as the serological marker for the detection of HCC [9]. In general, patients have an initially high serum AFP level when HCC is diagnosed. Serum AFP level is also considered an effective tumor marker in the surveillance of HCC recurrence [10]. It is expected that recurrent HCC to have the identical bioactivity as the primary tumor and to exude high levels of AFP, making AFP an apparently reliable choice. Regular checks of serum AFP levels in patients who originally had high AFP levels to recognize HCC recurrence is suggested as a practice guideline by the National Comprehensive Cancer Network (NCCN) version 1, 2009. The whole AFP can be alienated into three different sub-fractions, AFP-L1, AFP-L2, and AFP-L3, based on its reactivity to LCA on affinity electrophoresis. AFP-L1 does not react with LCA. It is increased in chronic hepatitis and liver cirrhosis and comprises a bulk fraction of the total AFP in non-malignant liver diseases. AFP-L2 is mostly derived from yolk sac tumors and could also be detected in maternal serum during pregnancy. AFP-L3 is the LCA-bound fraction of AFP. It has been reported that malignant liver cells produce AFP-L3, still when HCC is at its near the beginning stages [11]. AFP-L3 has been established to be a marker for HCC with a high specificity of 95%. AFP-L3-secreting hepatocytes have an increased tendency for rapid growth, early attack, and intrahepatic metastasis, thus leading to a poorer prognosis in affected individuals [12][13]. AFP and AFP-L3 levels decline to normal levels with effective therapy, and then rising levels suggest disease progression or recurrence. Therefore combination of total AFP and AFP-L3 could be a very reliable biomarker for HCC and HCC recurrence in OLT. GPC-3 (also called DGSX, GTR2-2, MXR7, OCI-5, SDYS, SGB, SGBS, and SGBS1), a cell exterior protein, is extremely expressed in HCC and some other human cancers including melanoma [14]. The GPC3 gene encodes a 70-kDa-precursor core protein, which can be sliced by furin to make a 40-kDa amino (N) terminal protein and a 30-kDa membrane-bound carboxyl (C) terminal protein, which has two heparan sulphate (HS) glycan chains [15]. The GPC3 protein is attached to the cell membrane by a glycosyl-phosphatidylinositol (GPI) anchor [16]. GPC-3 is not expressed in hepatocytes of healthy subjects and patients with non-malignant hepatopathy, and can be detected in about 50% of HCC patients and 33% of HCC patients seronegative for AFP. The specificity of GPC-3 could be considered 100% according to some previous research [17]. Some clinical studies have indicated that the simultaneous determination of GPC-3 and AFP could significantly increase the sensitivity in HCC detection, without a reduction in the specificity [18]. The clinical significance of changes in tumor markers after therapy has been investigated for various solid tumors, and these changes are closely correlated with tumor response. For HCC, early studies have shown that AFP levels decrease rapidly after complete surgical resection and increase with tumor recurrence, suggesting that AFP response could potentially predict tumor response [19]. A few recent studies highlighted the potential role of serial AFP screening in the evaluation of treatment response, with the supposition that AFP is indicative of tumor growth and activity [20][21]. Regarding the new research combination of total AFP, AFP-L3, and GPC-3 could be a good marker for HCC recurrence after OLT. Therefore Wang et al paper in this volume pointed out a very good beginning to monitor HCC patients who have under-gone the OLT and needs monitoring for tumor recurrence.

Background: Induced pluripotent stem cells (iPSCs) have the ability to proliferate indefinitely and differentiate into three germ layers of ectoderm, mesoderm, and endoderm. Definitive induction is the first and the most delicate stage of differentiation of various iPSC-derived organs. It has been found that the Wnt signaling pathway implicates in embryogenesis, organogenesis, and cell communication. Objectives: In the present study, we aimed to investigate the expression pattern of the Wnt5a gene as an indicator of non-canonical Wnt signaling activity during definitive endoderm induction of iPSCs. Methods: Human iPSCs (RSCB0042) were acquired from Royan stem cell bank of Royan Institute (Tehran, Iran). The iPSCs were cultured on a feeder layer of mitomycin-inactivated mouse embryonic fibroblasts (MEF), and iPSC colonies were collected for embryoid body (EB) generation by suspension culture method. Then endoderm induction step was performed using a series of small molecules. The quantitative real-time PCR was used to assess the mRNA expression of wnt5a, Nanog, OCT4, SOX17, and FOXA2 genes. Results: The production of efficient EBs confirmed by a decrease in Nanog and Oct4 gene expression and the success of DE (definite endoderm) induction step was confirmed by a high expression level of DE specific genes, Sox17, and FoxA2. A significant upregulation of Wnt5a in EB samples and a minor decrease at day 4 was observed. However, the differentiation process followed by an incremental fashion in Wnt5a mRNA expression starting from day 4 of differentiation among the samples of days 6 and 8 (DE stage). Conclusions: Our results suggest that Wnt5a is more activated at the later steps of endoderm induction rather than the early steps, which may be due to the stimulation of canonical Wnt signaling. Finding the expression level of Wnt5a could rise insights for developing more efficient differentiation induction protocols.

