第10卷，第4期

Asthma is a chronic lung disease that primarily affects the lower respiratory tract in more than 300 million people worldwide. Asthma is often triggered by exposure to certain substances or conditions, such as allergens or cold air. The overreaction to the allergens causes inflammation and narrowing of the airway, which leads to episodic or persistent symptoms including shortness of breath, nocturnal cough, chest tightness, and wheezing. The cause of the disease is believed to be a combination of genetic and environmental factors. Asthma can be managed with medications that reduce inflammation and keep the airway open, such as bronchodilators and corticosteroids. However, there is currently no ideal treatment for patients with severe asthma which involves airway remodeling (AR). AR is characterized by long-term modification of the airway structure, including airway smooth muscle hyperplasia, increased inflammatory cell number, and submucosal gland hypertrophy, leading to lung function decline. Thus, understanding the molecular mechanism underlying AR is critical for the development of novel asthma therapeutics.

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Herpesviruses are smart pathogens that can infect and have lifelong persistence inside the host. Many human herpesviruses have been shown to affect human health in immunologically challenged conditions. Mostly the herpesviruses involved in such cases and widely researched are herpes simplex virus (HSV), human cytomegalovirus (HCMV), Kaposi sarcoma herpesvirus (KSHV), and Epstein-Barr virus (EBV). All these herpesviruses share similar virion structures and life cycles. They possess dynamic mechanisms to manipulate host cell components for entry, immune evasion, replication, gene expression, maintenance of latency, or production of virions by the lytic cycle. Jean Beltran and colleagues reported that herpesviruses are unable to infect cells lacking peroxisomes. The peroxisome is a single membrane-bound organelle that is present in all eukaryotic cells. Their abundance, size, protein composition, and activity can vary according to cell type and stage. Organelle dysfunction related to biogenesis or organelle-driven cellular processes has been linked to developmental disorders, aging, neurological effects, and cancers.

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From an evolutionary perspective, the human cancer genome is based on an ancient gene module developed by the common ancestor of the Amoebozoa, Metazoa, and Fungi (AMF). The transition phase to multicellularity (premetazoan period) was a rather long period of evolutionary attempts and failures. During this time, suppressor genes were evolved to deactivate the G + S cell system inherited from the AMF ancestor, and anti-suppressor genes evolved to reactivate it in case of failed evolutionary attempts. These suppressor and anti-suppressor genes were the archetypes of tumor suppressors and oncogenes. Genes from the failed evolutionary attempts were not discarded but added as remnants to the so-evolved premetazoan genome. During the subsequent evolution of metazoans and animals, the reactivation of the premetazoan genome generates cancer. Animal cancers evolved many specific anti-host genes. The human cancer genome consists of (i) founder germ and soma G + S genes belonging to the AMF cell system, (ii) evolutionary failed gene packages from the premetazoan evolution, and (iii) specific anti-host genes evolved during animal evolution (specific anti-host genes). All these gene packages and regulatory gene networks generate the phenotypic heterogeneity of cancer and tumors.

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We read with great interest the manuscript "LPCAT1 functions as a novel prognostic molecular marker in hepatocellular carcinoma" by Zhang et al in a recent issue of Genes& Diseases. The authors conducted bioinformatics analyses using high throughput RNA sequencing data from TCGA to demonstrate that LPCAT1 is a novel and effective prognostic marker for hepatocellular carcinoma. We appreciate the contributions of the authors on the subject, nonetheless, we have some concerns that should be clarified in the following issues.

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Recently, Niculescu published an interesting paper concerning "deep homologies" between the formation of cancer stem cells by polyploidization and reproductive polyploidization of entamoeba. The life cycle of both systems shares the same genetic machinery inherited from the Last Eukaryote Common Ancestor (LECA). This theory claims that cancer is nothing more than the expression of a pre-existing life cycle that was reactivated in "sick and weakened metazoan cell that cannot continue its multicellular life and finds itself in the same dead-end situation". Proposed amoeba/cancer homologies actually comply with the theories claiming that cancer represents inverse recapitulation of metazoan differentiated cell evolution to the pro-metazoan stage. However, the real polyploidy formation, based on endoduplication, is not specific to cancer. There is, for example, polyploidization of normal hepatocytes and fibroblasts, especially in tissue regeneration. Endorduplication itself is a purely physiologic phenomenon during thrombopoiesis. In addition, this is not the only feature of the ancient single-cell eukaryote life cycle but is common to cancer and normal behavior of the primitive cell.

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We have carefully read the correspondence "False Discovery Rate Control in Cancer Biomarker Selection" authored by Li. The author clarified the need of paying attention to the control of false discovery rate (FDR) when screening for cancerbiomarkers. They expressed concerns about whether the 30 genes concluded in Figure 7 in our published paper "LPCAT1 functions as a novel prognostic molecular marker in hepatocellular carcinoma" call for FDR control. We would like to express our great gratitude to the author for his attention to the article published by our team. The questions raised by the author allowed us to deeply look into clinically relevant statistical issues. We would like to make the following explanations.

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ABT-199 (venetoclax) induces cell apoptosis in lymphoid malignancies mainly through the mitochondrial apoptosis pathway. However, long-term use of ABT-199 causes secondary drug resistance, which limits its use. Here, we show that the acquired resistance to ABT-199 in diffuse large B-cell lymphoma (DLBCL) cells is related to the upregulation of phosphorylated vascular endothelial growth factor receptor 2 (p-VEGFR2), BCL-XL and MCL-1. To identify determinants of resistance, we conducted ABT-199-resistant cell lines. We found that VEGFR2 drove the ABT-199 resistance in addition to the known involvement of the B-cell lymphoma 2 (BCL-2) family members. Notably, we also found that the ABT-199 resistance was associated with the increased production of reactive oxygen species (ROS) in ABT-199 resistant cells. Thus, combinatorial therapy with Apatinib and ABT-199 may offer a new approach to address ABT-199 resistance.

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Congenital heart disease (CHD) is the commonest birth defect, affecting approximately 9.4/1 000 live births. Atrial Septal Defect (ASD) is one of the commonest CHD clinical phenotypes, which frequently requires treatment either in childhood or adulthood, and can lead to severe complications such as right heart failure and cardiac arrhythmia. Previous genome-wide association studies (GWAS) have identified a region of chromosome 4p16 (Ch4p16) associated with the risk of ASD. The most strongly associated SNPs (rs870142, rs6824295 and rs16835979) lie within a 38.8-kb region of linkage disequilibrium encompassing only the long noncoding RNA STX18-AS1 (also named LOC100507266) .Associated SNPs are expression quantitative trait locus (eQTLs) for STX18-AS1 in adult ventricular myocardial tissue.

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EBV-positive diffuse large B-cell lymphoma (DLBCL), which has a clonal EBV carrying proliferation of B-cells, defines a new DLBCL subtype and predicts a poor prognosis. Further studies are needed to explore the underlying mechanisms in EBV lymphomagenesis. EBV encoded microRNAs (miRNAs) have been proven to contribute to the pathogenesis and oncogenesis of EBV-associated malignancies. In this study, research has focused on the pathological roles of miRNAs in EBV-positive DLBCL progression and survival, which might provide ideas for therapeutic decision to improve the prognosis.

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Here, we developed a comprehensive web-searchable database, designated as Human RNA Modifications Disease Database (HRMDD, http://bio-bigdata. hrbmu. edu.cn/HRMDD/home.jsp). RNA modification (RM) is an important mechanism of epigenetic regulation. With the evolution of both experimental technologies and computational methods, major progress has been made in identifying the genomic locations and distributions of various RM types throughout the transcriptome. Additional breakthroughs came from the identification and characterization of RM regulators. RMs are generally regulated by three different types of regulators, which are deposited, removed, and recognized by proteins known as "writers", "erasers" and "readers", respectively. Increasing evidence suggests that dysregulation of RMs and their regulators is implicated in various cancers, as well as other diseases. Despite these advances in understanding RMs and their regulators, their biological functions and mechanisms in human diseases remain largely unknown.

