第9卷，第2期

Dear Editor,Triple-negative breast cancer (TNBC) is an aggressive breast cancer subtype enriched in cancer stem cells (CSCs), which is characterized by high malignancy and drug resistance. The PI3K/Akt signaling cascade is frequently hyperactivated in cancers. Distinct and non-redundant functions of the three Akt isoforms in tumorigenesis, however, have not been studied extensively. It is noteworthy that despite a few isoform-specific substrates have been identified for Akt1 and Akt2, an Akt3-specific substrate is yet to be discovered. We have previously shown that overexpression of Akt3, but not Akt1 or Akt2, plays an essential role in TNBC growth and therapeutic resistance of breast cancer. However, the role of Akt3 in TNBC CSCs remains unclear.

1238

In healthy humans, ammonia circulates in the bloodstream at a concentration around 11-32 mmol/L. Whether ammonia plays any physiological role has been unknown until a recent report published in Nature Metabolism. Ammonia, long thought to be a toxic waste product of amino acid metabolism, needs to be excreted from the human body as urea. Intriguingly, in this paper, Cheng et al for the first time reported that ammonia is not a waste product; rather, it is an unprecedented signaling molecule tying glucose and glutamine to lipid production in human cells. This study revealed that ammonia released from glutamine triggers a cascade of cellular processes leading to the activation of lipogenesis machinery for lipid production. This discovery has major implications for developing treatments for human diseases such as cancers and metabolic syndromes, as dysregulated metabolism is a hallmark of these diseases.

1317

Salt-inducible kinase 2 (SIK2) is a member of the AMP-activated serine/threonine kinase family. It has been reported that inhibition of SIK2 can enhance the cytotoxicity of paclitaxel, promote premitotic apoptosis, and lead to cell cycle arrest in the metaphase. Thus, targeting SIK2 may be a therapeutic strategy for cancers drug and radiotherapy resistance. Mitotic catastrophe is a type of abnormal mitosis leading to cell death characterized by the multipolar spindle and multinucleation, which was first discovered during an ionizing radiation (IR) -induced cell damage. However, the mechanism of mitotic catastrophe is not well understood. The present study aimed to assess the effect of the knockdown of SIK2 on IR-induced mitotic catastrophe.

1383

Long noncoding RNAs (lncRNAs) are known to be involved in transcriptional regulation and their deregulation is associated with the development of human diseases such as cancer. LncRNA can directly bind to purine-rich doublestranded DNA sequences through Hoogsteen base pairing to form an RNA-DNA triplex motifs and regulate gene expression. However, its regulatory mechanisms and functions in tumors remain unclear. Here, we report that the LMNTD2 antisense RNA 1 (LMNTD2-AS1, also known as AP006284. 1) is highly expressed in prostate cancer (PCa) and positively correlated with the expression of its adjacent coding gene N-terminal Ras-association domain family 7 (RASSF7). Mechanistically, AP006284. 1 tethers to the RASSF7promoterviaRNA-DNAtriplexesincis-acting manner, which enhances chromatin accessibility and recruits the transcription factor complex GNL3/SFPQ to activate the expression of RASSF7, a repressor of the Hippo signaling pathway. Consistently, overexpression of either AP006284. 1 or RASSF7 inactivated the Hippo signaling and malignant proliferation of PCa cells.

1322

Amyotrophic lateral sclerosis (ALS) is a fatal motor neuron disease typically leading to death within 5 years from symptom onset. ALS familial forms are associated with mutations in several genes, including Superoxide Dismutase 1 (SOD1) and Fused in Sarcoma (FUS). Although genes linked to ALS participate in disparate biological processes, ALS genetic variants largely trigger shared pathogenic events such as oxidative stress, protein aggregation and defects in RNA processing. Moreover, ALS patients show systemic hypermetabolism that leads to increased energy expenditure at rest and thus weight loss during the disease course. ALS hypermetabolic phenotype and weight loss have been extensively characterized in mice bearing the G93A substitution in SOD1 protein (SOD1-G93A), which exhibit skeletal-muscle metabolic reprogramming before disease onset.

1830

Lung metastasis is the primary cause of death in osteosarcoma (OS) patients. A better understanding of the molecular mechanisms underlying OS tumorigenesis and metastasis is urgently needed to identify therapeutic targets. Microtubule actin crosslinking factor 1 (MACF1), which belongs to the spectraplakin family of cytoskeletal crosslinking proteins, is critical for cell migration and polarization due to its regulation of the cytoskeleton. Recently, MACF1 was indicated to be involved in the metastatic invasion of some human cancers, but the function of MACF1 in OS is still unclear.