Several studies have reported a possible association of miR-146a rs2910164 polymorphism with Breast Cancer (BC) development. However, the correlation between this polymorphism and susceptibility to BC is under debate. The current meta-analysis was designed and performed to more conclusively evaluate the miR-146a rs2910164 polymorphism and its potential link to BC.Our team has selected eligible studies (published up to October 2, 2020) from several electronic databases, including Web of Science, PubMed, Scopus and Google Scholar. A total number of 9,545 BC cases and 10,030 controls extracted from 26 eligible articles were included in this study. We utilized pooled Odds Ratios (ORs) as well as 95% confidence intervals (95% CIs) under five genetic models for quantitative estimation of any possible association between miR-146a rs2910164 polymorphism and BC.Based on this meta-analysis, our findings suggest that there is no significant association between miR-146a rs2910164 polymorphism and BC risk. However, stratified analysis revealed that the rs2910164 polymorphism significantly increased the risk of BC in hospital-based studies using the homozygous genetic model (OR=1.37, 95%CI=1.01-1.86, p=0.043, CC vs. GG). Neither Asian nor Caucasian populations showed any significant association between rs2910164 polymorphism and BC susceptibility.In summary, our findings suggest that BC development is not associated with miR-146a rs2910164 polymorphism. However, larger ingenious future investigations might be needed for a more precise estimation of any association between miR-146a rs2910164 polymorphism and BC.

Zn (II) is an important regulator of caspase-3, as well as an antioxidant, microtubule stabilizer, growth cofactor, and anti-inflammatory agent. Over the past 30 years, many researchers have demonstrated the important role of Zn (II) in a variety of physiological processes, including growth and development, maintenance and priming of the immune system, and in tissue repair and regeneration. In this study, we present evidence that chelation of extracellular zinc by diethylenetriaminepentacetic acid (DTPA) in different concentrations causes cell death in carcinoma cell lines, HT29/219 and SW742. Hoechst 33258 staining revealed that cell death was mainly by apoptosis. Additionally, significant increases in the activity of caspase-3 and -9 were observed in both cell lines. Caspase-8 activation was negligible in both cell lines. The cytotoxicity/apoptotic effect of DTPA was inhibited significantly by Zn (II), Cu (II) and N-Acetyl-L-Cysteine (NAC) (P<0.05). Therefore, DTPA, the membrane-impermeable metal ion chelator, induces apoptosis through the depletion of extracellular

Following myocardial infarction (MI), relatively quiestent cardiac fibroblasts undergo phenoconversion to active, hyper‐synthetic myofibroblasts. During the late stages of wound healing, myofibroblasts are removed from the infracted region through apoptosis thus decreasing scar cellularity. Apoptosis and autophagy regulate cell fate. The role of apoptosis within the remodeling heart has been well described however the role that autophagy plays following an MI is relatively unknown. Ski is a negative regulator of TGF‐β signaling and has been implicated to play a role in the post‐MI heart. Herein we investigated the regulatory role that Ski has on apoptosis and autophagy using first passage primary rat cardiac myofibroblasts. Using an adenoviral approach for gene delivery, both MTT and Live/Dead viability/cytoxicity assays demonstrated a reduction in the number of viable cells and an increase in the number of dying cells due to the over‐expression of Ski. Additionally, over‐expression of Ski led to distinct morphological changes characteristic of apoptosis. These findings were confirmed via Western blot analysis, and caspase GLO analysis showing induction of executioner caspases‐3 and ‐7, as well as the mitochondrial regulator Bax. Furthermore, evidence of caspase‐9 cleavage, but not caspase‐8, indicated that this was primarily an intrinsic apoptotic response. Markers for autophagosome formation including LC3‐B and ATG‐7 were found to be significantly reduced following Ski over‐expression. In summary, we found that over‐expression of Ski in primary cardiac myofibroblasts leads to an induction of the intrinsic apoptotic pathway and reduction in autophagy leading to cell death. Grant Funding Source : Canadian Institutes of Health Research