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While progenitor cell-based cardiomyocyte regeneration holds great promise of repairing an injured heart, primary cardiomyogenic progenitors (CPs) have a limited life span in culture, hampering the use of CPs for in vitro and in vivo studies. We previously isolated primary CPs from mouse E15.5 fetal heart, and reversibly immortalized them with SV40 large T antigen (SV40 LTA), resulting in immortalized CPs (iCPs), which maintain long-term proliferation and express cardiomyogenic markers and retain differentiation potential under appropriate differentiation conditions. The SV40 LTA plays essential roles in the simian virus 40 infection of permissive cells for the production of progeny virions, and in the infection of non-permissive cells, leading to malignant transformation or immortalization. While, as one of the most commonly-used immortalizing genes, SV40 LTA is thought to immortalize primary cells depending on its ability to complex with and hence inhibit p53 tumor suppressor, the exact mechanism underlying SV40 LTA-mediated immortalization remains unknown. It was shown that induced pluripotent stem cells (iPSCs) could be established from somatic fibroblasts by simultaneous introduction of the four transcription factors OCT4, KLF4, SOX2, and C-MYC (also known as iPS factors or Yamanaka factors), and that reduced p53 expression facilitated reprogramming process. We hypothesized that SV40LTA-mediated immortalization may at least partially resemble the reprogramming phenomenon. Using the most potent osteogenic factor BMP9, we demonstrated that the immortalized cardiomyocytes exhibited mesenchymal stem cell (MSC) characteristics and osteogenic differentiation potential.

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Nuclear encoded genes can cause early-onset mitochondria-related disorders such as Leigh or Leigh-like syndrome. Defects in DNAJC30 have been implicated in mitochondriarelated diseases such as Leber's hereditary optic neuropathy (LHON) and Williams syndrome (WS). However, the role of DNAJC30 in disease progression concerning mitochondrial dysfunction has yet to be fully understood. Here we report a 12-year-old boy with acute dystonia onset at age 10.Brain magnetic resonance imaging (MRI) showed bilateral basal ganglion and thalamic hyperintensities concerning putaminal necrosis. He then developed bilateral optic atrophy and rapid progressive bilateral visual loss. Exome sequencing analysis of his peripheral blood sample revealed a homozygous nonsense germline variant, c.24G> A (p. W8X) in the DNAJC30 gene, proven to result in a complete loss of DNAJC30 protein expression in cells. Remarkably, the same germline variant was then identified by whole genome sequencing in another unrelated patient, a 17-year-old male. He also showed acute bilateral optic atrophy, subacute central vision loss, and abnormal brain MRI.In vitro functional analysis further confirmed that DNAJC30 deletion inhibited cell growth and induced mitochondrial disorders, evidenced by decreased oxygen consumption rate (OCR), reduced mitochondrial membrane potential (ΔΨm), and increased cellular and mitochondrial reactive oxygen species (ROS), presumably due to decreased complex I enzyme activity. Collectively, a homozygous nonsense germline variant in DNAJC30 (c.24G> A) was identified to cause mitochondria-related disorders. This rare DNAJC30 pathogenic variant will be useful in their diagnosis/prognosis and highlights the significance of the roles of DNAJC30 in the maintenance of normal mitochondrial function and brain development.

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The genus Streptococcus, a gram-positive bacterium, contains species associated with immune activation at morbid outcomes, including scarlet fever, endocarditis, meningitis, and rheumatic fever. In both stool and saliva, there is a positive correlation between Streptococcus abundance and Sjogren's syndrome (SS) and systemic lupus erythematosus (SLE). Interestingly, the HLA class II DQB1*06: 02 allele is both associated with the risk of SLE and plays putative roles in Streptococcus biology such as mimicry of bacterial antigens and related host antigens, an event that is associated with a loss of tolerance. We hypothesized that the Streptococcus-SLE association might be mediated by thep resence of SLE-risk allele DQB1*06: 02. Similarly, intercellular adhesion molecule-1 (ICAM-1), also known as CD54, and B-cell activating factor (BAFF), also known as tumor necrosis factor ligand superfamily member 13B (TNFSF13B), are elevated in the serum of SLE patients. The serum protein levels of BAFF and ICAM1 may therefore further complicate the interplay among Streptococcus sp., DQB1*06: 02, and SLE.

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Adult neurogenesis occurs in two specialized regions of the mammalian brain, the subventricular zone (SVZ) and the subgranular zone (SGZ) of the dentate gyrus (DG). Adult hippocampal neural stem cells (NSCs), referred to as Type 1 cells represented by radial glia-like cells (RGLs), generate Type 2 cells that are divided into Type 2a and Type 2 b subpopulations, the latter of which give rise to Type 3 cells (neuroblasts). Type 3 cells exit the cell cycle and give rise to immature neurons that subsequently mature into functional granule cells. The neural regenerative capacity depends on maintaining a reservoir of NSCs and the integrity of the neurogenic niche, which are regulated by neurotransmitters and numerous signals including Notch, Wnt, and BMPs. Here we showed that BMP9 (a.k.a., GDF2) deficiency altered the dynamics of adult hippocampal neurogenesis as proliferative activity was decreased in Type 2a cells in Bmp 9 knockout (Bmp9KO) mice. Surprisingly, the number of cells committed to the neuronal lineage was comparable to wildtype while the label retention was increased in immature neurons, suggesting mitigation of reduced proliferation in Bmp9KO mice by increased cell survival. We also found Bmp9 was not expressed in the adult brain, precluding cell-autonomous regulation of neurogenic populations like other BMPs. Our findings thus demonstrate BMP9 regulates adult homeostatic neurogenesis.

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Gastric cancer (GC) is worldwide the sixth most diagnosed and third leading cause of cancer deaths, with poor and late prognosis, probably due to post-surgery adjuvant treatment resistance and lack of athorough panel of prognostic markers. We have previously shown that mitotic arrest deficient 2 (MAD2, encoded by MAD2L1), a key protein of the spindle assembly checkpoint, is relevant in GC cells; its interference impairs migration and growth, while its overexpression correlates with tumorigenesis. Here we show a similar correlation with overall survival (OS) in a pilot patient series. We hypothesized that MAD2 overexpression might relate to micro-RNA (miRNA) deregulation. Bioinformatic analysis identified miRNAs specifically targeting MAD2L1-3′UTR. Expression of miR-19a and miR-203 inversely correlated with MAD2L1 expression in GC cell lines and patients' samples. A broader analysis using Cancer Genome Atlas data showed that only high miR-19 levels correlated with a better OS, especially in patients overexpressing MAD2L1.In GC cells, miR-19a expression reduced cell migration and invasion capability and increased apoptosis, in combination with classical and new antitumoral drugs. We propose that miR-19a is a critical regulatorofMAD2 protein inGC, with potential clinical use as a prognostic biomarker, and as a model for MAD2 interfering agent design with therapeutical potential.