1262

Mutations in the Ras oncogene are the most frequently cancer alterations, occurring in more than 30% of all human cancers. The failures in the development of successful clinical inhibitors against Ras have made Ras as a "undruggable" target. The major reason is lack of a systematic understanding of oncogenic cooperation between Ras activation and mutation of related tumor suppressor genes. The genetic techniques available in the Drosophila melanogaster allow analysis of the behavior of cells with distinct mutations, making this the ideal model organism to dissect oncogenic cooperation induced tumorigenesis. The exact samemutation, e. g., Ras^V12, was mimicked in the Drosophila, which enables us to perform large scale genetic screens, aiming to unearth novel tumor suppressors that can synergistically enhance Ras^V12-related tumor growth.

1245

1337

SETD2 is the main transferase for the trimethylation of histone H3 at lysine 36 (H3K36me3) in mammals. SETD2 plays important role in repairing DNA double strand breaks and maintaining chromatin integrity. In renal carcinoma, SETD2 deficiency caused DNA replication fork instability and DNA damage. The absence of SETD2 was also reported to have strong tumor-promoting effects in lung adenocarcinomas (LUAD). However, the other roles of SETD2 remain poorly understood in LUAD. In this study, we utilized comprehensive omics-data analysis to determine that SETD2 was associated with DNA damage and immunerelated signals in LUAD.SETD2 knockdown induced DNA damage and cGAS activation in LUAD cells, and reduced the number of cells at the G1 phase. Moreover, SETD2 deficiency was conducive to mutation burden, immune cellinfiltration, and immunotherapy responses. High SETD2 expression was associated with highradio-curability. These findings suggest that SETD2 may be a promising biomarker of therapeutic responses for LUAD patients, and offer novel insights into immunotherapy and radiotherapy.

1328

Type II diabetes and obesity are two of the most prevalent metabolic disorders effecting a huge population throughout the world. Research over the last decade has unequivocally established considerable molecular links between them and hence they are often described in conjugation as 'diabesity'. The hallmarks of type II diabetes are primarily reduced insulin sensitivity, progressive insulin resistance and consequent hyperinsulinemia. Whereas, hyperinsulinemia promotes a plethora of fat synthesis from excess circulating carbohydrate and hyperactive fat storage mechanisms, ultimately inducing hepatic steatosis, myosteatosis and pancreatic steatosis. This in turn, aggravates the insulin sensitivity further, inflicting more insulin resistance and even more hyperinsulinemia. Needless to mention that the vicious process in turn, promotes uncontrolled weight gain and many secondary metabolic disorders. The prevalent anti-diabetic and anti-obesity medications comes with several limitations ranging from inefficiency to adverse side effects. Here, we report an efficient strategy of repositioning previously approved drugs with novel indication in the context of diabesity by investigating deregulated signalling axes affecting patients with both the disorders. Our approach relies extensively on deciphering the strength of gene association in various interactomes, as it is known that within networks, genes linked to similar disease phenotypes tend to be functionally similar and remain proximal to each other. Moreover, the potential drug targets associated to a disease pathway also cluster proximal to the disease pathways.

1306

The signal adaptor myeloid differentiation primary response 88 (MyD88) of Toll-like receptor (TLR) signaling is universally expressed in immune cells and non-immune cells, and myeloid cells play a significant role in modulating colitis. Macrophages are myeloid lineage cells which is important for maintaining intestinal homeostasis in inflammation. And macrophages can recognize invading pathogens through pattern recognition receptors (PRRs), such as TLRs and the nucleotide oligomerization domain (NOD) -like receptor family, pyrin domain-containing 3 (NLRP3), and quickly infiltrate the injured site leading to inflammation. MyD88 expression in myeloid cells can rescue the intestinal injury induced by dextran sulfate sodium (DSS) in murine models. However, the mechanism whereby MyD88 works in myeloid cells and influence the progress of the disease remain elusive. In this study, we used DSS induced colitis mouse model and found Lysm-cre-MyD88^fl/fl (MyD88^ΔMΦ) mice were more susceptible to colitis. The deficiency of MyD88 leads to up-regulation of S100A8, which activate NLRP3 inflammasome and their associated pyroptosis in intestinal epithelial cells in a RAGE dependent manner. Thus, MyD88 signaling in macrophages, which is necessary to maintain intestinal homeostasis, crucially prevents the development of colitis. Our results may shed new lights on potential targets that can be applied in colitis therapy.