Cardiac fibrosis is linked to fibroblast-to-myofibroblast phenoconversion and proliferation but the mechanisms underlying this are poorly understood. Ski is a negative regulator of TGF-β-Smad signaling in myofibroblasts, and might redirect the myofibroblast phenotype back to fibroblasts. Meox2 could alter TGF-β-mediated cellular processes and is repressed by Zeb2. Here, we investigated whether Ski diminishes the myofibroblast phenotype by de-repressing Meox2 expression and function through repression of Zeb2 expression. We show that expression of Meox1 and Meox2 mRNA and Meox2 protein is reduced during phenoconversion of fibroblasts to myofibroblasts. Overexpression of Meox2 shifts the myofibroblasts into fibroblasts, whereas the Meox2 DNA-binding mutant has no effect on myofibroblast phenotype. Overexpression of Ski partially restores Meox2 mRNA expression levels to those in cardiac fibroblasts. Expression of Zeb2 increased during phenoconversion and Ski overexpression reduces Zeb2 expression in first-passage myofibroblasts. Furthermore, expression of Meox2 is decreased in scar following myocardial infarction, whereas Zeb2 protein expression increases in the infarct scar. Thus Ski modulates the cardiac myofibroblast phenotype and function through suppression of Zeb2 by upregulating the expression of Meox2. This cascade might regulate cardiac myofibroblast phenotype and presents therapeutic options for treatment of cardiac fibrosis.

Cardiovascular diseases (CVD) leading to heart failure are associated with varying degrees of myocardial cell loss and cardiac fibrosis. Cardiac fibrosis is prevalent, being found in conditions such as myocardial infarction, ventricular hypertrophy due to hypertension or valvular heart disease, or diabetic cardiomyopathy. Furthermore, there are limited therapeutic options for its treatment. HMG-CoA Reductase inhibitors ("statins") have been shown to limit cardiovascular events in patients with CVD through modulation of lipid profile. HMG-CoA Reductase inhibition downregulates the production of the cholesterol precursor L-mevalonate. Mevalonate depletion has been shown to induce autophagy, apoptosis, and endoplasmic reticulum stress in various cell types. However it is unclear if this is a class effect or a phenomenon specific to certain compounds. We hypothesize that lipophilic statins induce cell death in primary hATF via mevalonate depletion; whereas hydrophilic statins do not have same effect on hATF viability. The protocol received IRB approval, and hATF were isolated from the atrial appendage collected from consented patients undergoing open heart surgery and were cultured in standard conditions. Cells were treated with atorvastatin, simvastatin (lipophilic statins) or pravastatin (hyodrophilic statin) 0.1, 1.0 or 10 µM for 24, 48, 72 or 96 hours. Cell viability was assessed using an optical MTT assay. Expression of key apoptotic and autophagy protein markers was assessed by western blot technique. Rescue of viability was achieved through coincubation with mevalonate. hATF treated with 0.1 - 10 µM atorvastatin or simvastatin (lipophilic statins) showed progressively reduced cell viability in time and dose-dependent manner; which was associated with expression of key apoptotic cascade proteins Bcl2, Bax and cleaved caspase 3, showing a clear induction of apoptosis. Treated hATF also showed an increase in Atg5-12 expression at 24 hour, indicating an induction of early autophagic response. Viability could be rescued by coincubation with mevalonate or pretreatment with Ikk-β adenoviral vector. Treatment with pravastatin (hydrophilic statin) did not affect cell viability or expression of autophagy and apoptosis markers. We conclude that statins induce cell death in hATF which is mediated by mevalonate depletion through HMG-CoA reduactase inhibition. Lipohilic statins impair the viability of primary hATF in culture by induction of intrinsic apoptotic pathways whereas hydrophilic statins do not affect hATF viability. This could be due to the hydrophilic characteristic of the molecule, preventing its uptake by the cell. This may represent an additional pleiotropic effect of stains independent of its effect on lipid metabolism.

