第10卷，第6期

Triple-negative breast cancer (TNBC) has a poor prognosis because of its aggressive characteristics and lack of targeted therapies. Recently, it has been reported that TNBC is sensitive to ferroptosis, an iron-dependent type of programmed cell death, making it a potential target for the treatment of TNBC. Breast cancer stem cells (BCSCs), a small population of cancer cells that possess the infinite proliferative potential and tumor-initiating properties, play important roles in drug resistance. The responses of BCSCs to ferroptosis-inducing agents are controversial and remain elusive.

1277

Breast cancer has been one of the biggest killers of women due to its susceptibility and high metastasis. Pathological observations show that malignant cancer cells frequently invade the surrounding normal tissue in collective rather than individual cell migration. For individual cell migration, it has been found that the Rho/ROCK signaling is upregulated and correlates with disease progression. Meanwhile, Rho activates myosin-II and the actomyosin-mediated contraction creates tension within the cells. However, the roles of the activation of the Rho/ROCK signal pathway in collective cell migration and the precise mechanisms by which myosin-II fine-tunes the contractility of the cells to allow for the reorganization of the cytoskeleton that drives collective cell migration, remain unclear. This study investigated whether the high cellular contractility could activate the inside-out signal trans-ductions and how did the intracellular force contribute to the collective cell migration.

1259

Despite the spread of effective vaccination strategies, cervical cancer remains the second leading cause of cancer death among women aged 20 to 39. Clinical diagnosis of cervical lesions relies on the P16INK4a (P16) marker, but its sensitivity to low-grade cervical intraepithelial neoplasia (CIN) is limited. Exploring more sensitive and specific molecular markers is still a challenge in cervical lesion screening. In this study, we found that HMGB3 could effectively label pathological cells in different cervical lesions, especially for early CIN. Therefore, HMGB3 has the potential to be used as a novel marker for the early screening of cervical lesions.

1333

Diabetes mellitus (DM) is one of the most common diseases in the elderly, and among DM patients, more than 90% have type 2 DM (T2DM). The etiology of T2DM is complex and associated with risk factors such as age, genetic disposition, and diet. The genetic factors underlying T2DM are currently the focus of intense research, and it has been established that the susceptibility genes associated with T2DM vary among different populations. A recent longitudinal exome-wide association study in the Japanese population has identified GGA3 as a susceptible locus for T2DM. GGA3 is one of the Golgi-localized g-ear-containing ARF binding proteins (GGAs) that function widely in the transport of Golgi-derived vesicles and in endocytic trafficking pathways. However, although it is widely acknowledged that the control of glucose homeostasis via endocytosis and glucose metabolism plays a vital role in T2DM, the associations between intracellular trafficking and diabetes pathogenesis have not been sufficiently established. In this study, we used a Gga3 gene knockout mouse strain to investigate the potential involvement of GGA3 in controlling blood glucose. The study was approved by the ethics committee of Jining Medical University (No. 2019-JS-005). Our results indicate that the GGA3 gene might play a protective role against diabetes.

1387

Malaria caused by the Plasmodium falciparum parasite is responsible for more than 240 million cases per year and killed 627, 000 people in 2020, mostly African children. The malaria parasite is transmitted by mosquitos belonging to the genus Anopheles. After an asymptomatic liver stage, the parasite is released into the bloodstream to invade red blood cells (RBCs) and replicate asexually. This erythrocytic phase is associated with a variety of clinical manifestations, including mild and severe malaria. Cerebral malaria (CM) is one of the most severe forms, characterized by the sequestration of parasitized RBCs in the small capillaries of the brain and the local development of cytokine-mediated inflammation. Genetic variants in genes encoding proteins involved in red blood cell physiology are protective factors against severe malaria, as clearly demonstrated for the sickle cell variant of hemoglobin (HbS). Rare gain-of-function mutations in Piezo1, a mechanosensitive calcium channel, are involved in hereditary xerocytosis, a disease characterized by red cell dehydration and mild hemolysis. Interestingly, RBC dehydration is associated with reduced Plasmodium infection in vitrosuggesting that PIEZO1 polymorphisms may protect against malaria. Recently, a gain-of-function PIEZO1 E756del variant (rs59446030) under positive selection in Africa, was associated with protection against severe malaria in children in Gabon. However, a large caseecontrol study in Ghana failed to replicate this association. In this study, we sought to further characterize the role of this PIEZO1 polymorphism in malaria by confirming its association in a Senegalese population and identifying the genetic interaction with polymorphisms in the ATP2B4 gene that encodes PMCA4, the major RBC calcium pump. We prove that activation of Piezo1 modulates intracellular calcium concentration and inhibits Plasmodium falciparum multiplication in RBCs. Furthermore, we demonstrated the additive effect of Piezo1 activation and PMCA4 blockade on the inhibition of parasite multiplication in erythrocytes.

1303

Acute myeloid leukemia (AML) and acute lymphoblastic leukemia (ALL) are distinct subtypes of acute leukemia with respect to clinical and genetic features. Recently, ALLs were reported to share similar genes like IDH1 mutations to AMLs. Specifically, mutated IDH1 (5%-15%) frequently occurs in AML and scarcely (1.9%) in ALL. Notably, IDH1 along with SRSF2 mutation often exists in secondary AML patients and confers a poor prognosis. Recently, venetoclax (VEN) has been demonstrated extremely effective in combination with a hypomethylating agent or chemotherapy for IDH1 mutated AML patients. However, whether VEN combining chemotherapy is effective among ALL patients with similar genetic features to AML remains unclear. Here, we successfully rescued a relapsed ALL case with IDH1 mutation by VEN plus methotrexate and pegaspargase based on their antimetabolic pathways.

1458

In recent years, increasing attention has been given to public health. Cancer and obesity are hot topics, in which glucose and lipid metabolism play an important role. Glycogen synthase GYS2 has been found to regulate glycogen metabolism in hepatocellular carcinoma (HCC) and is a potential prognostic factor and therapeutic target. Excessive bile acids induce intestinal injury and inflammatory bowel disease, and targeting bile acid receptors (FXRs) can regulate inflammation and repair damaged intestinal epithelial barriers. However, the research model of metabolic pathways focuses on the key enzymes, while ignoring the overall relevance. Here, we targeted the glycogen anabolism pathway and screened the upstream factor, bile acid transporter SLC10A1.SLC10A1 participates in the enterohepatic circulation of bile acids and regulates the transcriptional activity of the bile acid receptor FXR. Besides, we found that diazinon could bind NTCP and inhibit glycogen synthesis by mimicking the disordered bile acid enterohepatic circulation. Collectively, enterohepatic circulation of bile acids coordinates lipid metabolism and glucose metabolism and draws the metabolic crosstalk network, which provides a new perspective for the clinical treatment of HCC.