1275

Dysregulation of the Hippo pathway has been frequently identified in various human cancers. The Hippo pathway is a highly complex pathway involving multiple types of proteins, and the activation of YAP by LATS kinase is the final effector step in the transcription process. In this study, we linked the roles of the multifunctional adapter proteins β-arrestins (barrestin 1 and 2) in cooperation with other signaling pathways such as GPCR and Wnt to essential cellular functions involved in carcinogenesis, including the regulation of cell proliferation, migration, and differentiation as well as stem cell properties. Although β-arrestin 1 and 2 have high structural similarities, they have different roles in carcinogenesis as β-arrestin 1 aids cancer cell survival and metastasis and β-arrestin 2 inhibits tumor growth. In the Hippo signaling pathway, β-arrestins function as scaffold proteins that mediate the phosphorylation of key molecules, and their association with human cancers is a major research topic. In this study, we demonstrated that β-arrestin 2 inhibits YAP activation through the formation of the β-arrestin 2-LATS-YAP trimeric complex, which results in the promotion of the kinase activity of LATS in cancer cell lines and patient-derived colon cancer organoids.

1807

Metabolic reprogramming is one of the hallmarks of cancer. Ketone bodies behave as alternative fuel for cancer cells to support survival and proliferation. 3-Oxoacid CoA-transferase 1 (OXCT1) is a key enzyme that catalyzes the first and rate-limiting step of ketolysis. Recently, several studies have revealed the significance of OXCT1 in cancer development, though the underlying mechanisms remain largely unknown. In this study, we revealed a novel regulatory mechanism for tumorigenesis that OXCT1 regulated SREBP1-TRIM21-p65 axis through ketone body homeostasis in non-small cell lung cancer (NSCLC). In terms of mechanism, we found that OXCT1 could activate NF-κB signaling pathway by suppressing transcriptional activity of the sterol regulatory element binding protein 1 (SREBP1). As a transcription factor, SREBP1 could bind to the promoter of E3 ubiquitin ligase TRIM21, which mediated the ubiquitination of p65. Furthermore, we demonstrated that OXCT1 could maintain the homeostasis of β-hydroxybutyrate (β-HB), which acted as a signaling metabolite to activate SREBP1. Thus, β-HB connected OXCT1 with SREBP1 to activate NF-κB signaling pathway and promoted tumor initiation and progression. Taken together, these findings highlight a previously unappreciated mechanism for activation of NF-κB signaling by OXCT1 and ketone body, and demonstrate that targeting OXCT1 can inhibit NSCLC tumorigenesis.

1226

Osteosarcoma (OS) is the most common histological form of primary bone cancer in childhood cancer and young adults. At present, OS is widely investigated because of the interaction between the tumor and bone microenvironment and the effect of such interaction on OS progression and metastasis. The connective tissue growth factor (CTGF), also known as cellular communication network factor 2 (CCN2), is a secreted extracellular matrix-associated protein. CTGF is as active as the regulators of signaling activities of several different pathways and an orchestrator of their cross-talk. Therefore, we conducted experiments to investigate the effects of CTGF on OS tumor progress and the cross-talk with stromal cells in the tumor microenvironment.

1369

Skeletal muscle is the largest motor and metabolic organ of the body, which has a robust capacity for regeneration following injury or disease. Delayed regeneration after skeletal muscle injury reduces muscle contractility and leads to dysfunction of innervation. Therefore, identifying the regulation components in skeletal muscle regeneration and determining their molecular mechanisms are important to discover novel therapeutic markers for muscular diseases. Long non-coding RNA (LncRNA) has been implicated in skeletal muscle regeneration. Recent developed singlecell RNA sequencing (scRNA-seq) provides a higher resolution of cellular differences than bulk RNA-seq. Here, we reanalyzed single-cell transcriptomes data of skeletal muscle regeneration and identified lncRNA maternally expressed gene 3 (lncRNA-MEG3) was highly expressed in muscle satellite cells (MuSCs). Further study showed that lncRNA-MEG3 regulates skeletal muscle regeneration via sponging miR-133a-3p to regulate proline-rich transmembrane protein 2 (PRRT2) expression level. These results suggested that lncRNA-MEG3 might be a potential target for skeletal muscle diseases.