Background: Most severe asthmatics exhibit steroid refractory disease that limits effectiveness of therapy and typically associ ated with chronic neutrophilia, and release of a unique repertoire of “steroid refractory” mediators. Thus, a better understanding of severe disease, including triggers of these steroid refractory mediators is critical to address unmet clinical needs. Our aim was to identify whether S100A9 knock-out mice (the subunit that stabilizes the functional S100 protein heterodimer) elicits attenuated hallmarks of asthma including, airway inflammation (neutrophilia and eosinophilia), airway hyperresponsiveness and airway remodelling in a murine model of allergic asthma. Methods: Eight week old Balb/c mice were subjected to intranasal (i.n) challenges with house dust mite (HDM) (Dermatophagoides pteronnyssinus) extract (0.7mg/mL in saline) - acute (5 consecutive daily i.n challenge for 2weeks) and chronic (3 daily mid-week HDM i.n challenge for additional 5 weeks). Animals were assessed for S100A8/A9 at: ~8hrs and 48hrs post acute protocol (peak neutrophilia and eosinophilia respectively); and 48 hrs post chronic protocol (established airway remodeling). We also assessed baseline respiratory mechanics in S100A9 knockout (KO) mice via flexiVent. Using Hu man endobronchial biopsies (severe asthma) and patient-matched bronchoalveolar lavage fluid (BALF), we also performed immunohistochemistry and Enzyme Linked ImmunoabSorbent Assay (ELISA) to assay S100A8/A9 and Receptor for Advanced Glycation Endproducts (RAGE) expression. Results: Expression of S100A8/A9 and its primary receptor, RAGE, are increased in asthmatic biopsies compared to healthy biopsies. S100A8/A9 release in BALF fluid was elevated by 6 fold in human asthmatic [605 ng/ml (24-2420)] compared to healthy subject BALF [107 ng/ml (38-400)]. In mice, acute HDM challenge induces and sustains S100A8/A9 release in BALF, and this tracks with neutrophil cell numbers in BALF. S100A8/A9 and RAGE from lung tissue is increased in acute and chronic HDM challenged mice compared to naive controls. Using the acute HDM protocol, HDM S100A9 KO mice elicit reduced airway resistance (AR), tissue resistance (TR) and tissue elastance (TE) to methacholine challenge (6-50 mg/ml) compared to HDM wild-type mice. Similarly basal AR and TE are both reduced in the HDM S100A9 KO mice compared to HDM wild-type mice. Furthermore, total BALF cells including eosinophils and neutrophils appear reduced in S100A9 KO mice for both naive and HDM groups. Conclusion: In severe asthma and murine models of allergic asthma there is an elevated release and expression of S100A9/ A9 and RAGE. Improved lung mechanics and attenuated lung inflammation in S100A9 KO mice suggest that sustained levels of S100A8/A9 may contribute to disease pathogenesis.

Cardiovascular diseases (CVD) leading to heart failure are associated with myocardial cell loss and cardiac fibrosis. Hydroxymethylglutaryl-Coenzyme-A Reductase (HMGR) inhibitors ("statins") are widely used to limit cardiovascular events in patients with hypercholesterolemia and CVD by altering their lipid profile. HMGR inhibition reduces cholesterol precursor L-mevalonate production, whose depletion induces autophagy, apoptosis, and endoplasmic reticulum stress in various cell types. However it is unclear if this is a class effect or a phenomenon specific to various compounds. We examined the in vitro effect of HMGR inhibition on human atrial fibroblast (hATF) viability with particular reference to hydrophilic vs lipophilic compounds. Hypothesis- Lipophilic statins induce cell death in primary hATF via mevalonate depletion; whereas hydrophilic statins do not have any effect on hATF viability. IRB approval was obtained for collection of hATF from consenting patients undergoing open heart surgery. Cells were treated with atorvastatin, simvastatin or pravastatin (0.1, 1.0 or 10 λM) for 24, 48, 72 or 96 hours. Expression of proteins involved in the regulation of apoptosis and autophagy was assessed using immunoblotting. Cell viability was assessed using MTT assay. Treatment of hATF with 0.1 - 10 λM atorvastatin or simvastatin (lipophilic statins) resulted in progressively reduced cell viability in time and dose-dependent manner. Viability could be rescued by coincubation with mevalonate. Expression of key apoptotic cascade proteins -Bcl2, Bax and cleaved Caspase3 showed a clear induction of apoptosis. Also, there was an increase in Atg5-12 expression at 24h indicating induction of early autophagic response. Pravastatin (hydrophilic statin) did not affect cell viability or autophagy and apoptosis. We conclude that statin-induced cell death is mediated by mevalonate depletion, which activates intrinsic apoptotic pathways in hATF. Lipophilic statins impair the viability of hATFs in vitro, whereas hydrophilic statins have no effect on cell growth and cell viability of hATFs. This may represent an additional pleiotropic effect of statins, and may represent a novel therapeutic option for the prevention and treatment of cardiac fibrosis.