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Atherosclerosis is a chronic progressive disease and one of the major causes of cardio-cerebral vascular diseases. Accumulating evidence indicates that endothelial dysfunction is the initiating step in atherosclerosis. The pattern of local blood flow becomes disturbed (low and oscillatory shear stress[OSS]) in the curved or branched segments of the arterial tree, causing endothelial cells (ECs) to exhibit athero-susceptible phenotypes, such as hyperproliferation and inflammation. By contrast, laminar blood flow in the straight parts of the arterial tree (high and unidirectional laminar shear stress[LSS]) causes ECs to act in a way that protects against atherosclerosis. The short isoform of Tet methylcytosine dioxygenase 1 (TET1s) is implicated in early embryonic development, cancer, and the mammalian nervous system. However, whether TET1s expression in ECs can regulate the initiation and development of atherosclerosis has not been explored. In the present study, we provided in vivo and in vitro evidence that atherogenic OSS reduced TET1s expression in ECs. The reduction in TET1s expression in turn induced the inflammatory and proliferative responses of ECs and ultimately led to atherosclerotic lesion formation. In addition, we demonstrated that TET1s prevented the OSS-mediated nuclear translocation of yes-associated protein (YAP) by increasing its phosphorylation at serine 127.These findings support the protective role of TET1s in modulating endothelial athero-susceptible phenotypes and atherogenesis.

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Familial adenomatous polyposis (FAP) and MUTYH-associated polyposis (MAP) are colon cancer predisposition syndromes. FAP is an autosomal dominant inherited condition caused by germline mutations in the adenomatous polyposis coli (APC) gene and characterized by hundreds to thousands of colorectal adenomas. FAP can be classified into several clinical forms, including profuse FAP (> 1,000 adenomas), intermediate FAP (100-1,000 adenomas), attenuated FAP (AFAP) (< 100 adenomas), and gastric polyposis and desmoid FAP (GD-FAP) (< 50 adenomas).FAP patients also have an increased risk of extra-colonic manifestations. MAP is inherited in an autosomal recessive manner. Monoallelic MUTYH mutation carriers are at increased risk for colonic and extra-colonic cancer. Biallelic MUTYH mutations are associated with colorectal polyposis and an increased life-time risk of gastrointestinal cancers.

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Hepatocellular carcinoma (HCC), as one of the most common types of primary liver cancer (PLV), accounts for approximately 75%-90% of all PLV casesidentified worldwide. Currently, the most effective treatment for HCC patients constitutes liver transplantation. However, due to high rates of recurrence a poor prognosis is predicted. Therefore, accurate HCC biomarkers are urgently needed to develop innovative therapeutics. Most of earlier investigations to identify biomarkers have been severely limited by sample size, as those studies used only data sets generated on the same chip platform. Considering the decisive role of sample size, we aimed to identify and validate diagnosis and prognosis biomarkers associated with HCC based on the expression data of seven transcriptome microarray gene expression (MAGE) datasets from two different platforms, namely Illumina and Affymetrix. To discover differentially expressed genes (DEGs), a metaanalysis based on an empirical Bayesian approach (ComBat) was first applied to the gene expression profiles of 939 tissue samples. Irigoyen et al technique was used to preprocess and integrate the data. Data import, data filtering, and normalization for each experiment were carried out separately (Text S1.1). Then, using the bioin-formatics technique of weighted correlation network analysis (WGCNA), the highly associated prioritized DEGs (Text S1.2, S1.3) were clustered. To identify sub-clusters and confirm candidate genes, co-expression networks and topological analysis were applied (Text S1.4).Finally, for these genes, the least absolute shrinkage and selection operator (LASSO) regression model was used and validated to provide a diagnostic model (Text S1.5).To construct a prognostic model, the Cox proportional hazard regression analysis was applied and validated (Text S1.6).All statistical tests were two-sided (Text S1.7) and the results with an adjusted p-value≤0.05 were considered statistically significant (Text S1.8).The flowchart diagram is presented in Figure 1A. The comparative boxplots and PCA plots in Fig. S1AeD, confirm the efficacy of the ComBat for batch effect removal. From the meta-analysis, 292 genes were identified as DEGs, satisfying the criteria of the absolute value of log2-fold change (logFC) > 1 and false discovery rate (FDR) < 0.05. Moreover, to assess bias and reproducibility across microarray experiments, we modified a comparison of individual analyses using data from two platforms and a meta-analysis. The result of integrative meta-analysis after prioritization showed 239 common DEGs (Text S2.1 and Table S5).We found possible relationships between the expression profiles of 239 mutual DEGs using Pearson's correlation coefficients (PCCs).The hierarchical cluster tree and topological overlapping matrix were used to screen out cluster modules. DEGs were divided into four parts, of which blue, red, and turquoise modules were considered the most significant parts. Through the application of the molecular complex detection (MCODE) plug-in, the sub-clusters of differential co-expression module (DCEM) were found and visualized.

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The flow-through hybridization and gene chip (FHGC) was developed to used in clinical molecular diagnostics for thalassemia, human papillomavirus (HPV), and other diseases. FHGC could improve hybridization efficiency and reduce hands-on time, thus improving precision, reproducibility, and traceability. Multiple genotypes can be detected simultaneously without incurring high costs. During the experiment, the polymerase chain reaction (PCR) product or buffer forced through the membrane matrix, PCR products are apprehended by a probe attached to the membrane, and molecular typing of the disease or microorganism was determined after subsequent amplification by the streptomycin avidin (the details are given in the Supplementary File). However, the steps prior to hybridization, such as DNA extraction, PCR amplification, and sample loading into the hybridization wells, require a significant amount of manual operation. Each batch involves handling dozens of samples, and Eppendorf (EP) tubes used to load the amplification mixture are typically packed tightly together. Furthermore, the reagent is colorless and transparent, making it difficult to determine whether the sample was loaded into EP tube or not with a very small sample size. Operator carelessness or distraction will result in errors and unnecessary trouble.

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Recently, p. Glu1121Ter in PLXNA1 was identified as potential cause for a patient with parkinsonism. However, no further replication has been conducted in a wider range of Parkinson's disease (PD) cohorts. To evaluate the genetic association of PLXNA1 with PD, we systematically analyzed the rare protein-coding variants in 1,245 Chinese patients with whole exome sequencing. Fisher's exact test was performed between each variant and the risk of PD, while over-representation of rare variants in patients was examined with optimized sequence kernel association test at gene level. Totally, 42 rare variants were identified. At variant level, p. V172M was significantly associated with PD, while p. L12M and p. R571C were nominally associated with PD. Burden analysis showed enrichment of ultra-rare variants of PLXNA1 in PD.In addition, one patient carried a variant in the neighboring amino acid, and diaplayed clinical characteristics similar to those of the patient in the original study. Our study explored the rare variant of PLXNA1 in PD in the Asian population and paved the way for future research.

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The tRNA-derived fragments (tRFs) are small non-coding RNAs containing fewer than 50 nucleotides that area bundant in humans and have various biological functions. However, their roles and mechanisms in melanogenesis are unclear. In this study, we firstly investigated the change in the expression profile of tRFs in melanocytes after UVB irradiation through tsRNA sequencing. UVB-induced melanogenesis led to the upregulation of 119 tRFs and the down regulation of 103 tRFs. Among the tRFs with greater than 1.5-fold change in expression level, tRF-Ser-AGA-002 (tRF-5022a) was significantly upregulated after UVB irradiation. Overexpression of tRF-5022a in melanocytes significantly increased melanin production and upregulated melanogenesis-related genes. Ontheotherhand, inhibitionoftRF-5022aexpressionhadthe opposite effects. CCDC88A, which regulates the phosphatidylinositol 3-kinase (PI3K) /protein kinase B (Akt) signaling pathway, was significantly downregulated in the tRF-5022a-overexpressing melanocytes. Collectively, our data indicates that the expression profile of tRFs was significantly altered in melanocytes after UVB irradiation, and tRF-5022a is involved in regulating melanogenesis.

