第9卷，第3期

Abdominal aortic aneurysm (AAA) is a common degenerative vascular disease with a prevalence up to 8% in males over 60 years of age. This is a complex disorder characterized by permanent and focal dilation of abdominal aorta that exceeds at least 50% of the normal diameter. In these patients, the aortic diameter progressively expands boosted by smoking and aging as major risk factors. Unfortunately, aneurysm growth increases the risk of aortic rupture, a life-threatening emergency that carries a mortality rate of 80%. Despite the increasing effort of the scientific community to identify therapeutic strategies for AAA, there are currently no pharmacological tools that ameliorate aneurysm expansion, while the underlying mechanisms involved in this disease are not completely understood.

1173

Tensin-1 (TNS1) is a 220 kD focal adhesion protein that binds to actin filaments, integrin receptors, small GTPases, tyrosine phosphorylated proteins, and lipids. These binding activities enable TNS1 to link the actin cytoskeleton to integrins and transduce outside-in and inside-out signals at focal adhesion sites, thereby regulating cell attachment, migration, proliferation, and mechanical sensing. Lack of TNS1 in kidney epithelial cells results in multiple lumen, instead of single lumen, phenotypes in the three-dimensional (3D) culture. Tns1 knockout (Tns1-KO) mice develop cystic kidneys and die from renal failure. These findings demonstrate critical roles of TNS1 inkidney epithelial cells and renal function. CTEN (aka TNS4) is a smaller 78 kD protein that only shares the SH2 and PTB domains with TNS1 and other tensin members. The expression patterns of TNS1, TNS2 and TNS3 in various human tissues are very similar and are frequently down-regulated in cancer samples. In contrast, CTEN expression is restricted to certain tissues, such as prostate and placenta, yet over-expressed in cancers including lung, colon, gastric, and breast cancers. It has been long speculated that CTEN might function as a dominant negative competitor of TNS1 based on its domain structure and expression pattern. However, this idea has not been convincingly demonstrated, especially in animal models.

1252

There are numerous online databases dedicated to genes associated with human diseases. The redundancy and lack of consistency of disease terms in these databases pose a significant impediment in high throughput data analysis. To address this, we developed GeDiPNet by compiling data available on gene-disease associations from six popular open access sources namely DisGeNET, ClinGen, ClinVar, HPO, OrphaNet, and PsyGeNET; and curated through several stages including manual curation and merging of disease names to ensure standard vocabulary (Supplementary file 1). GeDiPNet currently includes information on 12,270 genes that have disease association curated from literature records and 20,013 genes have disease association based on text mining.

1282

Preeclampsia (PE) is a pregnancy-specific hypertensive disorder which poses a severe threat to maternal and fetal health. Defective decidualization may contribute to PE. N6-methyladenosine (m6A) is associated with various diseases. The regulatory mechanism of m6A in PE is not well established to date. We aimed to identify differentially expressed m6A and explore its regulatory role in the pathogenesis of PE. Here we showed that m6A demethylase alkB homolog 5 (ALKBH5) was lower expressed in the decidua of PE. Silencing ALKBH5 suppressed the decidualization of human endometrial stromal cells (hESCs) and the invasion of trophoblast (HTR-8) cells into hESCs. The meRIP-seq analysis revealed the altered expression of ALKBH5 target genes after ALKBH5 knocking down, including CORIN. CORIN is a cardiac protease that activates atrial natriuretic peptide (ANP). Mechanism study indicated that ALKBH5 demethylated CORIN transcripts, maintained CORIN stability and enhanced CORIN expression through HuR dependent pathway, which was essential for ALKBH5 to regulate the decidualization of hESCs and the invasion of HTR-8 cells. In conclusion, our data indicate that low ALKBH5 expression in PE decidua inhibits the decidualization of hESCs and trophoblast cell invasion through the regulation of CORIN and ANP via the m6A-HuR-dependent pathway.

1238

Although the role of copper (Cu) in promoting KRas-or BRaf-mutation driven cancers via activating MEK1/2 kinases is known, the mechanism by which the copper transporter SLC31A1 (CTR1) is upregulated in pancreatic cancer (PDAC, KRas mutation) is not defined. In this study, we provide evidence that MEK signal maintains a high level of SLC31A1 through silencing the expression of miR-124-3P (miR-124) via a novel MEK-DNMT1-miR-124 feedback loop in PDAC cells. Further, we reveal that miR-124 directly targets suppression of SLC31A1, and miR-124 introduction together withtetrathiomolybdate (TM) treatmenthampered pancreatic cancer growth in vitro and in vivo. Our results demonstrate that a SLC31A1-MEK-DNMT1-miR-124 feedback loop is an important pathway to maintain copper absorption and promote pancreatic cancer progression, and we hope to provide a Cu-chelation as an adjuvant treatment strategy, to block the progression in Kras mutant PDAC patients.

1176

The schistosomiasis-associated colorectal cancer (SA-CRC) is thought to be caused by the chronic inflammation as a result of schistosomal ova deposition in the submucosal layer of the intestine. It is reported that SA-CRC differs from sporadic colorectal cancer (S-CRC) in terms of epidemiological features, histological patterns, and clinical outcomes, which is also supported by our institutional data. Here we investigate the genomic alternations and probable molecular determinants of pathogenesis in Chinese SA-CRC.