1335

1425

Tumor-associated lymphangiogenesis is thought to be an important metastatic step in intrahepatic cholangiocarcinoma (ICC), while the origin of tumor-associated lymphatic vessels within tumors is little known. Cancerassociated fibroblasts (CAFs) enhance tumor progression through many aspects. CAFs have been discovered exhibiting cellular plasticity of phenotypic transition - such as epithelialemesenchymal transition (EMT) - which promotes increased migratory and invasive capabilities of cells. However, mesenchymal-to-endothelial (MEndT) transition is yet to be identified in tumor entities. Podoplanin (PDPN), a 38-44 kDa transmembrane glycoprotein, is conventionally expressed in lymphatic endothelial cells. Recently, PDPN has also been discovered as a marker of CAFs, which is associated with cancer cell migration, invasion, and clinical prognosis. In several types of tumors such as breast cancer and lung adenocarcinoma, it was demonstrated that PDPN-expressing CAFs are correlated with tumor progression and poor prognosis. Based on abundant CAFs in ICC and PDPN expression in both CAFs and lymphatic endothelial cells, our study aims to investigate whether PDPN-positive CAFs could transform into lymphatic endothelial cells contributing to lymphangiogenesis in ICC.

1309

The second leading cause of death worldwide is cancer. Cancer is a general term that refers to a highly diverse and complex set of over 200 diseases where abnormal cells grow uncontrollably and avoid death. These diseases can start almost anywhere within the body and may develop the ability to invade nearby tissues and spread. Every cancer is the byproduct of a unique combination of various genetic and environmental factors. Even within the same individual, different cells may possess distinct cancerous mutations which can mutate further over time. Despite this inherent heterogeneity, the most common treatments for cancer are surgery, chemotherapy, and radiation.

1518

Severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2), which causes coronavirus disease 2019 (COVID-19) and initially infects epithelial cells of the upper respiratory tract, has recently caused a global health emergency and is still a potential threat to humans. Multiple studies have suggested alterations in the immune system, including excessive neutrophil activation and lymphopenia, and excessive inflammatory responses as the key mechanisms forsevere symptoms. However, which cell types contribute to excessive inflammation remains to be explored. Further analysis of human blood immune cells provides insights into the coordinated response to SARS-CoV-2 infections and is a significant task for restraining the virus and controlling disease progression. In this study, we reanalyzed the single-cell sequencing (scRNA-seq) dataset GSE157789 uploaded by Sinha S et al in the GEO database, and the results were validated by analyzing the RNA-seq dataset GSE171110. The results indicated increased neutrophils and plasma cells but reduced CD8 T cells. More importantly, we identified a unique population of fibroblasts enriched in severe COVID-19. The fibroblast subcluster showed potential neutrophil-like function and might be another significant cell population participating in uncontrolled inflammation in COVID-19.

1235

Emerging evidence indicates that metabolism reprogramming plays an important role in cancer progression. RNAbinding protein nucleolin (NCL) was reported to function as an important oncogenic factor in multiple cancer types. However, the role and mechanism of NCL in cancer metabolism are unknown. In this study, we found that NCL directly interacted with hnRNPA1 and promoted CRC cell proliferation by enhancing aerobic glycolysis. Mechanistically, NCL bound PKM pre-mRNA and increased hnRNPA1-mediated PKM alternative splicing, resulting in increased PKM2 expression and tumor growth. A seed-mediated growth approach was used to synthesize gold nanostars (GNS), which were further modified with aptamer AS1411 (an NCL ligand), GE11 (an EGFR ligand), and nuclear localization signal to obtain functionalized nanoparticles (GNS-AS1411-GE11). GNS-AS1411-GE11 efficiently enter the nucleus of CRC cells and blocked the glycolysis-promoting effects of NCL, inhibiting the growth of CRC xenograft. Targeting NCL is a promising strategy for treating CRC.

1263

Around 10%-30% of non-small cell lung cancer (NSCLC) patients harbored epidermal growth factor receptor (EGFR) mutations, with L858R and exon-19 deletions (19-Del) accounting for 90% of cases. EGFR tyrosine kinase inhibitors (TKIs) showed significant efficacy against common EGFR mutations. However, the therapeutic relevance of uncommon EGFR mutations remained insufficiently investigated. EGFR fusions are extremely rare (0.05%-0.13%) in NSCLC, and Konduri group reported only 5 EGFR fusions from 10, 097 patients. Additional EGFR fusions were reported in NSCLC, all of which were oncogenic drivers and sensitive to EGFR TKIs. Herein, we reported an NSCLC patient with leptomeningeal metastasis (LM) who acquired a novel EGFR-SEPT14 fusion during TKI resistance and showed promising responses to certain EGFR TKIs and intrathecal pemetrexed (IP).

1237

The two most common inherited forms of colorectal cancer (CRC) are familial adenomatous polyposis and hereditary nonpolyposis colorectal cancer. They are caused by germline mutations in the APC and mismatch repair genes, respectively. The simultaneous inheritance of both an APC gene mutation and a mismatch repair gene mutation is very rare. In this study, we reported a coinheritance of mutations in the APC and MLH1 genes in a 20-year-old patient with CRC and multiple polyposes. We described for the first time the somatic mutational profile of a colorectal tumor harboring concurrent germline mutations in the APC and MLH1 genes.

1284

Tauopathies, such as Alzheimer's disease (AD), are neurodegenerative diseases characterized by the deposition of neurofibrillary tangles comprising hyperphosphorylated tau protein in the human brain. Given that abnormal epigenetic alterations in heterochromatin configuration have been documented in AD patients and transgenic animal models of AD, we investigated the roles of novel heterochromatin-associated interactors in tauopathies. Using transgenic flies via UAS-Gal4 binary system, we found that knockdown of Hipp1 (HP1a and insulator partner protein-1) ameliorates tau^R406W (referred to as "tau" hereafter for simplicity) -induced locomotion defects, reduced lifespan, and degeneration of brain tissues. Intriguingly, Hipp1 knockdown restored tau-driven aberrant expression of putative insulator targets and aberrant insulator-mediated epigenetic alterations. HIPP1 may have a role as an insulator-binding partner regarding being implicated in tauinduced neurodegeneration. Moreover, Hipp1 knockdown in flies overexpressing tau restored the aberrant expression of AD susceptibility genes. These observations provide new insights into the roles of insulator proteins in tauopathies.