1230

Hepatocellular carcinoma (HCC) is the third leading cause of cancer-related death worldwide. E2 ubiquitin conjugating enzymes (UBE2) are potential therapeutic targets in tumors arising from genomic instability and tumor microenvironment (TME). UBE2S, an important UBE2, has demonstrated strong oncogenic activities in various malignant cancers, including HCC. However, a comprehensive study regarding its role in HCC is still absent, and its association with immunology and drug response of HCC is still unclear. In this study, we conducted a pan-cancer analysis of UBE2S expression and prognosis, carried out enrichment analysis of UBE2S-associated genes, and analyzed association between UBE2S expression and HCC microenvironment, stemness and drug response. Collectively, our results demonstrated that UBE2S expression was significantly increased in multiple types of cancer, including HCC, and harbors prognostic values for HCC. Potential function of UBE2S involves modulation of ubiquitin mediated proteolysis and cell cycle progression. Furthermore, in HCC, UBE2S expression was positively correlated with TME infiltration, microsatellite instability (MSI), DNA methylation, stemness and drug response. These findings highlighted the possible pivotal roles of UBE2S in HCC prognosis, precision immunotherapy and drug response.

1269

Mutations in BRG1-associated factor 45D (BAF45D, also known as double PHD fingers 2 (DPF2)) are linked to Coffin-Siris syndrome (CSS). However, the underlying molecular mechanisms remain ill-defined. Here, we identified that wild-typebutnotCSS-associatedBAF45Dmutants increased the expression of PAX6, a neural stem cell marker, and phosphorylated SMAD3 (p-SMAD3) in spinal cord neural stem cells (NSCs) derived from H9 human embryonic stemcells. BothBAF45DandSMAD3are required for the induction of p-SMAD3 and PAX6 together with STAT3 and SMAD7 by retinoic acid (RA). In the presence of RA, BAF45D knockdown decreased the expression of genes that regulate stem cell pluripotency. Moreover, the same genes in the spinal cord NSCs had more binding sites with BAF45D than those in H9 cells. Furthermore, Gene Ontology (GO) assay results indicated that BAF45D targets more pathways responsible for stem cell maintenance in NSCs. Finally, BAF45D is associated with BRG1, p-SMAD3, and PAX6. Together, these findings suggest a role for BAF45D in modulating PAX6 expression through SMAD signaling, highlighting the importance of BAF45D in neural stem/progenitor cell fate commitment.

1226

Circular RNA (circRNA) is a novel type of noncoding RNA that originates from eukaryotic precursor messenger RNA (pre-mRNA). These circRNAs regulate tumor processes through diverse mechanisms, including serving as a miRNA sponge to modulate expression of miRNA-target genes. Eukaryotic translation initiation factor 3 subunit C (eIF3c) is a core subunit of the IF3 complex, involving in the initiation of the translation process of tumor cells. Currently, little is known about the underlying molecular mechanisms of circEIF3Cinthepathogenesisofintrahepaticcholangiocarcinoma (ICC). A better understanding of the biological functions of circEIF3C in response to tumor cellular growth and how this affects the tumor signaling pathways would provide important clinical value related to the pathogenesis of ICC.

1237

Interleukin-11 (IL-11) is a member of the IL-6 family of cytokines. IL-11 activates its target cells via binding to the IL-11 receptor (IL-11R), which also exists in soluble forms generated by proteolytic cleavage (sIL-11R, required for trans-signaling). Formation of the IL-11/IL-11R complex leads to the recruitment and homodimerization of two molecules of the signal-transducing β-receptor gp130, which then activates intracellular signaling cascades, including the Jak/STAT, PI3K and ERK pathways.

1233

Osteoporosis is a prevalent multifactorial bone disease with a strong genetic contribution. The heritability of traits that contribute to osteoporosis (bone mass, bone mineral density (BMD), bone size, bone loss and fractures) ranges from 50 to 85%, suggesting that a comprehensive understanding of the genetic basis may help identify new therapeutic targets. However, the genetic characteristics remain obscure, and the existing drug targets are associated with various challenges. Numerous studies have demonstrated that high-throughput sequencing data analysis is fruitful for identifying novel targets of human diseases. We therefore integrated GWAS and transcriptome analyses through Multimarker Analysis of GenoMic Annotation (MAGMA) and weighted gene co-expression network analysis (WGCNA) to identify new network modules and potential therapeutic genes for osteoporosis. As an illustration, the flow chart presenting the process of the present study was shown in.