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DNA methylation, as a key epigenetic modification affecting gene expression, is catalyzed by DNA methyl-transferase (DNMT).DNMT1, DNMT3A, and DNMT3B are the main members of the DNMT family, which have been found to regulate many biological processes. Aberrant DNA methylation in the promoter of tumor suppressor genes is considered a biomarker in cancer. However, no evidence of regulatory mechanisms and pathways relating to DNMTs and immune abnormalities has been provided. Herein, we reported a bioinformatic analysis to explore the effect of DNMTs in pan caner. DNMTs are associated with different expressions and poor prognosis in various cancers, especially in LIHC. In addition, we found that DNMTs expression-correlated genes are involved in pathways related to the cell cycle and epithelial-mesenchymal transition (EMT). DNMTs is significantly related to the expression of major immune checkpoints. Furthermore, DNMTs probably engaged in the process of drug resistance. A risk score based on the expression of DNMTs in LIHC was constructed and found that high risk patients have a poor prognosis. Taken together, our study comprehensively analyzed the DNMT family in pan-cancer and demonstrated that DNMTs could be used as prognosis biomarkers and potential therapeutic targets for the development of drugs.

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Super-enhancers are regions of the mammalian genome comprising multiple enhancers in close proximity and often enriched at cancer genes. Recent studies suggest the emerging roles of super-enhancer-associated genes in underlying the cell identity of neuroblastoma (NB) which rises from neural crest-derived cells and is one of the most common malignant solid tumors in children. Tetratricopeptide repeat domain 8 (TTC8), a member of the BBS family of proteins involved in ciliary transport, is our newly identified super-enhancer-associated gene in NB, and its roles in tumorigenesis remain elusive. Herein, TTC8 expression levels were significantly upregulated in NB and correlated with poor prognosis in patients through integrative analysis of public datasets and microarrays. Further investigations show that TTC8 drives the growth and aggressiveness of NB cell lines both in vitro and in vivo. Mechanistically, TTC8 interacts with Paired-like homeobox 2B (PHOX2B), a master transcription factor in NB, to regulate the nucleocytoplasmic distribution of PHOX2B, thereby activating the MAPK signaling pathway, promoting the occurrence and development of NB. In summary, we find that TTC8 might be a potential target for the treatment of NB, warranting further exploration of its specific mechanisms of action.

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Acute lung injury (ALI) globally afflicts over 3 million individuals every year. It can eventually develop into acute respiratory distress syndrome (ARDS) with a high mortality of up to 40%.To date, ALI has been undertreated in terms of the feeble efficacy of clinical approaches and the lack of proven pharmacological targets. G protein-coupled receptors (GPCRs), the promising targets of modern medicine, participate intensively in the regulation of human physiology and pathophysiology. Among them, Gpr116, which is expressed in alveolar epithelial cells and immune cells, has been reported to play a key role in maintaining alveolar homeostasis and inhibiting inflammation. However, its role in the regulation of neutrophil function and ALI remains to be elucidated. In this study, we demonstrated the inhibitory role of Gpr116 on neutrophil function, which contributes to nonspecific lung injury in mice induced by lipopolysaccharide (LPS).

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Despite recent advances in immunotherapy and targeted therapy, the mortality rate seen in lung cancer (LC) is the highest of all cancer forms. The recent topical coronavirus disease 19 (COVID-19) and its subsequent pandemic have resulted in over 505 million confirmed cases and approximately 6.2 million fatalities as of April 2022, with a staggering 30%-50% mortality rate seen in LC patients with COVID-19. Cancerpatients, inparticular, arehighly vulnerable to COVID-19 infection due to immunosuppression, from both the tumor and its treatments. Here we report the impact of the pandemic to be largely negative on LC treatment and research.

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In the era of advancement, the entire world continues to remain baffled by the increased rate of progression of cancer. There has been an unending search for novel therapeutic targets and prognostic markers to curb the oncogenic scenario. The DEAD-box RNA helicases are a large family of proteins characterized by their evolutionary conserved D-E-A-D (Asp-Glu-Ala-Asp) domain and merit consideration in the oncogenic platform. They perform multidimensional functions in RNA metabolism and also in the pathology of cancers. Their biological role ranges from ribosome biogenesis, RNA unwinding, splicing, modification of secondary and tertiary RNA structures to acting as transcriptional coactivators/repressors of various important oncogenic genes. They also play a crucial role in accelerating oncogenesis by promoting cell proliferation and metastasis. DDX5 (p68) is one of the archetypal members of this family of proteins and has gained a lot of attention due to its oncogenic attribute. It is found to be overexpressed in major cancer types such as colon, brain, breast, and prostate cancer. It exhibits its multifaceted nature by not only coactivating genes implicated in cancers but also mediating crosstalk across major signaling pathways in cancer. Therefore, in this review, we aim to illustrate a comprehensive overview of DEAD-box RNA helicases especially p68 by focusing on their multifaceted roles in different cancers and the various signaling pathways affected by them. Further, we have also briefly discoursed the therapeutic interventional approaches with the DEAD-box RNA helicases as the pharmacological targets for designing inhibitors to pave way for cancer therapy.

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Lysine succinylation (Ksuc) is a novel protein post-translational modification (PTM) wherein a succinyl group modifies a lysine residue. Ksuc leads to significant chemical and structural changes to the modified protein. Recent studies have shown that Ksuc might play an important role in organism physiology and some pathophysiological processes, such as tumorigenesis and metabolic diseases. To provide an understanding of the molecular mechanism and functions of Ksuc in different organisms, we reviewed the current literature about Ksuc, mainly summarizing the research advances in eukaryotes and prokaryotes based on both traditional study methods and site prediction tools. We also discussed inhibitors or activators associated with Ksuc that may contribute to proteomic studies and could be useful in future clinical practice. A deeper understanding of Ksuc may shed new light on life science at the protein level and could lead to novel therapeutic strategies for various diseases.

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MicroRNAs (miRNAs) are small non-coding single-stranded RNAs of about 22 nucleotides in length that act as post-transcriptional regulators of gene expression. Depending on the complementarity between miRNA and target mRNA, cleavage, destabilization, or translational suppression of mRNA occurs within the RISC (RNA-induced silencing complex). As gene expression regulators, miRNAs are involved in a variety of biological functions. Dysregulation of miRNAs and their target genes contribute to the pathophysiology of many diseases, including autoimmune and inflammatory disorders. MiRNAs are also present extracellularly in their stable form in body fluids. Their incorporation into membrane vesicles or protein complexes with Ago2, HDL, or nucleophosmin 1 protects them against RNases. Cell-free miRNAs can be delivered to another cell in vitro and maintain their functional potential. Therefore, miRNAs can be considered mediators of intercellular communication. The remarkable stability of cell-free miRNAs and their accessibility in body fluid makes them potential diagnostic or prognostic biomarkers and potential therapeutic targets. Here we provide an overview of the potential role of circulating miRNAs as biomarkers of disease activity, therapeutic response, or diagnosis in rheumatic diseases. Many circulating miRNAs reflect their involvement in the pathogenesis, while for plenty, their pathogenetic mechanisms remain to be explored. Several miRNAs described as biomarkers were also shown to be of therapeutic potential, and some miRNAs are already tested in clinical trials.

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Circadian rhythm refers to the inherent 24-h cycle oscillation of biochemical, physiological and behavioral functions, which is almost universal in eukaryotes. At least 14 core clock genes have been reported to form multiple chain feedback loops that confer intrinsic circadian rhythmicity onto the molecular clock. Accumulating evidence has shown that the circadian gene dysfunction resulted from single nucleotide polymorphisms (SNPs), deletions, epigenetic modification, and deregulation is strongly associated with cancer risk. In the present review, we describe the composition of circadian rhythm system. We highlight the function and mechanism of clock genes in cancer pathogenesis and progression. Moreover, their potential clinical implications as prognostic biomarkers and therapeutic targets have been addressed.