1889

Rheumatoid arthritis (RA) is a chronic inflammatory disease characterized by cartilage and bone damage with the presence of pannus formation which causes uncontrollable proliferation and invasion of fibroblast-like synoviocytes (FLS). Since rheumatoid arthritis is a chronic disorder, the patients normally need to undergo prolonged antirheumatic treatment with the use of disease-modifying antirheumatic drugs (DMARDs), steroids, and nonsteroidal anti-inflammatory drugs (NSAIDs). The efficacy of such long-term pharmaceutical intervention can be significantly affected by the development of drug resistance. The pathological relationship between rheumatoid arthritis and cancer hinted that some chemotherapeutic drugs, such as methotrexate (MTX), could be used for RA treatment. This idea was reinforced by the analysis of mutations in p53 tumor suppressor gene. Around 50% of p53 somatic mutations observed from various cancers are also identified in the synovium of RA patients. Of note, the hyperplastic synovium in RA with overexpressed p53 mutants contributed to the destruction of joints in patients with erosive RA. Amongst the different cellular components in the synovium of RA patients, p53 mutants are overexpressed in RA synovial fibroblasts (RASFs), which is the pathological phenotype of the FLS..

1254

Tumor microenvironment (TME) is the environment around the tumor cells, consisting of extracellular matrix, nonmalignant stromal cells, and various types of immune cells (like macrophages, B and T lymphocytes). TME has been recurrently reported to play important roles in the progression of cancers. The mechanism study of TME may offer opportunities for targeted/immune therapies. However, how TME components impact the prognosis of cancer patients remains to be determined. In this study, we constructed a web server named OStme (Online consensus Survival analysis web server for Tumor MicroEnvironment components of pan-cancers) to analyze the association of cancer prognosis and various types of tumor-infiltrating cells (TICs), and to investigate the fraction variations of TICs across different stages/grades or variations along with metastasis/recurrence or variations regarding therapies. OStme is freely available at http://bioinfo.henu.edu.cn/Immune/Immune.html. OStme provides 12 types of survival terms (such as OS, DSS, and RFS), offers four analysis modules for cancer patient survival analysis, differential analysis per clinical characteristics (such as different stage, tumor versus normal), comprehensive TME fraction estimation, and the correlation between gene expression and TIC abundance of pan-cancers. We believe OStme will serve as a valuable resource and tool to systematically evaluate the impact of TICs on the prognosis, metastasis, recurrence, progression, and therapy of pan-cancers.

1260

Histone methyltransferase enzyme SET7 and mixed-lineage leukemia protein (MLL) complex are crucial co-activators of androgen receptor (AR) and have recently emerged as potential therapeutic targets for advanced castration-resistant prostate cancer (CRPC). In this study, we described the identification of a rhodium-based hybrid complex (SM_1) as a potent blocker of AR activity via simultaneously inhibiting SET7 and MLL complex activity, which makes it a potential lead scaffold for CPRC drug development.

1257

Krabbe disease or globoid cell leukodystrophy (GLD; MIM#245200) is a rare and fatal lysosomal storage disease with an autosomal recessive mode of inheritance that results from the deficiency of galactocerebrosidase (GALC; E.C. 3.2.1.46), a lysosomal enzyme encoded by the GALC gene. GALC breaks down galactosylceramide, a cerebroside located mainly in the myelin sheath. Defects in GALC cause the accumulation of a cytotoxic metabolite, galactosylsphingosine or psychosine, which can be toxic to oligodendrocytes and Schwann cells. The failure to digest galactosylceramide triggers the formation of multi-nucleated globoid cells, causing severe demyelination, axonopathy, and neuronal death. The reported frequency of Krabbe disease is 1 in 100,000 live births with symptoms including irritability, loss of motor ability, spasticity, ataxia, visual dysfunction, seizures, and cognitive impairment.

1264

S-phase kinase-associated protein 2 (Skp2) is a well-characterized oncoprotein localized mainly in the nucleus and cytoplasm. It is an integral component of SCF^Skp2 E3 ubiquitin ligase complex which confers substrate selectivity to the ligase by specifically targeting a distinct set of proteins destined for proteasomal degradation such as p21, p27, cyclin E, and c-Myc. Skp2 is crucial in a multitude of cellular processes including cell cycle, cell proliferation, apoptosis, differentiation, and survival. However, despite its immense and well-established role in ubiquitin-proteasome system-mediated protein turnover, much is unknown about the function of Skp2 independent of the ubiquitination pathway. Previously, Skp2 has been reported to regulate RhoA gene transcription and the p300 signaling pathway in an E3 ligase-independent manner. Moreover, Skp2 also acts as a cofactor for c-Myc-regulated gene expression.

1296

Colorectal cancer (CRC) is known to harbor considerable heterogeneity. Consequently, it could be hypothesized that similar-appearing tumors might exhibit substantial genetic differences while diverse-appearing tumors may have a similar genetic landscape. Due to these differences at the molecular level, they behave or respond differently to therapies as well. CRC progression is a multistep process and involves the accumulation of substantial genetic and epigenetic events in a stage-dependent manner. Alterations in Wnt, DNA repair, RAS-RAF-MAPK, and PIK3CA-AKT pathways have been well-established to play a role in the etiology of CRC. In the era of personalized medicine, it becomes essential to identify the molecular subtype of CRC so that the predictive and prognostic potential of CRC could be established. Molecular subtyping has its importance and limitations in the clinical management of CRC and is very complex to comprehend. Thus, there is still a need to create robust and reliable clustering methods that could precisely identify unique molecular signatures and help in the prediction of the clinical response of the patients who share certain clinical as well as molecular characteristics. Frequent mutations have been reported in RAS, BRAF, NRAS, and PIK3CA genes that are major regulators of the above pathways and significantly promote tumor initiation and progression in CRC. Similarly, the clinical significance of epigenetically deregulated MLH1, RASSF1, DAPK1, IFG2, SLITRK5, and IGFBP3 genes is also well-established and documented in CRC. In the present investigation, we have comprehensively analyzed the mutation status of KRAS, BRAF, NRAS, and PIK3CA, and methylation status of MLH1, RASSF1, SLITRK5, DAPK1, IGFBP3, and IGF2 genes in 70 CRC tumor samples and 40 matched normal, which have a significant role in CRC initiation and progression. Further, the patients were stratified according to their shared molecular heterogeneity and the prognostic value of each heterogeneous cluster was evaluated.