1453

Biallelic loss-of-function variants in ATM (Ataxia Telangiectasia Mutated) cause Ataxia Telangiectasia (AT), a rare disorder associated with cerebellar degeneration and ataxia, cancer predisposition, infertility, growth retardation, etc. ATM is a phosphoinositide 3-kinase-related kinase (PIKK) with a role in DNA repair and maintenance of genome stability. Studying a multisystem genetic disease like AT requires animal models to ascertain its pathogenesis at the level of tissues, organs and the organism. Due to its small size, cheap maintenance, large progeny, rapid development and initial transparency, zebrafish (Danio rerio) is an increasingly popular vertebrate model organism, suitable for genetic modifications and large-scale in vivo therapeutic screens as embryos are chemically permeable to small compounds. Currently, no zebrafish model for AT exists. We generated atm knock-outs through CRIPSR-Cas9 mutagenesis. We show that atm conserved its function as a tumour suppressor gene and is involved in gametogenesis and fertility. Therefore, this mutant is of great value for further studies investigating the role of atm in reproduction and tumorigenesis.

1771

The Coronavirus disease is the contagious disease. Clinical manifests vary from no symptoms, upper and lower respiratory tract infections, and even severe acute respiratory distress syndrome and multiple organs failure. Recently, three of new beta-coronaviruses have emerged to cause serious and widespread illness and death, such as the Severe Acute Respiratory Syndrome Coronaviruses (SARS-CoV), Middle East Respiratory Syndrome Coronaviruses (MERS-CoV), and 2019-nCoV (the causative pathogen of Coronavirus Disease 2019 (COVID-19). The COVID-19 is continuing to spread globally. However, the detailed molecular mechanisms have not yet been fully clarified. Since the similarity of the pathological features among 2019-nCoV, SARS-CoV and MERS-CoV, we could speculate the biological features of current COVID-19 by researching those of MERS-CoV and SARS-CoV. Therefore, we analyzed expression levels of miRNA and mRNA profiles of MERS-CoV, SARS-CoV and 2019-nCoV from five public GEO datasets. As a result, we found 16 miRNAs potentially target a group of hub genes clustering together to tune the immune response. Notably, viral RNAs have the ability to sponge miR-500a-5p and miR-501-5p, indicating viral RNAs sequestering these two miRNAs to trigger genes expression and contribute to pathogenesis. We hope this study will pave the way to better understand 2019-nCoVpathogenesisanddeveloptheinnovative therapeutics.

1261

St. Louis encephalitis virus (SLEV) was first recognized in 1933 and caused a major encephalitis epidemic with 1095 clinical cases and 201 deaths in St. Louis, Missouri. According to the median values from 2014 to 2018, the percentage of neuroinvasion by SLEV was 62% and the fatality rate was 9% in the United States. The clinical symptoms of SLEV infection include neurological abnormalities such as memory loss, disorientation, and meningoencephalitis. Till now, specific treatments including vaccine and drug are unavailable for SLEV infection. It is of particular importance to get prepared for SLEV infection, especially given the currently raging global COVID-19 epidemic.

1227

Nowadays great progress has been made in targeted therapy and comprehensive therapy for a few cancers. But the prognosis of the vast majority of cancer patients is still weakness. It has been proven that inflammation can affect the development and treatment of cancer. Some previous studies have suggested that inflammation-related interleukins (ILs) play a significant role in the development of some cancers. ILs act as a considerable role in immunity, and inflammation affects the development of cancer. Therefore, the expression profile of IL gene may also be one of the vital markers of early pathological changes, occurrence and progression of cancer. Currently, studies on the expression of IL27 in cancer are still extremely limited. Our work comprehensively and systematically studied the IL27 gene expression in the generic cancer when compared with the existing IL27 gene related research experiment. We screened out cancer species linked with poor prognosis, and explored the correlation between IL27 gene expression and immune cell infiltration and immune microenvironment in these cancers further, providing an increasing number of the direct basis for subsequent studies. Our findings showed that IL27 can be used to assess prognosis and immune infiltration in patients with colon adenocarcinoma (COAD), glioblastoma multiforme (GBM), head and neck squamous cell carcinoma (HNSC), skin cutaneous melanoma (SKCM) and testicular germ cell tumors (TGCT) as a biomarker.