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Wnt signaling executes an indispensable performance in osteoblast differentiation, bone development, homeostasis, and remodeling. Wnt signals trigger the intracellular Wnt signaling cascade to initiate regulating the implication of b-catenin in the bone environment. Going through the novel discoveries done via high-throughput sequencing technologies on genetic mouse models, we highlighted the significant contribution of Wnt ligands, co-receptors, inhibitors, their related skeletal phenotypes in mouse models and the similar bone disorders clinically observed in human beings. Moreover, the crosstalk between Wnt signaling pathway and BMP, TGF-b, FGF, Hippo, Hedgehog, Notch and PDGF signaling pathways is thoroughly demonstrated to be the underlying gene regulatory network that orchestrates osteoblast differentiation and bone development. We also introspected the significance of Wnt signaling transduction in the reorganization of cellular metabolism by stimulating glycolysis, glutamine catabolism, and fatty acid oxidation in osteoblast-lineage cells that display an important regulatory arbor in the cellular bioenergetics of the bone. Throughout this evaluation, most to date therapeutical approaches towards osteoporosis and other bone maladies found in human beings, are formulated with an aspiration to holistically revamp the present clinical applications involving various monoclonal antibodies therapies that lack specificity, efficacy, and safety into more requisite advanced therapeutics that satisfy these three requirements for further clinical considerations. Conclusively, our review provides comprehensive scientific findings related to the fundamental significance of Wnt signaling cascades in skeletal system and the underlying gene regulatory network with other signaling pathways enlightening researchers with the possibility to further integrate the identified target molecules into therapeutic strategies for skeletal disorders treatment in the clinic.

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Maintaining the balance between eliciting immune responses against foreign proteins and tolerating self-proteins is crucial for maintenance of homeostasis. The functions of programmed death protein 1 (PD-1) and its ligand programmed death ligand 1 (PD-L1) are to inhibit immune responses so that over-reacting immune cells does not cause any damage to its own body cells. However, cancer cells hijack this mechanism to attenuate immune cells functions and create an immunosuppressive environment that fuel their continuous growth and proliferation. Over the past few years' rapid development in cancer immunotherapy has opened a new avenue in cancer treatment. Blockade of PD-1 and PD-L1 has become a potential strategy that rescue the functions of immune cells to fight against cancer with high efficacy. Initially, immune checkpoint monotherapies were not very successful, making breast cancer less immunogenic. Although, recent reports support the presence of tumor infiltrating lymphocytes (TILs) in breast cancer that make it favorable for PD-1/PD-L1 mediated immunotherapy, which is effective in PD-L1 positive patients. Recently, anti-PD-1 (pembrolizumab) and anti-PD-L1 (atezolizumab) gets FDA approval for breast cancer treatment and make PD-1/PD-L1 immunotherapy is meaningful for further research. Likewise, this article gathered understanding of PD-1 and PD-L1 in recent years, their signaling networks, interaction with other molecules, regulations of their expressions and functions in both normal and tumor tissue microenvironments are crucial to find and design therapeutic agents that block this pathway and improve the treatment efficacy. Additionally, authors collected and highlighted most of the important clinical trial reports on monotherapy and combination therapy.

1351

Recent advances in deep sequencing technologies have revealed that, while less than 2% of the human genome is transcribed into mRNA for protein synthesis, over 80% of the genome is transcribed, leading to the production of large amounts of noncoding RNAs (ncRNAs). It has been shown that ncRNAs, especially long non-coding RNAs (lncRNAs), may play crucial regulatory roles in gene expression. As one of the first isolated and reported lncRNAs, H19 has gained much attention due to its essential roles in regulating many physiological and/or pathological processes including embryogenesis, development, tumorigenesis, osteogenesis, and metabolism. Mechanistically, H19 mediates diverse regulatory functions by serving as competing endogenous RNAs (CeRNAs), Igf2/H19 imprinted tandem gene, modular scaffold, cooperating with H19 antisense, and acting directly with other mRNAs or lncRNAs. Here, we summarized the current understanding of H19 in embryogenesis and development, cancer development and progression, mesenchymal stem cell lineage-specific differentiation, and metabolic diseases. We discussed the potential regulatory mechanisms underlying H19's functions in those processes although more in-depth studies are warranted to delineate the exact molecular, cellular, epigenetic, and genomic regulatory mechanisms underlying the physiological and pathological roles of H19.Ultimately, these lines of investigation may lead to the development of novel therapeutics for human diseases by exploiting H19 functions.

1646

Cancer is an abnormal state of cells where they undergo uncontrolled proliferation and produce aggressive malignancies that causes millions of deaths every year. With the new understanding of the molecular mechanism (s) of disease progression, our knowledge about the disease is snowballing, leading to the evolution of many new therapeutic regimes and their successive trials. In the past few decades, various combinations of therapies have been proposed and are presently employed in the treatment of diverse cancers. Targeted drug therapy, immunotherapy, and personalized medicines are now largely being employed, which were not common a few years back. The field of cancer discoveries and therapeutics are evolving fast as cancer type-specific biomarkers are progressively being identified and several types of cancers are nowadays undergoing systematic therapies, extending patients' disease-free survival thereafter. Although growing evidence shows that a systematic and targeted approach could be the future of cancer medicine, chemotherapy remains a largely opted therapeutic option despite its known side effects on the patient's physical and psychological health. Chemotherapeutic agents/pharmaceuticals served a great purpose over the past few decades and have remained the frontline choice for advanced-stage malignancies where surgery and/or radiation therapy cannot be prescribed due to specific reasons. The present report succinctly reviews the existing and contemporary advancements in chemotherapy and assesses the status of the enrolled drugs/pharmaceuticals; it also comprehensively discusses the emerging role of specific/targeted therapeutic strategies that are presently being employed to achieve better clinical success/survival rate in cancer patients.

1248

Severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) causes the complicated disease COVID-19. Clinicians are continuously facing huge problems in the treatment of patients, as COVID-19-specific drugs are not available, hence the principle of drug repurposing serves as a one-and-only hope. Globally, the repurposing of many drugs is underway; few of them are already approved by the regulatory bodies for their clinical use and most of them are in different phases of clinical trials. Here in this review, our main aim is to discuss in detail the up-to-date information on the target-based pharmacological classification of repurposed drugs, the potential mechanism of actions, and the current clinical trial status of various drugs which are under repurposing since early 2020.At last, we briefly proposed the probable pharmacological and therapeutic drug targets that may be preferred as a futuristic drug discovery approach in the development of effective medicines.

1352

DNA is highly vulnerable to spontaneous and environmental timely damage in living cells. DNA damage may cause genetic instability and increase cancer risk if the damages are not repaired timely and efficiently. Human cells possess several DNA damage response (DDR) mechanisms to protect the integrity of their genome. Clarification of the mechanisms underlying the DNA damage response following lethal damage will facilitate the identification of therapeutic targets for cancers. Histone post-translational modifications (PTMs) have been indicated to play different roles in the repair of DNA damage. In this context, histone PTMs regulate recruitment of downstream effectors, and facilitate appropriate repair response. This review outlines the current understanding of different histone PTMs in response to DNA damage repair, besides, enumerates the role of new type PTMs such as histone succinylation and crotonylation in regulating DNA damage repair processes.