Cardiac fibroblasts phenoconvert to hypersecretory myofibroblasts and deposit matrix resulting in cardiac fibrosis and autophagy that is associated to fibroproliferative events. Autophagy inhibition was studied in both primary (P0) rat cardiac fibroblasts (PF) and mouse embryonic fibroblasts (MEF). Autophagy was inhibited in PF using Baf‐A1 or 3‐MA while MEF ATG3 and ATG5 KO were also used. Western blot analysis, Immunofluorescence, and transmission electron microscopy were used to investigate autophagy and fibroblast to myofibroblast phenoconversion. α‐SMA and ED‐A fibronectin were assessed as phenoconversion markers. Inhibition of autophagy was revealed by punctate LC3‐II staining and double membrane autophagosomes in combination with decreased myofibroblast markers in PF vs. controls. Inhibition of autolysosome formation and accumulation of autophagosomes were found in Baf‐A1 treated cells. Conversely, ATG3 and ATG5 KO MEF cells showed increased myofibroblast markers vs. WT controls. Increased α‐SMA was seen in MEF ATG3 and ATG5 KO vs. controls while inhibition of autophagy in PFs showed diminished myofibroblastic phenotype vs. controls. A link between fibroblast phenoconversion and autophagy is established. This effect is dependent on cell type eg, PFs vs. MEFs. This finding illustrates that autophagy functions in a variable manner in differentiated vs. non‐differentiated fibroblasts.

Dietary trans fatty acids (TFA) are linked to cardiovascular disease which has led to recommendations to remove TFAs from diet. We investigated the effects of two common fatty acids ‐ vaccenic acid (VA ‐ found in dairy products), and elaidic acid (EA ‐ found in hydrogenated vegetable oils) on cell death in primary rat ventricular myofibroblasts (rVF). VA‐ and EA‐treated cells exhibited apoptotic cell death in rVF, as reflected in MTT and FACS assays. A significant reduction in mitochondrial membrane potential and an increase in the Bax/Bcl2 ratio and Bax translocation to the mitochondria was found suggesting apoptosis in the presence of TFA. Elevated cleaved caspase‐9 and caspase‐3 (without caspase‐8), shows that EA and VA treatments induce caspase‐dependent apoptosis. Transmission electron microscopy reveals autophagosome formation and lysosome activation in VA and EA treated rVF. Induction of autophagy markers including LC3‐B lipidation, increased Atg5‐12 formation and increased beclin‐1 were observed confirming autophagy activation by EA and VA. Inhibition of ATG3 and ATG5 significantly inhibited cytotoxic effects by preventing caspase‐3 and caspase‐7 activation in EA and VA treated rVF. In conclusion we show that TFAs induce apoptosis and autophagy in rVF and that TFA‐induced autophagy is required for this proapoptotic effect. Supported by CIHR and DFC.

Tumor necrosis factor alpha (TNFα) is one of the main mediators of inflammatory response in many pathological diseases, involved in a widespread biological functions, including autophagy. Previous data obtained in our laboratory demonstrated that TNFα and some autophagy markers (which markers please indicate) are overexpressed in a severe inflammatory disease such as vernal keratoconjunctivitis (VKC). In the present study we explored the role of TNFα in the induction of autophagy in VKC, using an in vitro model. Primary conjunctival cell cultures were treated with TNFa and analysed by qPCR and western blotting for expression of some autophagy and lysosomial markers at 4, 10 and 24 hours after exposure. qPCR results demonstrated that LC3B, Beclin-1, LAMP1 and p62 strongly increased from 4 to 24 hours, whereas the expression of Catepsin D, a protein implicated in lysosomial apototic pathway, was comparable to that of untreated control. Western blotting analysis revealed lipidation of LC3B quantified as an increased LC3BII/LC3BI ratio. Moreover, double immunofluorescence for Cathepsin D and LAMP1 showed that Cathepsin D was localized within the lysosomes at 4, 10, 24 hours after cell exposure to inflammatory stimuli. In conclusion, our data demonstrated that TNFα significantly induce in VKC LC3B lipidation, LC3BII/LC3BI ratio and p62 (qPCR) in the cells exposed to inflammatory stimuli which shows possible activation of autophagy pathway.