1229

Lung squamous cell carcinoma (LSCC) is the second most common histological type of non-small cell lung cancer (NSCLC) after lung adenocarcinoma, accounting for 30% of all NSCLC cases. Cytotoxic chemotherapy and radiotherapy are commonly used as adjuvant therapy following surgery for LSCC. Advanced and metastatic LSCC patients typically receive systematic therapy that consists of a platinumbased doublet. Rare mutations of two main oncogenic drivers (EGFR and KRAS) in LSCC hindered the development of molecular targeted therapies. Cancer immunotherapy based on anti-PD-1 (programmed cell death 1) /PD-L1/CTLA-4 (cytotoxicT-lymphocyte-associatedprotein4) therapies represents promising therapeutics for NSCLC patients, while obtained remission rate was unsatisfactory partly due to the heterogeneity of the tumor immune microenvironment (TIME). Therefore, comprehensive characterization of TIME in LSCC is vital to predict the responsiveness to immunotherapy and develop new effective therapies for LSCC.

1306

Endometrial cancer (EC) is the second most prevalent female reproductive system tumor after cervical cancer and its incidence is rising globally. Recently, N6-methyladenosine (m⁶A) has emerged as an important regulator of a variety of physiological and pathological processes, especially in promoting EC progression. However, the potential roles of m⁶A modification in EC and its immune landscape remain unknown. Here, the correlation between m⁶A modification patterns and tumor microenvironment (TME) cell-infiltrating characteristics was comprehensively evaluated based on 21 m⁶A regulators of 1301 endometrial cancer samples. We identified four m6Aclasses characterized by distinct TME cell infiltration. We used principal component analysis algorithms to construct m6Ascore to quantify m⁶A modification patterns of endometrial cancer individuals. Our results showed that lower m6Ascore may be closely related to immune-inflamed phenotype and displayed immune-activated characteristics and better prognosis, while higher m6Ascore may be associated with immune-desert and immune-excluded phenotype and showed interstitial activation-related characteristics and poor prognosis. These new findings reveal that m⁶A modification played a nonnegligible role in the formation of TME diversity and complexity and support the possibility of targeting m⁶A modification patterns for rescuing EC, which were unreported in EC patients.

1353

Piwi-interacting RNAs (piRNAs of 26-32 nt in length) regulate gene expression, epigenetics, and transcriptional and post-transcriptional processes. In humans, piRNAs are expressed in germline and somatic tissues, and generally have a uracil at the 5′-end (position +1), an adenine at the +10 position, and are 2′-O methylated at the 3′-end. Onice piRNAs form the RNA-induced silencing complex (RISC) with PIWI proteins, they serve as guides to find their complementary sequences of the target mRNA, thus initiating their degradation. The Wuhan patient's genome is approximately 265 nucleotides (nts) in its 3′-UTR; piRNAs-like sequences have been reported in this region. Our objective was to search, through bioinformatics, for mRNA sequences that were homologous to the reverse complementary of the previously reported piRNA-like sequences from the 3′-UTR of SARS-CoV-2, and identify the possible interaction between them.

1330

Autophagy is an evolutionarily conserved process involved in the degradation of long-lived proteins and excessive or dysfunctional organelles. As a pivotal cellular response, autophagy has been extensively studied and is known to be involved in various diseases. Ferroptosis is a recently discovered form of regulated cell death characterized by iron overload, leading to the accumulation of lethal levels of lipid hydroperoxides. Recently, an increasing number of studies have revealed a link between autophagy and ferroptosis. Myocardial ischemia/reperfusion injury (MIRI) is an urgent dilemma after myocardial infarction recanalization, which is regulated by several cell death pathways, including autophagy and ferroptosis. However, the potential relationship between autophagy and ferroptosis in MIRI remains unexplored. In this study, we briefly review the mechanisms of autophagy and ferroptosis, including their roles in MIRI. Moreover, we provide an overview of the potential crosstalk in MIRI. Clarifying the relationship between different cell death pathways may provide new ideas for the treatment of MIRI in the future.

1413

Virus-related cancer is cancer where viral infection leads to the malignant transformation of the host's infected cells. Seven viruses (e.g., human papillomavirus (HPV), Epstein eBarr virus (EBV), Kaposi's sarcoma herpesvirus (KSHV), Hepatitis B virus (HBV), Hepatitis C virus (HCV), Human T-lymphotropic virus (HTLV), and Merkel cell polyomavirus (MCV)) that infect humans have been identified as an oncogene and have been associated with several human malignancies. Recently, growing attention has been attracted to exploring the pathogenesis of virus-related cancers. One of the most mysterious molecules involved in carcinogenesis and progression of virus-related cancers is circular RNAs (circRNA). These emerging non-coding RNAs (ncRNAs), due to the absence of 5′and 3′ends, have high stability than linear RNAs and are found in some species across the eukaryotic organisms. Compelling evidence has revealed that viruses also encode a repertoire of circRNAs, as well as dysregulation of these viral circRNAs play a critical role in the pathogenesis and progression of different types of virusrelated cancers. Therefore, understanding the exact role and function of the virally encoded circRNAs with virus-associated cancers will open a new road for increasing our knowledge about the RNA world. Hence, in this review, we will focus on emerging roles of virus-encoded circRNAs in multiple cancers, including cervical cancer, gastric cancer, Merkel cell carcinoma, nasopharyngeal carcinoma, Kaposi cancer, and liver cancer.

1340

The bromodomain and extra-terminal (BET) proteins act as "readers" for lysine acetylation and facilitate the recruitment of transcriptional elongation complexes. BET protein is associated with transcriptional elongation of genes such as c-MYC and BCL-2, and is involved in the regulation of cell cycle and apoptosis. Meanwhile, BET inhibitors (BETi) have regulatory effects on immune checkpoints, immune cells, and cytokine expression. The role of BET proteins and BETi in a variety of tumors has been studied. This paper reviews the recent research progress of BET and BETi in hematologic tumors (mainly leukemia, lymphoma and multiple myeloma) from cellular level studies, animal studies, clinical trials, drug combination, etc. BETi has a promising future in hematologic tumors, and future research directions may focus on the combination with other drugs to improve the efficacy.