1284

Globally, cervical cancer (CxCa) ranks 4th common cancer among females and led to 569,847 incidences and 311,365 deaths in 2018.80% of CxCa cases occur due to persistent infection with a high-risk subtype of human papillomavirus (HPV-16 and 18). Smoking, high parity, and co-infection with type 2 herpes simplex or HIV are other known risk factors for CxCa. Major histological subtypes are squamous (70%) and adenocarcinoma (25%).Presently, concurrent radiation plus cisplatin (CDDP) -based chemotherapy is the standard treatment for CxCa patients. However, CDDP resistance and toxic side effects limit its efficacy, leading to a poorer response rate and an expected overall survival ranging from 10 to 17.5 months. Reduced drug uptake, increased DNA damage repair, increased CDDP inactivation, and overexpressed Bcl-2 or caspase inhibition, are primarily accountable mechanisms for CDDP resistance and improving CDDP's efficacy remains the major challenge. Poly (ADP-ribosyl) polymerase-1, an effective mediator of nucleotide excision repair pathway, is involved in DNA repair as well as maintaining genomic stability and is significantly expressed in malignant lymphomas, hepatocellular-, cervical-and colorectal carcinoma, which has been approved effective in maintenance therapy and may serve as an effective target to enhance CDDP sensitivity in CxCa. Here, we summarize the etiology and epidemiology of and treatment for CxCa, the mechanism responsible for chemotherapy resistance, PARP inhibitor as a possible therapy for CxCa, and other possible chemotherapeutic options for CxCa treatment.

1340

microRNAs (miRNAs) are a class of non-coding functional small RNA composed of 21-23 nucleotides, having multiple associations with liver fibrosis. Fibrosis-associated miRNAs are roughly classified into pro-fibrosis or anti-fibrosis types. The former is capable of activating hepatic stellate cells (HSCs) by modulating pro-fibrotic signaling pathways, mainly including TGF-b/SMAD, WNT/β -catenin, and Hedgehog; the latter is responsible for maintenance of the quiescent phenotype of normal HSCs, phenotypic reversion of activated HSCs (aHSCs), inhibition of HSCs proliferation and suppression of the extracellular matrix-associated gene expression. Moreover, several miRNAs are involved in regulation of liver fibrosis via alternative mechanisms, such as interacting between hepatocytes and other liver cells via exosomes and increasing autophagy of aHSCs. Thus, understanding the role of these miRNAs may provide new avenues for the development of novel interventions against hepatic fibrosis.

1384

Detrimental impacts of fluoride have become a global concern for several decades. Despite its beneficial role which is restricted only in skeletal tissues, deleterious effects are also observed in soft tissues and systems. The generation of enhanced oxidative stress is the commencement of excess fluoride exposure which may lead to cell death. Fluoride causes cell death through autophagy via Beclin 1 and mTOR signaling pathways. Beside these, several organ specific anomalies through different signaling pathways have been documented. Mitochondrial dysfunction, DNA damage, autophagy and apoptosis are the damaging outcomes in case of hepatic disorders. Urinary concentration defects and cell cycle arrest have been reported in renal tissues. Abnormal immune response has been characterized in the cardiac system. Cognitive dysfunction, neurodegenerative condition and learning impairment have also been observed. Altered steroidogenesis, gametogenic abnormalities, epigenetic alterations and birth defect are the major reprotoxic conclusions. Abnormal immune responses, altered immunogenic proliferation, differentiation as well as altered ratio of immune cells are well-defined anomalies in the immune system. Though the mechanistic approach of fluoride toxicity in physiological systems is common, it follows different signaling cascades. This review emphasizes diverse signaling pathways which are the targets of overexposed fluoride.

1298

All cells release extracellular vesicles (EVs) as part of their normal physiology. As one of the subtypes, exosomes (EXOs) have an average size range of approximately 40 nm e160 nm in diameter. Benefiting from their inherent immunogenicity and biocompatibility, the utility of autologous EXOs has the potential for both disease diagnosis/treatment. EXOs are generally employed as "bioscaffolds" and the whole diagnostic and therapeutic effects are mainly ascribed to exogenous cargos on the EXOs, such as proteins, nucleic acids, and chemotherapeutic agents and fluorophores delivered into specific cells or tissues. Surface engineering of EXOs for cargo loadings is one of the prerequisites for EXO-mediated diagnosis/treatment. After revisiting EXO-mediated diagnosis/treatment, the most popular strategies to directly undertake loadings of exogenous cargos on EXOs include genetic and chemical engineering. Generally, genetically-engineered EXOs can be merely produced by living organisms and intrinsically face some drawbacks. However, chemical methodologies for engineered EXOs diversify cargos and extend the functions of EXOs in the diagnosis/treatment. In this review, we would like to elucidate different chemical advances on the molecular level of EXOs along with the critical design required for diagnosis/treatment. Besides, the prospects of chemical engineering on the EXOs were critically addressed. Nevertheless, the superiority of EXO-mediated diagnosis/treatment via chemical engineering remains a challenge in clinical translation and trials. Furthermore, more chemical crosslinking on the EXOs is expected to be explored. Despite substantial claims in the literature, there is currently no review to exclusively summarize the chemical engineering to EXOs for diagnosis/treatment. We envision chemical engineering of EXOs will encourage more scientists to explore more novel technologies for a wider range of biomedical applications and accelerate the successful translation of EXO-based drug "bioscaffolds" from bench to bedside.

1421

Pancreatic cancer (PC) is an aggressive malignant tumor with low rate of surgical resection and poor prognosis. Transforming growth factor-β (TGF-b) is a cytokine that has both protumor and antitumor activities, depending on tumor microenvironment. The interaction between TGF-b signaling and the tumor microenvironment in PC is complex. Here, we reviewed the role of TGF-b in the tumor microenvironment of PC, highlighting producers of TGF-b and TGF-b responders in the tumor microenvironment of PC.

1311

Metabolic cardiovascular diseases have become a global health concern, and some of their risk factors are linked to several metabolic disorders. They are the leading causes of death in developing countries. Adipose tissues secrete a variety of adipokines that participate in regulating metabolism and various pathophysiological processes. Adiponectin is the most abundant pleiotropic adipokine and can increase insulin sensitivity, improve atherosclerosis, have anti-inflammatory properties, and exert a cardioprotective effect. Low adiponectin concentrations are correlated with myocardial infarction, coronary atherosclerotic heart disease, hypertrophy, hypertension, and other metabolic cardiovascular dysfunctions. However, the relationship between adiponectin and cardiovascular diseases is complex, and the specific mechanism of action is not fully understood. Our summary and analysis of these issues are expected to contribute to future treatment options.

1356

Placental growth factor (PlGF) is a glycosylated dimeric protein that is homologous to vascular endothelial growth factor (VEGF). PlGF expression is upregulated in patients with bronchial asthma, suggesting that it plays a role in the pathogenesis of asthma. Bronchial asthma is characterized by chronic airway inflammation and airway hyperresponsiveness (AHR).After recurrent asthma attacks, pulmonary fibrosis develops and leads to airway remodeling and a further decline in lung function. In this review, we focused on the pivotal role of PlGF in chronic airway inflammation, AHR, and airway remodeling during bronchial asthma. Furthermore, we summarized data showing that PlGF may be a potential therapeutic target in bronchial asthma.