1257

1250

The key pathogenesis of chronic myeloid leukemia (CML) is the formation of BCR-ABL fusion gene, encoding a 210 kDa Bcr-Abl tyrosine kinase, which is crucial for the occurrence and development of CML. Imatinib (IM) is the first targeted anticancer drug approved by FDA for the treatment of CML; however, some patients, especially those in accelerated phase and blastic phase, develop primary or secondary drug resistance to IM. Particularly, the most challenging resistance is caused by T315I mutation of Bcr-Abl, which represents approximately 15%-20% of all acquired mutations and renders cell resistant to a variety of tyrosine kinase inhibitors. Thus, there is an urgent need to develop novel strategiestoovercomeBcr-AblT315I-meidatedIM resistance.

1306

Atherosclerosis, the underlying pathophysiological basis of cardiovascular disease, has been recognized as a lipid-driven chronic inflammatory disease. Sterol carrier protein 2 (SCP-2) is a 13-kDa non-specific lipid-transfer protein expressed by various tissues and cells, such as liver, heart, vascular smooth muscle cells (VSMCs), and macrophages. SCP-2 has an extensive role in cardiovascular and metabolic diseases. Recently, SCP-2 was reported to promote the development of atherosclerosis by regulating lipid metabolism and peroxidation, endocannabinoid metabolism, vascular inflammation, and fatty acid metabolism. In this review, we summarized the recent advances regarding the role of SCP-2 in the pathogenesis of atherosclerosis and tried to provide a rationale for future investigation and a better understanding of the biological functions of SCP-2 in atherosclerotic cardiovascular disease.

1411

Genetic compensation is a remarkable biological concept to explain the genetic robustness in an organism to maintain its fitness and viability if there is a disruption occurred in the genetic variation by mutation. However, the underlying mechanism in genetic compensation remain unsolvable. The initial concept of genetic compensation has been studied in model organisms when there was a discrepancy between knockout-mediated and knockdown-mediated phenotypes. In the zebrafish model, several studies have reported that zebrafish mutants did not exhibit severe phenotype as shown in zebrafish morphants for the same genes. This phenomenon in zebrafish mutants but not morphants is due to the response of genetic compensation. In 2019, two amazing works partially uncovered genetic compensation could be triggered by the upregulation of compensating genes through regulating NMD and/or PTC-bearing mRNA in collaboration with epigenetic machinery in mutant zebrafish. In this review, we would like to update the recent advances and future perspectives of genetic compensation studies, which including the hypothesis of time-dependent involvement and addressing the discrepancy between knockout-mediated and knockdown-mediated to study gene function in the zebrafish model. At last, the study of genetic compensation could be a potential therapeutic strategy to treat human genetic disorder related diseases.

1946

The liver injury leads to an inflammatory response, which causes the activation of hepatic stellate cells (HSCs) that further secrete ECM proteins and play an important role in liver fibrosis. Moreover, the inflammatory response is a driving force for fibrogenesis, which is triggered by many types of injuries. Exaggerated inflammatory immune responses are mediated by cytoplasmic protein complexes known as inflammasomes, which are involved in many chronic liver diseases. Inflammasomes are pattern recognition receptors (PRRs) that can sense any mi-crobial motifs known as pathogen-associated molecular patterns (PAMPs), and host-or environmental-derived stress signals known as damage-associated molecular patterns (DAMPs). The inflammasomes cause caspase-mediated proteolytic cleavage of pro-IL-1β and pro-IL-18 into active IL-1β and IL-18. In this review, we provide a comprehensive summary of the important roles of NLRP3 inflammasome in the pathogenesis of liver fibrosis with an emphasis on several direct and indirect pathways responsible for the NLRP3 inflammasome-mediated HSCs activation and fibrogenesis. In addition, we discuss the general pharmacological and genetics strategies for the inhibition of NLRP3 inflammasome activation and its downstream signaling with examples of emerging pharmacotherapeutics, targeting the NLRP3 inflammasome signaling as well as a possible way to develop effective and safer NLRP3 inflammasome inhibitors.

1390

N⁶-methyladenosine (m⁶A) modification is found the most prevalent and abundant post-transcriptional mRNA modification in eukaryotic cells. It regulates almost all stages of RNA life cycle including splicing, translocation, stability, decay and translation. As a dynamic and reversible process, m⁶A modification is catalyzed by the RNA methyltransferases ('writers'), removed by the demethylases ('erasers'), and interacts with m⁶A-binding proteins ('readers'). Recent studies have revealed that these m⁶A modification regulators are frequently expressed aberrantly in various types of cancer, and involved in cell proliferation, differentiation, metabolism, particularly, in tumorigenesis and tumor progression through diverse mechanisms. In this review, the m⁶A modification process and its regulatory functions in lung cancer are summarized. Furthermore, the research progress in the inhibitor development of m⁶A modification, and the potential of targeting m⁶A modifying proteins for clinical application are discussed.