We investigated the structure and properties of PCL 10 nanofiber, PCL 5 /PCL 10 core-shell type nanofibers, as well as PCL 5 /PCL Ag nanofibres prepared by electrospinning. For the production of the fibre variants, a 5-10% solution of polycaprolactone (M w = 70000-90000), dissolved in a mixture of formic acid and acetic acid at a ratio of 70:30 m/m was used. In order to obtain fibres containing PCL Ag 1% of silver nanoparticles was added. The electrospin was conducted using the above-described solutions at the electrostatic field. The subsequent bio-analysis shows that synthesis of core-shell nanofibers PCL 5 /PCL 10 , and the silver-doped variant nanofiber core shell PCL 5 /PCL Ag by using organic acids as solvents is a robust technique. Such way obtained nanofibres may then be used in regenerative medicine for extracellular scaffolds: ( i ) for controlled bone regeneration due to the long decay time of the PCL, ( ii ) and as carriers of drug delivery nanocapsules. Furthermore, the used solvents are significantly less toxic than the solvents for polycaprolactone currently commonly used in electrospin, like for example chloroform (CHCl 3 ), methanol (CH 3 OH), dimethylformamide (C 3 H 7 NO) or tetrahyfrofurna (C 4 H 8 O), hence the presented here electrospin technique may allow for the production of multilayer nanofibres more suitable for the use in medical field.

We investigated the structure and properties of PCL 10 nanofiber, PCL 5 /PCL 10 core-shell type nanofibers, as well as PCL 5 /PCL Ag nanofibres prepared by electrospinning. For the production of the fibre variants, a 5-10% solution of polycaprolactone (M w = 70000-90000), dissolved in a mixture of formic acid and acetic acid at a ratio of 70:30 m/m was used. In order to obtain fibres containing PCL Ag 1% of silver nanoparticles was added. The electrospin was conducted using the above-described solutions at the electrostatic field. The subsequent bio-analysis shows that synthesis of core-shell nanofibers PCL 5 /PCL 10 , and the silver-doped variant nanofiber core shell PCL 5 /PCL Ag by using organic acids as solvents is a robust technique. Such way obtained nanofibres may then be used in regenerative medicine for extracellular scaffolds: ( i ) for controlled bone regeneration due to the long decay time of the PCL, ( ii ) and as carriers of drug delivery nanocapsules. Furthermore, the used solvents are significantly less toxic than the solvents for polycaprolactone currently commonly used in electrospin, like for example chloroform (CHCl 3 ), methanol (CH 3 OH), dimethylformamide (C 3 H 7 NO) or tetrahyfrofurna (C 4 H 8 O), hence the presented here electrospin technique may allow for the production of multilayer nanofibres more suitable for the use in medical field.

Rhabdomyosarcoma (RMS) is a muscle‐derived tumor and is the most common pediatric soft tissue sarcoma representing 5% of all childhood cancers. RMS is a major clinical problem in pediatric oncology. Treatment of RMS with the oral alkylating agent temozolomide (TMZ), alone or in combination with other drugs, has recently received considerable interest. However, the mechanism of action of TMZ remains unclear. The aim of this investigation was to determine if autophagy modulates TMZ‐induced cell death in RMS cell lines. MTT assay and Nicoletti flow cytometry analysis were used to measure cell death and apoptosis in SJCRH30 (RH30) human RMS cells and the non‐transformed mouse myoblast cell line C2C12, following TMZ treatment (100 μM). We monitored autophagy using transmission electron microcopy (autophagosome and autophagolysosome formation) and immunoblotting (LC3 lipidation, Atg15‐12 conjugation, p62 degredation). The effect of bafilomycin A1 on TMZ‐induced cytotoxicity was also evaluated. We showed that TMZ decreased the viability of RMS cells in a dose/time‐dependent manner and induced accumulation of sub‐G 1 cell population, representing apoptotic cells. Interestingly, TMZ induced apoptosis by 17‐fold in the RH30 cells, but only increased apoptosis by 2.7‐fold in C2C12 cells. In RH30 cells, TMZ decreased the expression of antiapoptotic proteins Bcl‐XL and Mcl‐1, and increased the expression of the death gene Nix. Moreover, we showed that TMZ altered biochemical markers of autophagy, such as LC3 lipidation, Atg5–12 conjugation, and p62 degradation, and induced morphological evidence autophagy, including accumulation of autophagosomes and autophagolysosome, determined by transmission electron. Finally, treatment of RMS cells with the autophagy flux inhibitor (Bafilomycin A1, 4 and 6 nM) had no significant effect TMZ‐induced cell death in either cell line. In conclusion, our investigation showed that TMZ induced simultaneous autophagy and apoptosis in both RH30 and C2C12 cells; however, cell death induction by TMZ does not appear to be dependent on Bafilomycin A1 inhibited processes, such as autophagosome‐lysosome fusion or autolysosome acidification.