1218

Congenital cataract is a clinically and genetically heterogeneous disease characterized by any opacity of the lens presenting at birth or in the first year of life, with an incidence of 1-6/10,000 live births in developed countries and 5-15/10,000 live births in developing countries. Congenital cataract accounts for 7.4%-15.5% of all childhood blindness, and it occurs as either a syndromic disease or an isolated (nonsyndromic) disease with or without other ocular malformations, such as microcornea, microphthalmia, persistent fetal vascularization, glaucoma, or retinal dystrophies. Whereas autosomal dominant inheritance is most common, autosomal recessive and X-linked inheritance have also been reported, indicating a degree of genetic heterogeneity in congenital cataract. To date, various cataract-associated loci have been detected, with over 453 genes associated with cataracts reported prior to October 1,2021 (http://cat-map.wustl.edu/). The pathogenic variation detection rate in familial cases was approximately 75%, indicating that the genetic basis for approximately one-fourth of familial patients is still unknown. In recent years, there has been a shift from traditional genetictests (e.g., Sanger sequencing) to genomic tests (comparative genomic hybridization (CGH)), gene panels, and next-generation sequencing (NGS) technologies) for many diseases with complex inheritance and genotypes, especially rare Mendelian diseases. NGS technologies, such as whole-exome sequencing (WES) and wholegenome sequencing (WGS), are important for identifying pathogenic variations, especially for discovering new pathogenic genes in Mendelian diseases with genetic heterogeneity. With the development of NGS technologies, more disease-causing genes associated with congenital cataract will be discovered. In this study, we identified a novel association between a missense variation in EXD3 and congenital cataract in three Chinese families by combining NGS technologies and linkage analysis.

1225

The pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) continues to sweep the globe with devastating consequences on human lives and world economy. As an RNA virus, SARS-CoV-2 has a relatively high mutation rate and is rapidly evolving. Thus, new SARS-CoV-2 variants continued to emerge, 5 of which were designated by the World Health Organization (WHO) as variants of concern (VOCs), Alpha (B.1.1.7), Beta (B.1.351), Gamma (P.1), Delta (B.1.617.2), and, recently, Omicron (B.1.1.529). First identified in Botswana and South Africa in November 2021, the original Omicron, BA.1, then spread to every corner of the world and quickly replaced the previously dominant Delta strain to become the most prevalent SARS-CoV-2 circulating variant across the world. BA.1 is reported to escape most therapeutic monoclonal antibodies against SARS-CoV-2. Consistently, sera from convalescent donors and vaccinated individuals contain very low to undetectable levels of neutralizing antibodies against BA.1. Therefore, new therapeutic agents are urgently needed.

1194

SARS-CoV-2 is the causal agent of the COVID-19 pandemic. This is a single-stranded RNA beta-coronavirus that is comprised of a 5′ and 3′-untranslated region (UTR) in its non-coding RNA (ncRNA) region. The P-element-induced wimpy testis (PIWI) -interacting RNAs (piRNAs) are ncRNA sequences that bind to transposons and interfere with the translation of new genes. Here, we search for short sequences in the non-coding regions, corresponding to 28 nucleotides (nt). These start from a uracil and have a conserved region of at least 10 nt of homology with the previously reported piRNAs. The aim of this work was to identify piRNA-like sequences in the 3′-UTR of SARS-CoV-2 from the reported Wuhan Patient's genome sequence (GenBank: MN908947.3).

1168

Sudden cardiac death (SCD) is the leading cause of death from out-of-hospital cardiovascular disease worldwide, with an annual incidence of 1.4 per 100,000 in females and 6.8 per 100,000 in males. Most cases of SCD in older patients (>45 years) are caused by atherosclerotic coronary artery disease. SCD in younger patients (<45 years) is caused by inherited cardiovascular diseases, including premature atherosclerosis, primary electrical disease, cardiomyopathy, thoracic aortic aneurysm, and dissection. Primary electrical disease, caused by molecular defects in the ion channels involved in the generation of cardiac action potentials, accounts for approximately 93% of SCDs from a hereditary disease in patients with negative autopsy findings.

1196

Chemoresistance is still one of the main obstacles to treat colorectal cancer (CRC). Substantial studies on biomarkers related to chemoresistance have emerged in recent years. Many microRNAs (miRNA) have been reported to involved in drug resistance and CRC. MiR-126-5p participates in malignant behaviors of numerous cancers. However, it is not known if miR-126-5p participates in multidrug resistance in CRC. Especially, the impact of miR-126-5p on drug resistance of CRC and its drug resistance mechanism have not been reported.

1190

Fibroblast growth factor 23 (FGF23) is an osteocyte-and osteoblast-derived hormone that primarily regulates phosphate and vitamin D metabolism. Circulatory FGF23 levels are abnormally increased in pathological conditions like acute or chronic kidney injury, resulting in disease progression as well as increased rates of morbidity and mortality. However, FGF23 production in acute liver injury is not fully investigated. In this study, we report that carbon tetrachloride (CCl₄) -induced acute liver injury upregulates hepaticestrogen-relatedreceptorgamma (ERRγ) and FGF23 gene expression and FGF23 secretion from liver. Hepatocyte specific depletion of ERRγ bluntedCCl₄-induced hepatic FGF23 promoter activity, FGF23 gene expression and FGF23 levels. Further, treatment with ERRγ-specific inverse agonist GSK5182 also efficiently inhibited CCl₄-induced acute liver injury-mediated hepatic FGF23 gene expression and circulatory FGF23 levels in vivo. Taken together, these results firstly describe a detailed molecular mechanism of hepatic FGF23 gene expression induction in an acute liver injury condition. Further, we present evidence that inhibiting ERRγ transactivation by the small molecule GSK5182 may be a useful strategy to control the devastating circulatory levels of FGF23.