1291

There are many complex eye diseases which are the leading causes of blindness, however, the pathogenesis of the complex eye diseases is not fully understood, especially the underlying molecular mechanisms of N6-methyladenosine (m6A) RNA methylation in the eye diseases have not been extensive clarified. Our review summarizes the latest advances in the studies of m6A modification in the pathogenesis of the complex eye diseases, including cornea disease, cataract, diabetic retinopathy, age-related macular degeneration, proliferative vitreoretinopathy, Graves’disease, uveal melanoma, retinoblastoma, and traumatic optic neuropathy. We further discuss the possibility of developing m6A modification signatures as biomarkers for the diagnosis of the eye diseases, as well as potential therapeutic approaches.

1315

Severe bronchopulmonary dysplasia (BPD) is a chronic lung disorder that primarily affects premature babies with extremely low birth weight and involves in multiple organ system; no effective pharmacotherapy for this disease exists, and mortality remains high. Based on the evidence from previous preclinical studies and phase I clinical trials, this study aims to test the safety of intravenous application of a single dose of human umbilical cord-derived mesenchymal stem cells (hUC-MSCs) in patients with severe BPD. The Mesenchymal Stem cells for Bronchopulmonary Dysplasia Treatment (MSBDT) trial is a single center, open-label, doseescalation phase I clinical trial. Severe BPD patients were enrolled in Children Hospital of Chongqing Medical University, Chongqing, China. The first six patients were treated with low-dose hUC-MSCs (1 × 10⁶ cells/kg) and the next seven patients were treated with high-dose hUC-MSCs (5 × 10⁶ cells/kg). This study is registered with ClinicalTrials. gov, number NCT03558334. No prespecified infusion-associated adverse events, immediate complication, respiratory or cardiovascular compromise were observed during infusion and 24 h after infusion. No significant changes in safety laboratory values were observed. One death event occurred in the low-dose group on study day 10, and one death event occurred in the high-dose group on study day 24, while, after review in detail, the two cases are not believed to be infusion-associated events. In conclusion, intravenous application of a single dose of hUC-MSCs was tolerated in thirteen patients with severe BPD.

1293

Accumulating evidence indicates that RNA methylation at N⁶-methyladenosine (m6A) plays an important regulatory role in gene expression and aberrant mRNA m6A modif ication is often associated with a variety of cancers. However, little is known whether and how m6A-modification impacts long non-coding RNA (lncRNA) and lncRNA-mediated tumorigenesis, particularly in pancreatic ductal adenocarcinoma (PDAC). In the present study, we report that a previously uncharacterized lncRNA, LINC00901, promotes pancreatic cancer cell growth and invasion and moreover, LINC00901 is subject to m6A modification which regulates its expression. In this regard, YTHDF1 serves as a reader for the m6A modified LINC00901 and downregulates the LINC00901 level. Notably, two conserved m6A sites in LINC00901 are critical to the recognition of LINC00901 by YTHDF1. Finally, RNA sequencing (RNA-seq) and gene function analysis revealed that LINC00901 positively regulates MYC through upregulation of IGF2BP2, a known RNA binding protein that can enhance MYC mRNA stability. Together, our results suggest that there is a LINC00901-IGF2BP2-MYC axis through which LINC00901 promotes PDAC progression in an m6A dependent manner.

1361

Our previous studies found that Zinc-finger protein 382 (ZNF382) played as a tumor suppressor gene in esophageal and gastric cancers, and a positive correlation between the high expression of ZNF382 and better outcome in breast cancer patients. However, the biological roles and mechanisms of ZNF382 in breast cancer remains unclear. We detected ZNF382 expression by reverse-transcription PCR (RT-PCR) and real-time quantitative PCR (qRT-PCR) in breast cancer cells and tissues, and explored the impacts and mechanisms of ectopic ZNF382 expression in breast cancer cells in vitro and in vivo, respectively. Our results revealed that ZNF382 was significantly down-regulated in breast cancer tissues compared with adjacent non-cancer tissues. Restoration of ZNF382 expression in silenced breast cancer cells not only inhibited tumor cell colony formation, viability, migration and invasion, and epithelial-mesenchymal-transition (EMT), but also induced apoptosis and G0/G1 arrest. In conclusion, ZNF382 could induce G0/G1 cell cycle arrest through inhibiting CDC25A signaling, and, inhibit cell migration, invasion and EMT by antagonizing ZEB1 signaling in breast cancer cells.