1268

N6-methyladenosine (m⁶A) is the most abundant internal modification on RNA. It is a dynamical and reversible process, which is regulated by m⁶A methyltransferase and m⁶A demethylase. The m⁶A modified RNA can be specifically recognized by the m⁶A reader, leading to RNA splicing, maturation, degradation or translation. The abnormality of m⁶A RNA modification is closely related to a variety of biological processes, especially the occurrence and development of tumors. Recent studies have shown that m⁶A RNA modification is involved in the pathogenesis of skin cancers. However, the precise molecular mechanisms of m⁶A-mediated cutaneous tumorigenesis have not been fully elucidated. Therefore, this review will summarize the biological characteristics of m⁶A modification, its regulatory role and mechanism in skin cancers, and the recent research progress of m⁶A-related molecular drugs, aiming to provide new ideas for clinical diagnosis and targeted therapy of cutaneous cancers.

1330

De novo nucleotide biosynthetic pathway is a highly conserved and essential biochemical pathway in almost all organisms. Both purine nucleotides and pyrimidine nucleotides are necessary for cell metabolism and proliferation. Thus, the dysregulation of the de novo nucleotide biosynthetic pathway contributes to the development of many human diseases, such as cancer. It has been shown that many enzymes in this pathway are overactivated in different cancers. In this review, we summarize and update the current knowledge on the de novo nucleotide biosynthetic pathway, regulatory mechanisms, its role in tumorigenesis, and potential targeting opportunities.

1280

Pyroptosis is a regulated cell death pathway involved in numerous human diseases, especially malignant tumors. Recent studies have identified multiple pyroptosis-associated signaling molecules, like caspases, gasdermin family and inflammasomes. In addition, increasing in vitro and in vivo studies have shown the significant linkage between pyroptosis and immune regulation of cancers. Pyroptosis-associated biomarkers regulate the infiltration of tumor immune cells, such as CD4⁺ and CD8⁺ T cells, thus strengthening the sensitivity to therapeutic strategies. In this review, we explained the relationship between pyroptosis and cancer immunology and focused on the significance of pyroptosis in immune regulation. We also proposed the future application of pyroptosis-associated biomarkers in basic research and clinical practices to address malignant behaviors. Exploration of the underlying mechanisms and biological functions of pyroptosis is critical for immune response and cancer immunotherapy.

1434

Obesity has become a major health crisis in the past w50 years. The fat mass and obesity-associated (FTO) gene, identified by genome-wide association studies (GWAS), was first reported to be positively associated with obesity in humans. Mice with more copies of the FTO gene were observed to be obese, while loss of the gene in mice was found to protect from obesity. Later, FTO was found to encode an m⁶A RNA demethylase and has a profound effect on many biological and metabolic processes. In this review, we first summarize recent studies that demonstrate the critical roles and regulatory mechanisms of FTO in obesity and metabolic disease. Second, we discuss the ongoing debates concerning the association between FTO polymorphisms and obesity. Third, since several small molecule drugs and micronutrients have been found to regulate metabolic homeostasis through controlling the expression or activity of FTO, we highlight the broad potential of targeting FTO for obesity treatment. Improving our understanding of FTO and the underlying mechanisms may provide new approaches for treating obesity and metabolic diseases.

1899

Over the past few decades, advances in immunological knowledge have led to the identification of novel immune checkpoints, reinvigorating cancer immunotherapy. Immunotherapy, represented by immune checkpoint inhibitors, has become the leader in the precision treatment of cancer, bringing a new dawn to the treatment of most cancer patients. Galectin-9 (LGALS9), a member of the galectin family, is a widely expressed protein involved in immune regulation and tumor pathogenesis, and affects the prognosis of various types of cancer. Galectin-9 regulates immune homeostasis and tumor cell survival through its interaction with its receptor Tim-3. In the review, based on a brief description of the signaling mechanisms and immunomodulatory activities of galectin-9 and Tim-3, we summarize the targeted expression patterns of galectin-9 in a variety of malignancies and the promising mechanisms of antigalectin-9 therapy in stimulating anti-tumor immune responses.

1365

Protein arginine methylation is a common post-translational modification involved in the regulation of various cellular functions. Coactivator-associated arginine methyltransferase 1 (CARM1) is a protein arginine methyltransferase that asymmetrically dimethylates histone H3 and non-histone proteins to regulate gene transcription. CARM1 has been found to play important roles in cell differentiation and development, cell cycle progression, autophagy, metabolism, pre-mRNA splicing and transportation, and DNA replication. In this review, we describe the molecular characteristics of CARM1 and summarize its roles in the regulation of cell differentiation and development in mammals.

1413

Non-coding RNAs (ncRNAs) participate in the regulation of several cellular processes including transcription, RNA processing and genome rearrangement. The aberrant expression of ncRNAs is associated with several pathological conditions. In this review, we focused on recent information to elucidate the role of various regulatory ncRNAs i.e., micro RNAs (miRNAs), circular RNAs (circRNAs) and long-chain non-coding RNAs (lncRNAs), in metabolic diseases, e.g., obesity, diabetes mellitus (DM), cardiovascular diseases (CVD) and metabolic syndrome (MetS). The mechanisms by which ncRNAs participated in disease pathophysiology were also highlighted. miRNAs regulate the expression of genes at transcriptional and translational levels.circRNAs modulate the regulation of gene expression via miRNA sponging activity, interacting with RNA binding protein and polymerase II transcription regulation.lncRNAs regulate the expression of genes by acting as a protein decoy, miRNA sponging, miRNA host gene, binding to miRNA response elements (MRE) and the recruitment of transcriptional element or chromatin modifiers. We examined the role of ncRNAs in the disease pathogenesis and their potential role as molecular markers for diagnosis, prognosis and therapeutic targets. We showed the involvement of ncRNAs in the onset of obesity and its progression to MetS and CVD. miRNA-192, miRNA-122, and miRNA-221 were dysregulated in all these metabolic diseases. Other ncRNAs, implicated in at least three diseases include miRNA-15a, miRNA-26, miRNA-27a, miRNA-320, and miRNA-375. Dysregulation of ncRNAs increased the risk of development of DM and MetS and its progression to CVD in obese individuals. Hence, these molecules are potential targets to arrest or delay the progression of metabolic diseases.