Autophagy is an intracellular self-degradative homeostasis process which eliminates undesirable and harmful macromolecules and organelles. Autophagy is also involved in self-renewal and differentiation of induced pluripotent stem cell (iPSCs).In this study, we investigated the expression profile of autophagy marker genes in human iPSCs during their differentiation induction toward insulin producing β-like cells.Human iPSC line, R1-hiPSC1, was used for differentiation induction toward β-like cells. The mRNA expression of Nanog, OCT4 (pluripotency markers), SOX17, FOXA2 (endodermic markers), PTF1A, NKX6.1 (exocrine/endocrine determinants), and PDX1 were measured during differentiation stages. Autophagy was monitored by genes expression study of four autophagy markers, MAP1LC3B, BECN1, SQSTM1/P62 and ATG5, along with protein expression profile of LC3b-II during differentiation stages.The mRNA expression measurement of pluripotency, endoderm and exocrine/endocrine marker genes confirmed that hiPSCs skipped pluripotency, differentiated into endoderm, passed through the pancreatic lineage commitment stage and successfully generated insulin producing β-like cells. Expression profile of autophagy genes during differentiation stages indicated the decreased expression levels at the early stages (EB and MEI) and then increased at the definitive endoderm stages (DEI 1, DEI 2 and DE) followed by a subtractive pattern toward the end of differentiation. The results of protein expression of LC3b-II were consistent with gene expression data.This study demonstrated the high contribution of key autophagy genes/proteins during the differentiation of hiPSC toward β-like cells. The enhanced autophagy levels were a prominent feature of early stages of differentiation and DE rather than the later stages.

Lipotoxicity is a form of cellular stress caused by the accumulation of lipids resulting in mitochondrial dysfunction and insulin resistance in muscle. Previously, we demonstrated that the mitophagy receptor BNIP3L/Nix is responsive to lipotoxicity and accumulates in response to a high-fat (HF) feeding. To provide a better understanding of this observation, we undertook gene expression array and shot-gun metabolomics studies in soleus muscle from rodents on an HF diet. Interestingly, we observed a modest reduction in several autophagy-related genes. Moreover, we observed alterations in the fatty acyl composition of cardiolipins and phosphatidic acids. Given the reported roles of these phospholipids and BNIP3L in mitochondrial dynamics, we investigated aberrant mitochondrial turnover as a mechanism of impaired myocyte insulin signaling. In a series of gain-of-function and loss-of-function experiments in rodent and human myotubes, we demonstrate that BNIP3L accumulation triggers mitochondrial depolarization, calcium-dependent activation of DNM1L/DRP1, and mitophagy. In addition, BNIP3L can inhibit insulin signaling through activation of MTOR-RPS6KB/p70S6 kinase inhibition of IRS1, which is contingent on phosphatidic acids and RHEB. Finally, we demonstrate that BNIP3L-induced mitophagy and impaired glucose uptake can be reversed by direct phosphorylation of BNIP3L by PRKA/PKA, leading to the translocation of BNIP3L from the mitochondria and sarcoplasmic reticulum to the cytosol. These findings provide insight into the role of BNIP3L, mitochondrial turnover, and impaired myocyte insulin signaling during an overfed state when overall autophagy-related gene expression is reduced. Furthermore, our data suggest a mechanism by which exercise or pharmacological activation of PRKA may overcome myocyte insulin resistance. <b>Abbreviations:</b> BCL2: B cell leukemia/lymphoma 2; BNIP3L/Nix: BCL2/adenovirus E1B interacting protein 3-like; DNM1L/DRP1: dynamin 1-like; FUNDC1: FUN14 domain containing 1; IRS1: insulin receptor substrate 1; MAP1LC3A/LC3: microtubule-associated protein 1 light chain 3 alpha; MFN1: mitofusin 1; MFN2: mitofusin 2; MTOR: mechanistic target of rapamycin kinase; OPA1: OPA1 mitochondrial dynamin like GTPase; PDE4i: phosphodiesterase 4 inhibitor; PLD1: phospholipase D1; PLD6: phospholipase D family member 6; PRKA/PKA: protein kinase, AMP-activated; PRKCD/PKCδ: protein kinase C, delta; PRKCQ/PKCθ: protein kinase C, theta; RHEB: Ras homolog enriched in brain; RPS6KB/p70S6K: ribosomal protein S6 kinase; SQSTM1/p62: sequestosome 1; YWHAB/14-3-3β: tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein beta