1291

Atherosclerosis is a chronic inflammatory disease, occurring preferentially in bifurcation, branching, and bending of blood vessels exposed to disturbed flow. Disturbed flow in atheroprone areas activates elevated proteases, degrading elastin lamellae and collagenous matrix, resulting in endothelial dysfunction and vascular remodeling. As a mediator for extracellular matrix protein degradation, cathepsin K (CTSK) was directly regulated by hemodynamics and contributed to atherosclerosis. The mechanism of CTSK responding to disturbed flow and contributing to disturbed flow-induced atherosclerosis is unclear. In this study, the partial carotid ligation model of mice and in vitro disturbed shear stress model were constructed to explore the contribution and potential mechanism of CTSK in atherosclerosis. Our results indicated that CTSK elevated in the disturbed flow area in vivo and in vitro along with endothelial inflammation and atherogenesis. Additionally, the expression of integrin avb3 was upregulated in these atheroprone areas. We found that inhibition of the integrin avb3-cytoskeleton pathway could significantly block the activation of NF-κB and the expression of CTSK. Collectively, our findings unraveled that disturbed flow induces increased CTSK expression, and contributes to endothelial inflammation and vascular remodeling, leading to atherogenesis eventually. This study is helpful to provide new enlightenment for the therapy of atherosclerosis.

1313

With the development of tyrosine kinase inhibitor (TKI) resistance, finding the novel effective chemotherapeutic agent is of seminal importance for chronic myelogenous leukemia (CML) treatment. This study aims to find the effective anti-leukemic candidates and investigate the possible underlying mechanism. We synthesized the novel coumarin derivatives and evaluated their anti-leukemic activity. Cell viability assay revealed that compound DBH2 exhibited the potent inhibitory activity on the proliferation of CML K562 cells and TKI resistant K562 cells. Morphological observation and flow cytometry confirmed that DBH2 could selectively induce cell apoptosis and cell cycle arrest at G2/M phase of the K562 cells, which was further confirmed on the bone marrow cells from CML transgenic model mice and CD34⁺ bone marrow leukemic cells from CML patients. Treatments of DBH2 in combination with imatinib could prolong the survival rate of SCL-tTA-BCR/ABL transgenic model mice significantly. Quantitative RT-PCR revealed that DBH2 inhibited the expression of STAT3 and STAT5 in K562 cells, and caspase-3 knockout alleviated the DBH2 induced apoptosis. Furthermore, DBH2 could induce the expression of PARP1 and ROCK1 in K562 cells, which may play the important role in caspase-dependent apoptosis. Our results concluded that coumarin derivative DBH2 serves as a promising candidate for the CML treatment, especially in the combination with imatinib for the TKI resistant CML, and STAT/caspase-3 pathway was involved in the molecular mechanism of anti-leukemic activity of DBH2.

1285

Inactivated COVID-19 vaccines have been widely used to vaccinate the Chinese population. However, limited literature exists to explore the effect of obesity on the humoral and cellular immune response to these vaccines. In this study, 132 high BMI (Body mass index) (obesity and overweight, BMI ≥ 24 kg/m²) and 82 normal BMI (BMI<24 kg/m²) participants were enrolled. Adverse events (AEs), Spike receptor-binding domain IgG antibody (anti-RBD-IgG), neutralizing antibodies (NAbs), and specific B-cell and T-cell responses were evaluated 21-105 days after full-course inactivated COVID-19 vaccination. The overall incidence of AEs was similar in individuals with and without obesity/overweight. No serious vaccine-related AEs occurred. Individuals with obesity/overweight had a reduced seropositivity rate of NAbs compared to those with normal BMI. Anti-RBD-IgG and NAbs titers in the high BMI group were significantly lower than those in the normal BMI group. The frequencies of RBD-specific memory B cells (MBCs) and the numbers of spike-specific TNF-α+ spot-forming cells (SFCs) in individuals with obesity/overweight were reduced compared with those noted in individuals without obesity/overweight. A similar trend of weakened humoral responses was also observed in individuals with central obesity. Our study results suggested that inactivated COVID-19 vaccines were safe and well tolerated but induced poor humoral and cellular immune responses in Chinese individuals with obesity/overweight.

1949