1501

Studies have shown that dysregulated expression of miRNAs is a major underlying event in the onset and progression of many metabolic diseases. The pathological role of miR-548c-3p in various cancer types, including breast cancer, glioma, prostate cancer, and gastric cancer, has been documented. However, experimental evidences validating the role of miR-548c-3p in the development of metabolic diseases are still lacking, and no data is available on its diagnostic potential for metabolic syndrome (MS). Recently, we predicted the involvement of miR-548c-3p in the pathogenesis of MS and its progression to cardiovascular diseases (CVD) via the regulation of target genes necessary for the proper function of insulin resistance, mTOR, and JAK/STAT signaling pathways. Therefore, we deemed it necessary to engage in wet laboratory analysis and validate these claims in patients with metabolic syndrome compared to healthy individuals.

1824

The discovery of clinically useful targets to treat specific pathological conditions can provide novel therapeutic approaches. To unbiasedly identify key genes related to tumor metastasis, we developed a random gene perturbation method using a piggyBac transposon system under the control of a doxycycline regulated promoter. Using this random gene perturbation method and utilizing a mouse model of metastatic pancreatic cancer, we identified genes dysregulated in metastasized cells from a random mutagenesis library after multiple rounds of in vivo selection. Analysis of these metastasized clones revealed the downregulation of ARPC1B gene. Our further mechanistic studies revealed that ARPC1B gene and its closely related gene ARPC1A worked in a regulatory loop to control tumor metastasis. These findings validate that piggyBac transposon mediated random gene perturbation is a powerful tool to investigate the functional relevance of novel genes and the ARPC1A/B axis is a potential key regulator of tumor metastasis.

1400

Chanarin Dorfman Syndrome (CDS; MIM: 275630) is a rare autosomal recessive disorder, characterized by tri-acylglycerol (TG) accumulation in lipid droplets (LDs) within different tissues including skin, liver, skeletal muscle, bone marrow, eyes, ears, and central nervous system. In CDS, the prevalent and always observed clinical feature is a nonbullous congenital ichthyosiform erythroderma (NCIE). Liver and muscle involvement is detected in more than 80% and 40% of cases, respectively. Neurological symptoms have been reported in almost 30% of affected subjects. The disease is due to mutations in the α/β-hydrolase domain-containing protein 5(ABHD5). This protein can activate both adipose triglyceridelipase (ATGL), thefirstrate-settingenzymeinTG hydrolysis, and PNPLA1, which catalyzes the final step of ὠ-O-acylceramide production.

1173

Fasting induces mammalian metabolic switch from glucose to fatty acid-derived ketones, resulting in a marked change in blood glucose, triglyceride, free fatty acid and bhydroxybutyrate levels and a phenotype of transient hepatic steatosis. However, the underlying mechanism remains incompletely understood. This study aimed to investigate the role of 3-hydroxy-3-methylglutaryl-CoA synthase 2 (HMGCS2), a rate-limiting enzyme in ketogenesis, in fasting-induced ketogenesis and hepatic lipid accumulation. Hepatocyte-specific HMGCS2 knockout (LKO) mice were generated and used to compare the difference in liver metabolic response to fasting between wild-type (WT) and LKO mice. Our findings demonstrate that HMGCS2 is essential in enhanced hepatic ketogenesis during fasting. Dysfunction of HMGCS2 leads to excessive hepatic triglyceride accumulation and severe liver injury possibly via increased liver fatty acid uptake by CD36. Therefore, liver HMGCS2 may represent a key enzyme in the maintenance of liver lipid homeostasis during fasting.

1215

Understanding DNA methylation (DNAm) alterations in tumors is important to develop novel therapeutic targets and powerful biomarkers, and to gain insights into tumorigenesis processes. As many studies have highlighted the confounding effects of tumor purity in DNAm analyses, various tools have also been developed to tackle the issue. Among them, InfiniumPurify R package is a popular and widely cited tool, consisting of a set of statistical methods to estimate and account for tumor purity in cancer DNAm analyses.

1238

Glycogen storage disease (GSD) is a rare autosomal recessive disease by abnormal accumulation of intracellular glycogen. Mutations in genes encoding G-6-P or G-6-Pase lead to dysfunction of the body's glycogen metabolism. Without adequate metabolic treatment, patients with GSD can die during infancy or childhood from severe hypoglycemia and acidosis. The patient's symptoms generally achieve remission during adolescence, except in rare cases when cirrhosis of the liver or myopathy occurs. Once GSD is identified in adults, the patients are often accompanied by many complications, such as fasting hypoglycemia, hyperlipidemia, hyperlactatemia and hyperuricemia. In most cases, these complications may mask the patient's primary disease. At present, there are more than 14 types of GSD. The details were presented in. It's worth noting that Glycogen storage disease type IA (GSD-IA) is the most prevalent subtype and represents approximately 80% of GSD-I cases. It's characterized by a deficiency of glucose-6 phosphatase (G-6-P), a key enzyme in glycogen metabolism. However, G-6-Pase deficiency blocked the normal pathway of glycogenolysis and glycogen is accumulated excessively in the liver or muscle. This is why the patients with GSD are typically characterized by hypoglycemia and hepatomegaly. Once the metabolic alternative pathways are activated, the body will develop various complications.