1896

The ongoing global pandemic of coronavirus disease 2019 (COVID-19), caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has resulted in over 570 million infections and 6 million deaths worldwide. Early detection and quarantine are essential to arrest the spread of the highly contagious COVID-19. High-risk groups, such as older adults and individuals with comorbidities, can present severe symptoms, including pyrexia, pertussis, and acute respiratory distress syndrome, on SARS-CoV-2 infection that can prove fatal, demonstrating a clear need for high-throughput and sensitive platforms to detect and eliminate SARS-CoV-2.CRISPR-Cas13, an emerging CRISPR system targeting RNA with high specificity and efficiency, has recently drawn much attention for COVID-19 diagnosis and treatment. Here, we summarized the current research progress on CRISPR-Cas13 in COVID-19 diagnosis and treatment and highlight the challenges and future research directions of CRISPR-Cas13 for effectively counteracting COVID-19.

2070

Dendrites are specialized neuronal compartments that sense, integrate and transfer information in the neural network. Their development is tightly controlled and abnormal dendrite morphogenesis is strongly linked to neurological disorders. While dendritic morphology ranges from relatively simple to extremely complex for a specified neuron, either requires a functional secretory pathway to continually replenish proteins and lipids to meet dendritic growth demands. The Golgi apparatus occupies the center of the secretory pathway and is regulating posttranslational modifications, sorting, transport, and signal transduction, as well as acting as a non-centrosomal microtubule organization center. The neuronal Golgi apparatus shares common features with Golgi in other eukaryotic cell types but also forms distinct structures known as Golgi outposts that specifically localize in dendrites. However, the organization and function of Golgi in dendrite development and its impact on neurological disorders is just emerging and so far lacks a systematic summary. We describe the organization of the Golgi apparatus in neurons, review the current understanding of Golgi function in dendritic morphogenesis, and discuss the current challenges and future directions.

1354

Stem cell senescence and exhaustion, a hallmark of aging, lead to declines in tissue repair and regeneration in aged individuals. Emerging evidence has revealed that epigenetic regulation plays critical roles in the self-renew, lineage-commitment, survival, and function of stem cells. Moreover, epigenetic alterations are considered important drivers of stem cell dysfunction during aging. In this review, we focused on current knowledge of the histone modifications in the aging of mesenchymal stem cells (MSCs). The aberrant epigenetic modifications on histones, including methylation and acetylation, have been found in aging MSCs. By disturbing the expression of specific genes, these epigenetic modifications affect the self-renew, survival, and differentiation of MSCs. A set of epigenetic enzymes that write or erase these modifications are critical in regulating the aging of MSCs. Furthermore, we discussed the rejuvenation strategies based on epigenetics to prevent stem cell aging and/or rejuvenate senescent MSCs.

1307

Atherosclerosis is one of the leading causes of disease and death worldwide. The identification of new therapeutic targets and agents is critical. JAZF1 is expressed in many tissues and is found at particularly high levels in adipose tissue (AT). JAZF1 suppresses inflammation (including IL-1b, IL-4, IL-6, IL-8, IL-10, TNFa, IFN-g, IAR-20, COL3A1, laminin, and MCP-1) by reducing NF-κB pathway activation and AT immune cell infiltration. JAZF1 reduces lipid accumulation by regulating the liver X receptor response element (LXRE) of the SREBP-1c promoter, the cAMP-response element (CRE) of HMGCR, and the TR4 axis. LXRE and CRE sites are present in many cytokine and lipid metabolism gene promoters, which suggests that JAZF1 regulates these genes through these sites. NF-κB is the center of the JAZF1-mediated inhibition of the inflammatory response. JAZF1 suppresses NF-κB expression by suppressing TAK1 expression. Interestingly, TAK1 inhibition also decreases lipid accumulation. A dual-targeting strategy of NF-κB and TAK1 could inhibit both inflammation and lipid accumulation. Dual-target compounds (including prodrugs) 1-5 exhibit nanomolar inhibition by targeting NF-κB and TAK1, EGFR, or COX-2. However, the NF-κB suppressing activity of these compounds is relatively low (IC₅₀ > 300 nM). Compounds 6-14 suppress NF-κB expression with IC₅₀ values ranging from 1.8 nM to 38.6 nM.HS-276 is a highly selective, orally bioavailable TAK1 inhibitor. Combined structural modifications of compounds using a prodrug strategy may enhance NF-κB inhibition. This review focused on the role and mechanism of JAZF1 in inflammation and lipid accumulation for the identification of new anti-atherosclerotic targets.

1387

Lysine succinylation is a naturally occurring post-translational modification (PTM) that regulates the stability and function of proteins. It can be regulated by enzymes such as SIRT5 and SIRT7. Recently, the effect and significance of lysine succinylation in cancer and its implication in immunity have been extensively explored. Lysine succinylation is involved in the malignant phenotype of cancer cells. Abnormal regulation of lysine succinylation occurs in different cancers, and inhibitors targeting lysine succinylation regulatory enzymes can be used as potential anti-cancer strategies. Therefore, this review focused on the target protein lysine succinylation and its functions in cancer and immunity, in order to provide a reference for finding more potential clinical cancer targets in the future.

1380

Post-translational modifications (PTM) are covalent modifications of proteins or peptides caused by proteolytic cleavage or the attachment of moieties to one or more amino acids. PTMs play essential roles in biological function and regulation and have been linked with several diseases. Modifications of protein acylation (Kac), a type of PTM, are known to induce epigenetic regulatory processes that promote various diseases. Thus, an increasing number of studies focusing on acylation modifications are being undertaken. Butyrylation (Kbu) is a new acylation process found in animals and plants. Kbu has been recently linked to the onset and progression of several diseases, such as cancer, cardiovascular diseases, diabetes, and vascular dementia. Moreover, the mode of action of certain drugs used in the treatment of lymphoma and colon cancer is based on the regulation of butyrylation levels, suggesting that butyrylation may play a therapeutic role in these diseases. In addition, butyrylation is also commonly involved in rice gene expression and thus plays an important role in the growth, development, and metabolism of rice. The tools and analytical methods that could be utilized for the prediction and detection of lysine butyrylation have also been investigated. This study reviews the potential role of histone Kbu, as well as the mechanisms underlying this process. It also summarizes various enzymes and analytical methods associated with Kbu, with the goal of providing new insights into the role of Kbu in gene regulation and diseases.