1424

Inflammatory bowel disease (IBD) is a chronic relapsing gastrointestinal disorder, while the treatment effect is not satisfactory. Immune responsive gene 1 (IRG1) is a highly expressed gene in macrophage in response to inflammatory response and catalyzes the production of itaconate. Studies have reported that IRG1/itaconate has a significant antioxidant effect. This study aimed to investigate the effect and mechanism of IRG1/itaconate on dextran sulfate sodium (DSS)-induced colitis in vivo and in vitro. In vivo experiments, we found IRG1/itaconate exerted protective effects against acute colitis by increasing mice weight, the length of colon, reducing disease activity index and colonic inflammation. Meanwhile, IRG1 deletion aggravated the macrophages/CD4⁺/CD8⁺ T-cell accumulation, and increased the release of interleukin (IL)-1β, tumor necrosis factor-a (TNF-α), IL-6, the activation of nuclear factor-κB (NF-κB)/mitogenactivated protein kinase (MAPK) signaling pathway, and gasdermin D (GSDMD) mediated pyroptosis. Four-octyl itaconate (4-OI), a derivative of itaconate, attenuated these changes, therefore relieved DSS-induced colitis. In vitro experiment, we found 4-OI inhibited the reactive oxygen species production, thereby inhibiting the activation of MAPK/NF-κB signaling pathway in RAW264.7 and murine bone-marrow-derived macrophages. Simultaneously,we found 4-OI inhibited caspase1/GSDMD-mediated pyroptosis to reduce the release of cytokines. Finally, we found anti-TNF-α agent reduced the severity of DSS-induced colitis and inhibited gasdermin E (GSDME)-mediated pyroptosis in vivo. Meanwhile, our study revealed that 4-OI inhibited caspase3/GSDME-mediated pyroptosis induced by TNF-α in vitro. Taken together, IRG1/itaconate exerted a protective role in DSS-induced colitis by inhibiting inflammatory response and GSDMD/GSDME-mediated pyroptosis, which could be a promising candidate for IBD therapy.

1435

Glaucoma is the leading cause of irreversible blindness worldwide. In the pathogenesis of glaucoma, activated microglia can lead to retinal ganglion cells (RGCs) apoptosis and death, however, the molecular mechanisms remain largely unknown. We demonstrate that phospholipid scramblase 1 (PLSCR1) is a key regulator promoting RGCs apoptosis and their clearance by microglia. As evidenced in retinal progenitor cells and RGCs of the acute ocular hypertension (AOH) mouse model, overexpressed PLSCR1 induced its translocation from the nucleus to the cytoplasm and cytomembrane, as well as elevated phosphatidylserine exposure and reactive oxygen species generation with subsequent RGCs apoptosis and death. These damages were effectively attenuated by PLSCR1 inhibition. In the AOH model, PLSCR1 led to an increase in M1 type microglia activation and retinal neuroinflammation. Upregulation of PLSCR1 resulted in strongly elevated phagocytosis of apoptotic RGCs by activated microglia. Taken together, our study provides important insights linking activated microglia to RGCs death in the glaucoma pathogenesis and other RGC-related neurodegenerative diseases.

1304

Progranulin (PGRN) is a multifunctional growth factor involved in many physiological processes and disease states. The apparent protective role of PGRN and the importance of chondrocyte autophagic function in the progression of osteoarthritis (OA) led us to investigate the role of PGRN in the regulation of chondrocyte autophagy. PGRN knockout chondrocytes exhibited a deficient autophagic response with limited induction following rapamycin, serum starvation, and IL-1b-induced autophagy. PGRN-mediated anabolism and suppression of IL-1b-induced catabolism were largely abrogated in the presence of the BafA1 autophagy inhibitor. Mechanistically, during the process of OA, PGRN and the ATG5eATG12 conjugate form a protein complex; PGRN regulates autophagy in chondrocytes and OA through, at least partially, the interactions between PGRN and the ATG5eATG12 conjugate. Furthermore, the ATG5eATG12 conjugate is critical for cell proliferation and apoptosis. Knockdown or knockout of ATG5 reduces the expression of ATG5eATG12 conjugate and inhibits the chondroprotective effect of PGRN on anabolism and catabolism. Overexpression of PGRN partially reversed this effect. In brief, the PGRN-mediated regulation of chondrocyte autophagy plays a key role in the chondroprotective role of PGRN in OA. Such studies provide new insights into the pathogenesis of OA and PGRN-associated autophagy in chondrocyte homeostasis.

1422

Non-alcoholic steatohepatitis (NASH), a progressive form of non-alcoholic fatty liver disease (NAFLD), is characterised by chronic liver inflammation, which can further progress into complications such as liver cirrhosis and NASH-associated hepatocellular carcinoma (HCC) and therefore has become a growing health problem worldwide. The type I interferon (IFN) signaling pathway plays a pivotal role in chronic inflammation; however, the molecular mechanisms underlying NAFLD/NASH from the perspective of innate immune response has not yet been fully explored. In this study, we elucidated the mechanisms of how innate immune response modulates NAFLD/NASH pathogenesis, and demonstrated that hepatocyte nuclear factor-1alpha (HNF1A) was suppressed and the type I IFN production pathway was activated in liver tissues of patients with NAFLD/NASH. Further experiments suggested that HNF1A negatively regulates the TBK1-IRF3 signaling pathway by promoting autophagic degradation of phosphorylated-TBK1, which constrains IFN production, thereby inhibiting the activation of type I IFN signaling. Mechanistically, HNF1A interacts with the phagophore membrane protein LC3 through its LIR-docking sites, and mutations of LIRs (LIR2, LIR3, LIR4, and LIRs) block the HNF1A-LC3 interaction. In addition, HNF1A was identified not only as a novel autophagic cargo receptor but also to specifically induce K33-linked ubiquitin chains on TBK1 at Lys670, thereby resulting in autophagic degradation of TBK1.Collectively, our study illustrates the crucial function of the HNF1A-TBK1 signaling axis in NAFLD/NASH pathogenesis via crosstalk between autophagy and innate immunity.

1361

Extracellular vesicles (EVs) derived from mesenchymal stem cells (MSCs) have emerged as a new mode of intercellular crosstalk and are responsible for many of the therapeutic effects of MSCs. To promote the application of MSC-EVs, recent studies have focused on the manipulation of MSCs to improve the production of EVs and EV-mediated activities. The current paper details an optimization method using non-invasive low-intensity pulsed ultrasound (LIPUS) as the stimulation for improving oral MSC-EV production and effectiveness. Stem cells from apical papilla (SCAP), a type of oral mesenchymal stem cell, displayed intensity-dependent pro-osteogenic and anti-inflammatory responses to LIPUS without significant cytotoxicity or apoptosis. The stimuli increased the secretion of EVs by promoting the expression of neutral sphingomyelinases in SCAP. In addition, EVs from LIPUS-induced SCAP exhibited stronger efficacy in promoting the osteogenic differentiation and anti-inflammation of periodontal ligament cells in vitro and alleviating oral inflammatory bone loss in vivo.In addition, LIPUS stimulation affected the physical characteristics and miRNA cargo of SCAP-EVs. Further investigations indicated that miR-935 is an important mediator of the pro-osteogenic and anti-inflammatory capabilities of LIPUS-induced SCAP-EVs. Taken together, these findings demonstrate that LIPUS is a simple and effective physical method to optimize SCAP-EV production and efficacy.

1567

More than 50% of prostate cancer (PCa) patients have bone metastasis with osteoblastic lesions. MiR-18a-5p is associated with the development and metastasis of PCa, but it remains unclear whether it is involved in osteoblastic lesions. We first found that miR-18a-5p was highly expressed in the bone microenvironment of patients with PCa bone metastases. To address how miR-18a-5p affects PCa osteoblastic lesions, antagonizing miR-18a-5p in PCa cells or pre-osteoblasts inhibited osteoblast differentiation in vitro.Moreover, injection of PCa cells with miR-18a-5p inhibition improved bone biomechanical properties and bone mineral mass in vivo.Furthermore, miR-18a-5p was transferred to osteoblasts by exosomes derived from PCa cells and targeted the Hist1h2bc gene, resulting in Ctnnb1 up-regulation in the Wnt/β -catenin signaling pathway. Translationally, antagomir-18a-5p significantly improved bone biomechanical properties and alleviated sclerotic lesions from osteoblastic metastases in BALB/c nude mice. These data suggest that inhibition of exosome-delivered miR-18a-5p ameliorates PCa-induced osteoblastic lesions.