1294

One of the main reasons that hinders the application of base editors in the clinic is the trade-off between editing efficiency and editing fidelity. The off-target base editing includes Cas9-independent and Cas9-dependent manners. The Cas9-independent off-target results from the intrinsic deamination activity of the deaminase domain of a base editor, which affects random RNA or DNA molecules. Meanwhile, the Cas9-dependent off-target results from the binding of Cas9 domain to DNA sequences that show strong similarity to the target sequence. Compared to Cas9-indepedentoff-targetthathasbeenextensively studied, Cas9-dependentoff-targetbaseeditinghas received much less attention. In principle, Cas9-dependent off-target base editing could be avoided by using a high-fidelity Cas9 in the base editor. Quite a few high-f idelity Cas9 variants have been reported. Among them, several have been tested as a component of a base editor in mammalian cells. However, all these studies focused on cytosine base editor (CBE) with one recent exception in which Sniper-Cas9 is used in an adenosine base editor (Sniper ABE7.10) to reduce off-target editing. Hence, it is still unclear how high-fidelity Cas9 variants could improve the performance of ABE.

1318

Metabolic syndrome (MetS) is a complex condition that significantly increases the risk of several diseases, including heart disease and diabetes. Patients with MetS show both pro-inflammatory and pro-thrombotic states; however, the state that takes precedence in MetS progression is unclear. The molecular mechanisms underlying the prodromal features in MetS have still not been well-elucidated. In this cohort-based study, we analyzed the characteristics of pre-MetS by dividing 100 participants into three groups: normal, pre-MetS, and MetS. We performed a systematic transcriptome analysis using RNA-seq data obtained from the peripheralbloodofindividuals. Geneexpressionand pathway enrichment analyses revealed that immune-related dysregulation was prominent in the pre-MetS group. In contrast, the MetS group showed significant coagulationrelated dysregulation. These results suggest that the pro-inflammatory state may be a prodromal feature of pre-MetS, followed by a pro-thrombotic state during progression to MetS. Our findings may contribute to the prevention of MetS progression by encouraging the development of diagnostic and therapeutic strategies that target the identified pre-MetS-associated genes and pathways.

1851

Loss-of-function mutations of interleukin 10 receptor alpha subunit (IL-10RA) (homozygotes or compound heterozygotes) are associated with pediatric colitis and diarrhea. These mutations always drive a significant increase in serum interleukin-10 (IL-10) levels, while such high levels of IL-10 fail to suppress intestinal inflammation, at least in part by failure to suppress the release of some cytokines such as interleukin 1 beta (IL-1β) and tumor necrosis factoralpha (TNFα). However, at present, whether a novel missense variant has a pathogenic effect is mainly determined according to the guidelines issued by the American College of Medical Genetics and Genomics (ACMG). Without functional verification, a variant of uncertain significance (VUS) will be easily made, or a variant will be misjudged as pathogenic or benign. Thus, a function test of a novel mutated IL-10RA is needed to determine whether colitis was attributed to such a mutation.

1261

Colorectal cancer (CRC) is the third most common and lethal cancer worldwide. Farnesoid X receptor (FXR) is a regulator of bile acid (BA) homeostasis and its expression is inversely correlated with the incidence of CRC. Moreover, our recent study discovered that dietary factors and dysregulated Wnt signaling independently alter BA profiles, some of which antagonize intestinal FXR, such as Tauro-bmuricholic acid (T-β MCA) and deoxycholic acid (DCA). These antagonistic BAs drove malignant transformations in Lgr5-expressing (Lgr5⁺) intestinal stem cells (ISCs), which promoted an adenoma-to-adenocarcinoma progression. Conversely, FXR agonists restored BAs homeostasis and curtailed CRC progression. However, the precise mechanism of FXR inhibiting Wnt signaling remains unclear, prompting us to investigate FXR's targets in Wnt pathway, especially in ISCs.

1175

Since the end of 2019, COVID-19 has caused worldwide pandemic. SARS-CoV-2, the culprit of this epidemic, binds to the host receptor Angiotensin converting enzyme 2 (ACE2) using spike (S) protein for cell entry. Recent studies showed that ACE2 expressed at a low level in the main target organs such as lung and bronchial tissues. How SARS-CoV-2 efficiently invades into the respiratory system or other organs with low levels of ACE2 is an urgent problem to be solved. Here, we discovered that NSP16 significantly promoted the invasion of SARS-CoV-2 pseudovirus. NSP16 could promote the formation of a STUB1-USP14 de-ubiquitination complex that regulated the ubiquitination and stability of transmembrane serine protease 2 (TMPRSS2). Inhibiting the function of this complex remarkably reduced the invasion ability of SARS-CoV-2 pseudovirus. Thus, our study demonstrates for the first time that NSPs of SARS-CoV-2 can also participate in viral invasion.

1248

Female germline stem cells (FGSCs) and spermatogonial stem cells (SSCs) are germline stem cells that transmit the genetic material from generation to generation. Hence, it is essential to understand the similarities and differences between FGSCs and SSCs. In this study, we comprehensively compared the epigenetics of FGSCs and SSCs, including high-orderchromatinorganization, chromatinhistone modification, and transcriptome by analyzing multipleomics data (Hi-C, ChIP-seq, and RNA-seq). We found that FGSCs and SSCs had both similar and different chromatin histone modifications that were correlated with their chromatin organization. We also identified cell-specific enhancerepromoterinteractionsinFGSCsandSSCs. Furthermore, we found that the X chromosome was similar in FGSCs and SSCs by principal component analysis of the X chromosome. By using RNA fluorescence in situ hybridization (RNA FISH) to confirm the inactivation of the X chromosome in FGSCs, we compared the high-order chromatin structure between activated and inactivated X chromosomes in FGSCs. In summary, we performed a systematic comparison analysis of the epigenetics between FGSCs and SSCs from multiple perspectives using multiple-omics data.