1660

Long noncoding RNAs (lncRNAs) have been confirmed to play a crucial role in various biological processes across several species. Though many efforts have been devoted to the expansion of the lncRNAs landscape, much about lncRNAs is still unknown due to their great complexity. The development of high-throughput technologies and the constantly improved bioinformatic methods have resulted in a rapid expansion of lncRNA research and relevant databases. In this review, we introduced genome-wide research of lncRNAs in three parts: (i) novel lncRNA identification by high-throughput sequencing and computational pipelines; (ii) functional characterization of lncRNAs by expression atlas profiling, genome-scale screening, and the research of cancer-related lncRNAs; (iii) mechanism research by large-scale experimental technologies and computational analysis. Besides, primary experimental methods and bioinformatic pipelines related to these three parts are summarized. This review aimed to provide a comprehensive and systemic overview of lncRNA genome-wide research strategies and indicate a genome-wide lncRNA research system.

1365

Phosphatase and tensin homolog (PTEN) is a tumor suppressor gene and has a role in inhibiting the oncogenic AKT signaling pathway by dephosphorylating phosphatidylinositol 3, 4, 5-triphosphate (PIP₃) into phosphatidylinositol 4, 5-bisphosphate (PIP₂). The function of PTEN is regulated by different mechanisms and inactive PTEN results in aggressive tumor phenotype and tumorigenesis. Identifying targeted therapies for inactive tumor suppressor genes such as PTEN has been challenging as it is difficult to restore the tumor suppressor functions. Therefore, focusing on the downstream signaling pathways to discover a targeted therapy for inactive tumor suppressor genes has highlighted the importance of synthetic lethality studies. This review focused on the potential synthetic lethality genes discovered in PTEN-inactive cancer types. These discovered genes could be potential targeted therapies for PTEN-inactive cancer types and may improve the treatment response rates for aggressive types of cancer.

1398

Bone defects caused by diseases or surgery are a common clinical problem. Researchers are devoted to finding biological mechanisms that accelerate bone defect repair, which is a complex and continuous process controlled by many factors. As members of transcriptional costimulatory molecules, Yes-associated protein (YAP) and transcriptional co-activator with PDZ-binding motif (TAZ) play an important regulatory role in osteogenesis, and they affect cell function by regulating the expression of osteogenic genes in osteogenesis-related cells. Macrophages are an important group of cells whose function is regulated by YAP/TAZ. Currently, the relationship between YAP/TAZ and macrophage polarization has attracted increasing attention. In bone tissue, YAP/TAZ can realize diverse osteogenic regulation by mediating macrophage polarization. Macrophages polarize into M1 and M2 phenotypes under different stimuli. M1 macrophages dominate the inflammatory response by releasing a number of inflammatory mediators in the early phase of bone defect repair, while massive aggregation of M2 macrophages is beneficial for inflammation resolution and tissue repair, as they secrete many anti-inflammatory and osteogenesis-related cytokines. The mechanism of YAP/TAZ-mediated macrophage polarization during osteogenesis warrants further study and it is likely to be a promising strategy for bone defect repair. In this article, we review the effect of Hippo-YAP/TAZ signaling and macrophage polarization on bone defect repair, and highlight the regulation of macrophage polarization by YAP/TAZ.

1384

The retinal pigment epithelium (RPE) and choroid are located behind the human retina and have multiple functions in the human visual system. Knowledge of the RPE and choroid cells and their gene expression profiles are fundamental for understanding retinal disease mechanisms and therapeutic strategies. Here, we sequenced the RNA of about 0.3 million single cells from human RPE and choroids across two regions and seven ages, revealing regional and age differences within the human RPE and choroid. Cellecell interactions highlight the broad connectivity networks between the RPE and different choroid cell types. Moreover, the transcription factors and their target genes change during aging. The coding of somatic variations increases during aging in the human RPE and choroid at the single-cell level. Moreover, we identified ELN as a candidate for improving RPE degeneration and choroidal structure during aging. The mapping of the molecular architecture of the human RPE and choroid improves our understanding of the human vision support system and offers potential insights into the intervention targets for retinal diseases.

1353

Hand-foot syndrome (HFS) is a widely recognized dose-limiting cutaneous toxicity effect of fluoropyrimidine chemotherapy agents that impairs clinical benefits and treatment outcomes. Even though the cause and pathophysiology of HFS are relatively widely reported, how the toxicity of fluoropyrimidine translates into persistent inflammation has not been studied. Additionally, prevention and treatment strategies for HFS based on its mechanistic occurrence and development are scarce. In our study, we demonstrated that cGAS-STING signaling pathway-mediated cellular senescence played a critical role in the inflammatory reaction and provided a therapeutic solution for HFS. Mechanistically, DNA damage, as the primary cytotoxic cause, in keratinocytes induces cell cycle arrest, activates the cGAS-STING signaling pathway, and subsequently mediates cellular senescence, ultimately fueling a robust secondary inflammatory response that results in HFS. More importantly, the thymidine prodrug thymidine diacetate was proven to be effective in preventing HFS by compensating for thymidylate deficiency to facilitate the replication and repair of DNA and thus causing the escape from cellular senescence. These data highlight the importance of DNA damage-mediated cellular senescence in the etiology of HFS and provide a potential therapeutic anchor point for fluoropyrimidine-induced HFS.

1342

Endoplasmic reticulum (ER) membrane protein complex (EMC) is required for the cotranslational insertion of newly synthesized multi-transmembrane proteins. Compromised EMC function in different cell types has been implicated in multiple diseases. Using inducible genetic mouse models, we revealed defects in retinal vascularization upon endothelial cell (EC) specific deletion of Emc1, the largest subunit of EMC. Loss of Emc1 in ECs led to reduced vascular progression and vascular density, diminished tip cell sprouts, and vascular leakage. We then performed an unbiased transcriptomic analysis on human retinal microvascular endothelial cells (HRECs) and revealed a pivotal role of EMC1 in the β-catenin signaling pathway. Further in-vitro and in-vivo experiments proved that loss of EMC1 led to compromised β-catenin signaling activity through reduced expression of Wnt receptor FZD4, which could be restored by lithium chloride (LiCl) treatment. Driven by these findings, we screened genomic DNA samples from familial exudative vitreoretinopathy (FEVR) patients and identified one heterozygous variant in EMC1 that co-segregated with FEVR phenotype in the family. In-vitro expression experiments revealed that this variant allele failed to facilitate the expression of FZD4 on the plasma membrane and activate the β-catenin signaling pathway, which might be a main cause of FEVR. In conclusion, our findings reveal that variants in EMC1 gene cause compromised β-catenin signaling activity, which may be associated with the pathogenesis of FEVR.