1347

Cancer cells tend to develop resistance to chemotherapy and enhance aggressiveness. A counterintuitive approach is to tame aggressiveness by an agent that acts opposite to chemotherapeutic agents. Based on this strategy, induced tumor-suppressing cells (iTSCs) have been generated from tumor cells and mesenchymal stem cells. Here, we examined the possibility of generating iTSCs from lymphocytes by activating PKA signaling for suppressing the progression of osteosarcoma (OS).While lymphocyte-derived CM did not present anti-tumor capabilities, the activation of PKA converted them into iTSCs. Inhibiting PKA conversely generated tumor-promotive secretomes. In a mouse model, PKA-activated CM suppressed tumor-induced bone destruction. Proteomics analysis revealed that moesin (MSN) and calreticulin (Calr), which are highly expressed intracellular proteins in many cancers, were enriched in PKA-activated CM, and they acted as extracellular tumor suppressors through CD44, CD47, and CD91.The study presented a unique option for cancer treatment by generating iTSCs that secret tumor-suppressive proteins such as MSN and Calr. We envision that identifying these tumor suppressors and predicting their binding partners such as CD44, which is an FDA-approved oncogenic target to be inhibited, may contribute to developing targeted protein therapy.

1288

The high risk of postoperative mortality in lung adenocarcinoma (LUAD) patients is principally driven by cancer recurrence and low response rates to adjuvant treatment. Here, A combined cohort containing 1,026 stage IeIII patients was divided into the learning (n = 678) and validation datasets (n = 348). The former was used to establish a 16-mRNA risk signature for recurrence prediction with multiple statistical algorithms, which was verified in the validation set. Univariate and multivariate analyses confirmed it as an independent indicator for both recurrence-free survival (RFS) and overall survival (OS).Distinct molecular characteristics between the two groups including genomic alterations, and hallmark pathways were comprehensively analyzed. Remarkably, the classifier was tightly linked to immune infiltrations, highlighting the critical role of immune surveillance in prolonging survival for LUAD. Moreover, the classifier was a valuable predictor for therapeutic responses in patients, and the low-risk group was more likely to yield clinical benefits from immunotherapy. A transcription factor regulatory proteineprotein interaction network (TF-PPI-network) was constructed via weighted gene co-expression network analysis (WGCNA) concerning the hub genes of the signature. The constructed multidimensional nomogram dramatically increased the predictive accuracy. Therefore, our signature provides a forceful basis for individualized LUAD management with promising potential implications.

1347

Vaccination by inactivated vaccine is an effective strategy to prevent the COVID-19 pandemic. However, the detailed molecular immune response at single-cell level is poorly understood. In this study, we systematically delineated the landscape of the pre-and post-vaccination single-cell transcriptome, TCR (T cell antigen receptor) and BCR (B cell antigen receptor) expression profile of vaccinated candidates. The bulk TCR sequencing analysis of COVID-19 patients was also performed. Enrichment of a clonal CD8⁺T cell cluster expressing specific TCR was identified in both vaccination candidates and COVID-19 patients. These clonal CD8⁺T cells showed high expression of cytotoxicity, phagosome and antigen presentation related genes. The cellecell interaction analysis revealed that monocytes and dendritic cells could interact with these cells and initiate phagocytosis via ICAM1-ITGAM and ITGB2 signaling. Together, our study systematically deciphered the detailed immunological response during SARS-CoV-2 vaccination and infection. It may facilitate understanding the immune response and the T-cell therapy against COVID-19.

1463

Ovarian cancer (OC) is one of the most lethal malignancies of the female reproductive system. OC patients are usually diagnosed at advanced stages due to the lack of early diagnosis. The standard treatment for OC includes a combination of debulking surgery and platinum-taxane chemotherapy, while several targeted therapies have recently been approved for maintenance treatment. The vast majority of OC patients relapse with chemoresistant tumors after an initial response. Thus, there is an unmet clinical need to develop new therapeutic agents to overcome the chemoresistance of OC. The anti-parasite agent niclosamide (NA) has been repurposed as an anti-cancer agent and exerts potent anti-cancer activities in human cancers including OC. Here, we investigated whether NA could be repurposed as a therapeutic agent to overcome cisplatin-resistant (CR) in human OC cells. To this end, we first established two CR lines SKOV3CR and OVCAR8CR that exhibit the essential biological characteristics of cisplatin resistance in human cancer. We showed that NA inhibited cell proliferation, suppressed cell migration, and induced cell apoptosis in both CR lines at a low micromole range. Mechanistically, NA inhibited multiple cancer-related pathways including AP1, ELK/SRF, HIF1, and TCF/LEF, in SKOV3CR and OVCAR8CR cells. NA was further shown to effectively inhibit xenograft tumor growth of SKOV3CR cells. Collectively, our findings strongly suggest that NA may be repurposed as an efficacious agent to combat cisplatin resistance in chemoresistant human OC, and further clinical trials are highly warranted.

1360

How PD-L1 expression is regulated in cancer is poorly understood. Here, we report that the ATP-binding activity of ERBB3 pseudokinase regulates PD-L1 gene expression in colorectal cancers (CRCs).ERBB3 is one of the four members of the EGF receptor family, all with protein tyrosine kinase domains. ERBB3 is a pseudokinase with a high binding affinity to ATP. We showed that ERBB3 ATP-binding inactivation mutant reduces tumorigenicity in genetically engineered mouse models and impairs xenograft tumor growth of CRC cell lines. The ERBB3 ATP-binding mutant cells dramatically reduce IFN-g-induced PD-L1 expression. Mechanistically, ERBB3 regulates IFN-g-induced PD-L1 expression through the IRS1-PI3K-PDK1-RSK-CREB signaling axis. CREB is the transcription factor that regulates PD-L1 gene expression in CRC cells. Knockin of a tumor-derived ERBB3 mutation located in the kinase domain sensitizes mouse colon cancers to anti-PD1 antibody therapy, suggesting that ERBB3 mutations could be predictive biomarkers for tumors amenable to immune checkpoint therapy.

1439

Osteoarthritis (OA) is a chronic debilitating joint disease, characterized by degeneration of the cartilage and loss of the cartilage matrix, and it is clinically manifested as joint pain. Osteopontin (OPN) is a glycoprotein that is abnormally expressed in the bone and cartilage tissues and plays a vital role in various pathological processes such as the osteoarthritic inflammatory response and endochondral ossification. The focus of our study is to investigate the therapeutic potential and specific role of OPN in OA. Using morphological comparisons, we found that the cartilage was severely worn-out and there was a significant loss of the cartilage matrix in OA. OPN, CD44, and hyaluronic acid (HA) synthase 1 (HAS1) were highly expressed, and the anabolism of HA was significantly higher in the OA chondrocytes than in the control chondrocytes. Additionally, we treated the OA chondrocytes with small interfering RNA (siRNA) targeting OPN, recombinant human OPN (rhOPN), and a combination of rhOPN and anti-CD44 antibodies. Furthermore, in vivo experiments were performed in mice. We found that OPN upregulated the expression of downstream HAS1 and increased the anabolism of HA through CD44 protein expression in OA mice compared with those in control mice. Moreover, intra-articular injection of OPN in mice with OA significantly inhibited OA progression. In summary, OPN initiates an intracellular cascade via CD44 which results in an anabolic increase in HA levels, thereby inhibiting OA progression. Therefore, OPN is a promising therapeutic agent in precision treatment of OA.

1355