1249

Heterotopic ossification (HO) describes bone formation at non-skeletal sites and results from traumatic injury, surgery, or genetic disease such as fibrodysplasia ossificans progressiva (FOP). Although it is known that BMP signaling regulates HO, knowledge about the developmental origin of the osteogenic progenitors responsible for the BMP-associated metamorphosis is comparably less. With the use of transgenic mice and labelled neural crest-derived cell, we found myelin protein zero (P0, or MPZ) -and Wnt1-lineage cells give rise to BMP-7 induced adult ectopic cartilage and bone. In addition, the induced expression of ACVR1 (R206H), which is the major mutation found in FOP patients, in P0-lineage cells formed ectopic bone after cardiotoxin-induced muscle injury. We also found that the majority of muscle-resident fibro-adipogenic progenitors (FAPs), essential for muscle homeostasis and responsible for HO in skeletal muscle, are derived from P0- and Wnt1-lineage cells. The data collectively suggest that muscle-resident neural crest-derived progenitor cells account for both nonhereditary and genetic type HO.

1276

Ferroptosis is a form of regulated cell death characterized by iron-dependent overaccumulation of lipid peroxides, which causes membrane damage and cell lysis. Ferroptosis can be prevented by cellular antioxidant mechanisms such as glutathione peroxidase 4 (GPX4)-mediated elimination of the lipid peroxides at the cost of glutathione (GSH). As the rate-limiting step of GSH synthesis, the availability of intracellular cystine is controlled by the cell membranelocated cystine/glutamate antiporter system x_c^-. Indeed, the initially identified small molecule inducers of ferroptosis, RSL3 and erastin, turned out to be inhibitors of GPX4 and system x_c^-, respectively.

1250

Of over 170 types of RNA modifications discovered, N⁶-methyladenosine (m⁶A) is the most actively studied. m⁶A confers oncogenic potential, maintains cancer stem cells, and is associated with cancer-related outcomes. The role of RNA modifications including m¹A is poorly understood. m¹A is abundant in transfer and ribosomal RNAs but has also been found on messenger RNAs. m¹A stabilizes these RNA species to regulate essential biological processes including translation and ribosome biogenesis, which are frequently dysregulated in cancers. However, the role of aberrant m¹A RNA methylation in cancer development and its utility in predicting clinical outcomes is currently unclear. Understanding the function of m¹A RNA methylation in diverse cancers requires the characterization of genetic alterations and aberrant expression of m¹A regulatory genes. These genes include m¹A writers (TRM61A, TRIM61B, TRIMT61B, TRMT10C), erasers (ALKBH1, ALKBH3) and readers (YTHDF1, YTHDF2, YTHDF3 and YTHDC1). Here, we present the frequency of molecular abnormalities affecting m¹A regulatory genes and their association with cancer-related pathways, prognosis, and tumor immune cell infiltration across 33 types of human cancers.

1312

Primary ciliary dyskinesia (PCD) is an autosomal recessive disease caused by defects in motile cilia and clinically characterized by bronchiectasis, situs inversus, nasosinusitis, recurrent respiratory infections, tympanitis, and/or male infertility. In PCD, impaired function or structure of motile cilia leads to abnormality of mucociliary clearance, and RSPH4A (encoding radial spoke head protein) mutation has been recognized as a major causative factor. Hereditary myopathy with early respiratory failure (HMERF) is an autosomal dominant myopathy due to mutations in TTN gene (encoding the fibronectin III domain of titin). HMERF is an extremely rare condition characterized by severe respiratory involvement at onset and muscle weakness starting in the third to fifth decades of life.

1266

Herpes simplex virus 1(HSV-1)is highly infectious. More than 70% of the world's population has been infected with HSV-1. Most patients with HSV-1 have no obvious clinical symptoms. However, HSV-1 infection can lead to serious complications in people with an underlying genetic susceptibility, various causes of immunodeficiency, or immunocompromised populations. HSV-1 has the property of activating periodically in neurons and is considered to be one of the risk factors for Alzheimer's disease (AD). However, the underlying pathological mechanisms causing the development of AD induced by HSV-1 are not yet well established. With the spread of transcriptome sequencing and advances in bioinformatics technology, we can gain a deeperunderstandingoftheunderlyingpathological mechanisms behind the occurrence of disease, which can help in clinical diagnosis and clinical treatment of diseases. In the present study, we used the Gene Expression Omnibus (GEO) database to obtain sequencing datasets related to HSV-1 and AD, explored the potential pathogenesis of HSV-1 leading to AD using various algorithms in bioinformatics, and combined with public databases to construct a potential competing endogenous RNAs (ceRNA) regulatory network for HSV-1 leading to AD.

1209

Tetralogy of Fallot (TOF) is the most common cyanotic congenital heart disease and the incidence of late cardiac death in long-term survivors continues to increase. So, there is an urgent need to explore the etiology and pathogenesis of TOF. The precise cause of TOF is currently unclear, and exploration of the pathogenesis has focused increasingly in recent years on the roles of noncoding gene products, especially long noncoding RNAs (lncRNAs). The lncRNA NRON is localized at the 9q33.3 revered strand. The related literature has revealed that its transcript is enriched in human cardiac muscle and NRON over-expression has been reported to reverse epithelial-to-mesenchymal transition (EMT) which is known to contribute to valve development in the heart. Based on these findings, we hypothesized that abnormal expression of NRON was probably relevant to congenital heart defects. This study investigated the role of lncRNA NRON in the nucleus and its correlation with cardiac development and TOF.