1611

Bladder cancer (BLCA) remains a difficult malignancy to manage because of its high recurrence, intense follow-up, and invasive diagnostic and treatment techniques. Immune checkpoint inhibitors (ICIs) have forged a new direction for the treatment of BLCA, but it is currently challenging to predict whether an individual patient will be sensitive to ICIs. We collected 43 urine/tumor samples from BLCA patients for primary bladder cancer cells (BCCs) culturing using our previously reported BCC culture platform. We used flow cytometry (FCM) to measure the expression levels of Programmed Death-Ligand 1 (PD-L1) on BCCs before and after interferon-gamma (IFN-g) treatment and found that PD-L1 expression and the sensitivities to IFN-g varied among patients. RNA-sequencing, western blotting, and programmed death-1 (PD-1) binding assays confirmed that the BCC FCM-based PD-L1 detection platform (BC-PD-L1) was reliable and was not hindered by the glycosylation of PD-L1. In the subsequent retrospective study, we found that IFN-g-stimulated PD-L1 (sPD-L1) expression on BCCs detected by BC-PD-L1 could predict the prognosis of BLCA patients. Importantly, the prognostic value was similar or even better in urine-derived BC-PD-L1 (UBC-PD-L1). Transcriptome analysis showed that BCCs with high sPD-L1 tended to enrich genes associated with the collagen-containing extracellular matrix, cellecell adhesion, and positive regulation of the immune system. In addition, the UBC-PD-L1 also exhibited predictive value for ICI response in BLCA patients. In conclusion, as a novel personalized urine-detection method, UBC-PD-L1 may provide a rapid, accurate, and non-invasive tool for monitoring tumor progression, predicting therapeutic responses, and helping improve BLCA clinical treatment in future.

1354

To uncover the role of satellite cells (SCs) in paravertebral muscle development and aging, we constructed a single-nucleus transcriptomic atlas of mouse paravertebral muscle across seven timepoints spanning the embryo (day 16.5) to old (month 24) stages. Eight cell types, including SCs, fast muscle cells, and slow muscle cells, were identified. An energy metabolism-related gene set, TCA CYCLE IN SENESCENCE, was enriched in SCs. Forty-two skeletal muscle disease-related genes were highly expressed in SCs and exhibited similar expression patterns. Among them, Pdha1 was the core gene in the TCA CYCLE IN SENESCENCE; Pgam2, Sod1, and Suclg1 are transcription factors closely associated with skeletal muscle energy metabolism. Transcription factor enrichment analysis of the 42 genes revealed that Myod1 and Mef2a were also highly expressed in SCs, which regulated Pdha1 expression and were associated with skeletal muscle development. These findings hint that energy metabolism may be pivotal in SCs development and aging. Three ligand-receptor pairs of extracellular matrix (ECM) -receptor interactions, Lamc1-Dag1, Lama2-Dag1, and Hspg2-Dag1, may play a vital role in SCs interactions with slow/fast muscle cells and SCs self-renewal. Finally, we built the first database of a skeletal muscle single-cell transcriptome, the Musculoskeletal Cell Atlas (http: //www.mskca.tech), which lists 630, 040 skeletal muscle cells and provides interactive visualization, a useful resource for revealing skeletal muscle cellular heterogeneity during development and aging. Our study could provide new targets and ideas for developing drugs to inhibit skeletal muscle aging and treat skeletal muscle diseases.

1594

Gastric carcinoma (GC) progression is mainly caused by local aggression and lymph node metastasis. However, some patients with early T-stage disease have lymph node metastasis, whereas some patients with late T-stage disease do not have lymph node metastasis, which indicates that invasion and metastasis are not always sequential in some GC patients. In the present study, the data of 101 GC cases were acquired from TCGA and divided into T-late-N-negative and T-early-N-positive groups according to pathological stages. A total of 338 genes were identified as differential genes between the T-late-N-negative and T-early-N-positive groups. GSEA showed that epithelial cell signaling in the Helicobacter pylori (HP) infection pathway was enriched in the T-early-N-positive group. MB staining indicated that the HP infection rate was 63% (39/62) in N-positive patients compared to 42% (16/38) in N-negative patients. To investigate the potential mechanism, we focused on the gene chemokine (C-X-C motif) receptor 2 (CXCR2), which was not only clustered in the gene set of epithelial cells signaling in the HP infection pathway but also significantly upregulated in T-early-N-positive GC by the analysis of the different genes based on the TCGA dataset. A meta-analysis showed that CXCR2 expression was positively correlated with N-stage but not with T-stage in GC. This study indicated that invasion and metastasis could be independent processes driven by different molecular mechanisms in some GC patients. HP infection was a potential factor that promoted lymph node metastasis by upregulating CXCR2 expression.

1361

Protein arginine methyltransferase 1 (PRMT1), a type I PRMT, is overexpressed in gastric cancer (GC) cells. To elucidate the function of PRMT1 in GC, PRMT1 expression in HGC-27 and MKN-45 cells was knocked down by short hairpin RNA (shRNA) or inhibited by PRMT1 inhibitors (AMI-1 or DCLX069), which resulted in inhibition of GC cell proliferation, migration, invasion, and tumorigenesis in vitro and in vivo.MLX-interacting protein (MLXIP) and Kinectin 1 (KTN1) were identified as PRMT1-binding proteins. PRMT1 recruited MLXIP to the promoter of β-catenin, which induced β-catenin transcription and activated the β-catenin signaling pathway, promoting GC cell migration and metastasis. Furthermore, KTN1 inhibited the K48-linked ubiquitination of PRMT1 by decreasing the interaction between TRIM48 and PRMT1. Collectively, our findings reveal a mechanism by which PRMT1 promotes cell proliferation and metastasis mediated by the β-catenin signaling pathway.

1494