In article 1900113 Mohsen Akbari and co-workers develop a hydrogel-based mesh containing temozolomide-loaded poly(lactic-co-glycolic acid) microspheres with high encapsulation efficiency (GlioMesh) that can release anticancer drugs directly at a tumor site. GlioMesh can conform to the irregular structure of brain tissue because of its flexibility and release drugs for up to two months.

3604 Recently a novel defensin, Brevinin-2R, a 25-amino acid polypeptide was isolated from the skin of the frog Rana ridibunda. The novel molecule kills semi-selectively cancer cells, but does not exhibit the typical hemolytic activity for the majority of the Brevinin-family defensins. The anti-cancer activity was demonstrated on various malignant cells such as Jurkat (T-cell leukemia), BJAB (B-cell lymphoma), SW742 (colon carcinoma), and MCF-7 (breast adenocarcinoma). Also data have shown that L929 and MCF-7 cells stably transfected with a dominant-negative mutant of the pro-apoptotic Bcl-2-family member BNIP3 (DTM-BNIP3) were significantly more resistant towards Brevinin-2R triggered cell death. Furthermore, we observed decrease in mitochondrial membrane potential (ΔΨm) and that induction of cell death is independent of caspase activation. Over-expressing of Bcl-2 in cancer cell lines significantly counteracted ΔΨm triggered by Brevinin-2R. Malignant hematopoietic cell lines were more sensitive towards Brevinin-2R than primary peripheral blood mononuclear cells (PBMC). Our results demonstrate that the polypeptide leads to activation of the mitochondrial death pathway, but does not induce cell death in cancer cells by classical apoptosis. The semi-selective anti-cancer effect of Brevinin-2R makes it a promising lead molecule for the development of anti-cancer drugs.

[This corrects the article DOI: 10.18632/oncotarget.25324.].

Background: Autophagy is a vital cell survival mechanism that authorizes cells to assort to metabolic stress and is essential for the development and maintenance of cellular and tissue homeostasis, as well as the prevention of human disease. It has also been shown that autophagy plays a significant role in the development and differentiation of stem cells, as well as induced pluripotent stem cells (iPSCs). Objectives: The present study aimed to examine the mRNA expression of the ATG5 gene, one of the key markers of autophagy in human iPSCs (hiPSCs) during endoderm induction. Methods: In this study, we cultured the human iPSC line (R1-hiPSC1) on mitomycin-C, inactivated mouse embryonic fibroblasts (MEF) layer, and used hanging drop protocol to generate embryoid body (EB) and expose differentiation. The Real-time PCR method was used to examine the mRNA expression level of ATG5 in hiPSC during endoderm induction. Results: Our results demonstrated the high mRNA expression of ATG5 in the mesendoderm induction (MEI) stage, which shows the high rate of autophagy in MEI days rather than the other stages of differentiation. Conclusions: The modification of ATG5 gene expression within hiPSC during endoderm induction shows the importance of autophagy assessments in hiPSC differentiation. Therefore, subsequent studies are needed to clarify the details of autophagy effects on hiPSC differentiation.

Acute kidney injury (AKI) is an important complication of COVID-19 encompassing a wide range of presentations. SARS-CoV-2 is proposed to cause AKI in the patients through various mechanisms. We are, nevertheless, far from a comprehensive understanding of the underlying pathophysiological mechanisms of the kidney injury in this infection. AKI has been shown to be a marker of disease severity and also a negative prognostic factor for survival. Unfortunately, no effective preventive strategy to decrease the risk of kidney damage in these patients has yet been identified. In this hypothesis, we highlight the potential protective effects of acetazolamide, a carbonic anhydrase inhibitor, in preventing the proximal tubular damage caused by the virus through disrupting the virus-endosome fusion and also interfering with the lysosomal proteases. Our proposed mechanisms could pave the way for further in vitro studies and subsequent clinical trials.