1223

Isabel Quintana，Mariona Terradas，Pilar Mur，Iris B.A.W. te Paske，Sophia Peters，Isabel Spier，Verena Steinke-Lange，Claudia Maestro，David Torrents，Montserrat Puiggro`s，Romina Royo，Raul Tonda，Genís Parra，Davide Piscia，Sergi Beltrán，Matilde Navarro，Virginia Pin˜ol，Joan Brunet，Noemi Gonzalez-Abuin，Gemma Aiza，Anna Sommer，Yasmijn van Herwaarden，Galuh Astuti，Elke Holinski-Feder，Nicoline Hoogerbrugge，Richarda M. de Voer，Stefan Aretz，Gabriel Capellá，Laura Valle

Much of the genetic predisposition to polyposis, and particularly to serrated polyposis (SP), remains unknown. Only germline pathogenic variants in RNF43, a tumor suppressor that exerts negative feedback in the Wnt/β-catenin signaling pathway, have been causally linked to some SP cases (<2%), a disease associated with increased risk of colorectal cancer (CRC). Most known hereditary CRC and polyposis genes affect DNA repair, BMP/TGF-b, or Wnt signaling, being the latter associated with adenomatous and serrated polyposis phenotypes. Based on this observation, we evaluated the presence and role of germline variants in those pathways in unsolved polyposis patients.

1230

The dual-specificity tyrosine-regulated kinase (DYRK) family is evolutionarily conserved from invertebrate to mammals. DYRKs regulate cell proliferation, apoptosis, survival, and differentiation by modifying the protein activation state, cellular localization, and turnover. In contrast to several studies in cellular models, the available evidence regarding the in vivo roles of DYRKs in mammalian development is limited. This review summarizes the in vivo studies on Dyrks which provide insight into their roles in mammalian tissue development and congenital diseases. In vivo evidence obtained using knockout and genetically modified animals helps to understand and develop novel clinical therapies and drug for human congenital diseases, such as Down syndrome and neuronal disorders (associated with DYRK1A) and skeletal ciliopathies (associated with DYRK2).

1297

Vascular remodeling and angiogenesis are two key processes in the maintenance of vascular homeostasis and involved in a wide array of vascular pathologies. Following these processes, extracellular matrix (ECM) provides the mechanical foundation for vascular walls. Lysyl oxidase (LOX), the key matrix-modifying enzyme, has been demonstrated to significantly affect structural abnormality and dysfunction in the blood vessels. The role of LOX in vascular remodeling and angiogenesis has always been the subject in the current medical research. Therefore, we presently make a summarization of the biosynthesis of LOX and the mechanisms involved in vascular remodeling and angiogenesis, as well as the role of LOX in diseases associated with vascular abnormalities and the therapeutic potential via targeting LOX. In particular, we give a proposal that LOX likely reshapes matrisome-associated genes expressions in the regulation of vascular remodeling and angiogenesis, which serves as a mechanistic insight into the critical role of LOX in these two aspects. Additionally, LOX has also dual effects on the vascular dysfunction, namely, inhibition of LOX for improving hypertension, restenosis and malignant tumor while activation of LOX for curing arterial aneurysm and dissection. LOX-targeted therapy may provide a promising therapeutic strategy for the treatment of various vascular pathologies associated with vascular remodeling and angiogenesis.

1275

Parkinson's disease (PD) is the most common neurodegenerative movement disorder in the elderly. As the pathogenesis of PD is still not fully understood, medications with the capacity of halting the disease progression are currently unavailable. The discovery of genes that are causative for, or increase susceptibility to PD is pivotal for the development of novel therapeutic approaches, as they are critical for the onset of PD and the molecular pathways underlying its pathogenesis. By reviewing relevant data, we discuss causative genes, and those associated with PD susceptibility and quantitative traits. Through Gene Ontology database and STRING analysis, we emphasize the roles of inorganic cation transmembrane transport pathways and hypothalamic pituitary thyroid axis, in addition to the established roles of inflammation/oxidative stress and mitochondrial dysfunction in the pathogenesis of PD. It is hoped these insights 1) untangle the clinical complex presentations of PD, 2) reveal the inter-woven molecular network leading to PD, and 3) identify critical molecular targets to facilitate novel PD drug discovery, with a view to providing improved consultation and personalized medicine for patients with PD in the future.

1318

Obesity plays a crucial role in the development of non-alcoholic fatty liver disease (NAFLD). However, the underlying mechanism for the pathogenesis of obesity-associated NAFLD remains largely obscure. Although the "multiple hit" theory provides a more accurate explanation of NAFLD pathogenesis, it still cannot fully explain precisely how obesity causes NAFLD. The liver is the key integrator of the body's energy needs, receiving input from multiple metabolically active organs. Thus, recent studies have advocated the "multiple crosstalk" hypothesis, highlighting that obesity-related hepatic steatosis may be the result of dysregulated "crosstalk" among multiple extra-hepatic organs and the liver in obesity. A wide variety of circulating endocrine hormones work together to orchestrate this "crosstalk". Of note, with deepening understanding of the endocrine system, the perception of hormones has gradually risen from the narrow sense (i. e. traditional hormones) to the broad sense of hormones as organokines and exosomes. In this review, we focus on the perspective of organic endocrine hormones (organokines) and molecular endocrine hormones (exosomes), summarizingsystematically how the two types of new hormones mediate the dialogue between extrahepatic organs and liver in the pathogenesis of obesity-related NAFLD.

1261

AT-rich interactive domain 5a (Arid5a) is a member of the arid family of proteins, which contain a helix-turn-helix domain and an ability to bind to nucleic acids. Current evidence suggests that Arid5a performs dual functions as a transcription factor and an RNA-binding protein in immune, nonimmune, and/or tumor cells depending on its cellular localization. The contribution of Arid5a to the development of inflammation, autoimmunity, and obesity through its transcriptional and posttranscriptional regulatory functions has broadly been reviewed. Recent studies have indeed revealed an association of Arid5a with cancers, including breast, pancreatic, colorectal, and lung cancers and glioma. Notably, Arid5a affects various aspects of cellular homeostasis, including invasion, metastasis, epithelial-to-mesenchymal transition, immune evasion, adipogenesis and M1-like tumor-associated macrophage (TAM) -to-M2-like TAM transition. This review aims to summarize current knowledge of Arid5a from a cancer perspective and highlights recent advances in Arid5a-related cancer research. This review may improve the understanding of Arid5a-mediated molecular mechanisms and their relevance to cancers.

1314

Due to excess energy intake and a sedentary lifestyle, the prevalence of obesity is rising steadily and has emerged as a global public health problem. Adipose tissue undergoes structural remodeling and dysfunction in the obese state. Secreted proteins derived from the liver, also termed as hepatokines, exert multiple effects on adipose tissue remodeling and the development of obesity, and has drawn extensive attention for their therapeutic potential in the treatment of obesity and related diseases. Several novel hepatokines and their functions on systemic metabolism have been interrogated recently as well. The drug development programs targeting hepatokines also have shown inspiring benefits in obesity treatment. In this review, we outline how adipose tissue changes during obesity. Then, we summarize and critically analyze the novel findings on the effects of metabolic "beneficial" and metabolic "harmful" hepatokines to adipose tissue. We also discuss the in-depth molecular mechanism that hepatokines may mediate the liver-adipose tissue crosstalk, the novel technologies targeting hepatokines and their receptors in vivo to explore their functions, and the potential application of these interventions in clinical practice.

1320

As members of the immune checkpoint family, PD-1 and its ligand PD-L1 play critical roles in maintaining the balance between autoimmunity and tolerance. The interaction of PD-1/PD-L1 is also involved in tumor evasion inside the tumor microenvironment, caused by reduced T cell activation, proliferation, cytotoxic secretion, and survival. Previous research has shown that the expression level of PD-1/PD-L1 may be regulated by ubiquitin-mediated proteasome degradation, which is an important mode of post-translational modification (PTM). PD-1/PD-L1 ubiquitin modification research in tumor immunotherapy is the subject of the present review, which aims to assess the most recent developments in this area. We offer a short explanation of PD-1/PD-L1 as well as some basic background information on the UPS system and discuss many routes that target E3s and DUBs, respectively, in the regulation of PD-1/PD-L1 in tumor immunotherapy. In addition, we offer numerous innovative prospective research areas for the future, as well as novel immunotherapy concepts and ideas. Taken together, the information compiled herein should serve as a comprehensive repository of information about tumor immunotherapy that is currently available, and it should be useful in the design of future studies, as well as the development of potential targets and strategies for future tumor immunotherapy.

1315

The skeletal system is a dynamically balanced system, which undergoes continuous bone resorption and formation to maintain bone matrix homeostasis. As an important ADP-ribosylase and NAD⁺-dependent deacylase, SIRT6 (SIR2-like protein 6) is widely expressed on various kinds of bone cells, such as chondrocytes, osteoblasts, osteoclasts. The aberration of SIRT6 impairs gene expression (e.g., NF-kB and Wnt target genes) and cellular functions (e.g., DNA repair, glucose and lipid metabolism, telomeric maintenance), which disturbs the dynamic balance and ultimately leads to several bone-related diseases. In this review, we summarize the critical roles of SIRT6 in the onset and progression of bone-related diseases including osteoporosis, osteoarthritis, rheumatoid arthritis, and intervertebral disc degeneration, as well as the relevant signaling pathways. In addition, we discuss the advances in the development of SIRT6 activators and elucidate their pharmacological profiles, which may provide novel treatment strategies for these skeletal diseases.

1256

Cysteine dioxygenase type 1 (CDO1), belonging to the mammalian non-heme Fe (II) dioxygenases family, is a key enzyme for cysteine catabolism. Its activity and expression is regulated through multiple mechanisms. CDO1 is involved in a spectrum of physiological processes including lipid metabolism, adipogenesis, osteoblastic differentiation, redox homeostasis, fertility, bile acid metabolism, sulfide metabolism, and organismal growth and development. Many of these processes are regulated directly or indirectly by CDO1-mediated metabolism of cysteine. In pathophysiological processes, the degree of CDO1 promoter methylation is closely related to the progression and malignancy of tumors, and overexpression of CDO1 will promote ferroptosis of cancer cells. Moreover, CDO1 may ameliorate metabolic disorders through the taurine-mediated improvement of lipid metabolism and insulin sensitivity and improve neurodegenerative diseases by regulating cysteine level. Therefore, elucidation of the mechanisms underlying the role of CDO1 would provide a clearer view of the therapeutic potential and possible risks of targeting this important enzyme.

1236

Sterile a motif and histidine/aspartic acid domain containing protein 1 (SAMHD1) is a deoxynucleoside triphosphate (dNTPs) triphosphohydrolase that can hydrolyze dNTPs into deoxynucleosides and triphosphates to keep the balance of the intracellular dNTPs pool. Moreover, it has been reported that SAMHD1 plays a role in regulating cell proliferation and the cell cycle, maintaining genome stability and inhibiting innate immune responses. SAMHD1 activity is regulated by phosphorylation, oxidation, SUMOylation, and O-GlcNAcylation. SAMHD1 mutations have been reported to cause diseases, including chronic lymphocytic leukemia and mantle cell lymphoma. SAMHD1 expression in acute myeloid leukemia predicts inferior prognosis. Recently, it has been revealed that SAMHD1 mediates the resistance to anti-cancer drugs. This review will focus on SAMHD1 function and regulation, the association between SAMHD1 and hematological malignancies and will provide updated information on SAMHD1 mediating resistance to nucleoside analogue antimetabolites, topoisomerase inhibitors, platinum-derived agents and DNA hypomethylating agents. Furthermore, histone deacetylase inhibitors and tyrosine kinase inhibitors indirectly increase anti-cancer drug resistance by increasing SAMDH1 activity. We herein highlight the importance of the development of novel agents targeting SAMHD1 to overcome treatment resistance of hematological malignancies, which would be an opportunity to improve the outcome of patients with refractory hematological malignancies.

1264

Plasmacytoma variant translocation 1 (PVT1) is a long non-coding RNA (lncRNA) gene identified as a recurrent breakpoint of Burkitt's lymphomas. Human PVT1 gene is located on region 8q24.21, a well-known cancer risk region, and encodes at least 26 linear ncRNA isoforms and 26 circular RNA isoforms, as well as 6 microRNAs. Several PVT1 functioning models have been reported recently such as competing endogenous RNA (ceRNA) activity and regulating protein stability of oncogenes, especially MYC oncogene. The promoter of PVT1 gene is a boundary element of tumor-suppressor DNA. CircPVT1 derived from PVT1 gene is also a critical non-coding oncogenic RNA. Although substantial advancements have been made in understanding the roles of PVT1 in cancer recently, the detailed mechanisms underlying its functions remain unclear. Herein, we summarize the recent progressions on the mechanisms underlying PVT1 regulated gene expression at different levels. We also discuss the interaction between lncRNA and protein, RNA and DNA, as well as the potential cancer therapy strategy by targeting these networks.

1224

Krüppel-like factor 10 (KLF10), also known as TGFb-inducible early gene-1 (TIEG1), was first found in human osteoblasts. Early studies show that KLF10 plays an important role in osteogenic differentiation. Through decades of research, KLF10 has been found to have complex functions in many different cell types, and its expression and function is regulated in multiple ways. As a downstream factor of transforming growth factor b (TGFb) /SMAD signaling, KLF10 is involved in various biological functions, including glucose and lipid metabolism in liver and adipose tissue, the maintenance of mitochondrial structure and function of the skeletal muscle, cell proliferation and apoptosis, and plays roles in multiple disease processes, such as nonalcoholic steatohepatitis (NASH) and tumor. Besides, KLF10 shows gender-dependent difference of regulation and function in many aspects. In this review, the biological functions of KLF10 and its roles in disease states is updated and discussed, which would provide new insights into the functional roles of KLF10 and a clearer view of potential therapeutic strategies by targeting KLF10.

1379

The endometrium is the inner mucosal lining of the uterus that undergoes extensive cyclic growth, regeneration, differentiation, and shedding throughout the menstrual cycle in response to steroid hormones. It repeatedly undergoes approximately 450 cycles of degeneration and regeneration in a woman's lifetime. Endometrial abnormalities can be associated with repeated embryo implantation failure, recurrent spontaneous abortion, and other physiological features responsible for female infertility. This significant regenerative capacity may occur as a result of tissue-resident stem cell populations within the endometrium. Indeed, the existence of endometrial stem cells was only observed in humans and rodents through several isolation and characterization methods in the last few years. Although endometrial stem cells share various biological characteristics with other types of mesenchymal stem cells, they also show some differences in phenotype, self-renewal, and multilineage differentiation potential. Extensive studies over many years on endometrial stem cells will provide new insights into the physiology and mechanisms underlying various gynaecological diseases related to endometrial abnormalities such as female infertility, endometriosis, and endometrial cancer. Here we summarized recent studies about cellular origins and biological characteristics of endometrial stem cells. We also reviewed various recent studies to improve our understanding of their physiological roles. Many preclinical studies on their potential therapeutic applications to various endometrial diseases that could lead to reproductive dysfunction were also reviewed.

1226

Pathological cardiac damage during heart failure is associated with cell death and damage associated molecular patterns (DAMPs) release which triggers a viscous cycle of sterile inflammation to mediate maladaptive cardiac tissue remodelling during the progression to heart failure. DAMPs like cytokines, chemokines, and nuclear or mitochondrial genomic fragments are released in the pathological myocardium. Interestingly, circulating or cytosolic DNA fragments can play a role in the disease by interaction with nucleic acid sensors expressed in cardiomyocyte and non-myocyte neighbouring cells. The circulating cell free DNA (cfDNA) fragments have been clinically reported as markers for various diseases including cardiovascular pathophysiology. Such cfDNA within the DAMP pool can mediate intra-and inter-cellular signalling cascade to upregulate transcriptional expression of inflammatory mediators and trigger oxidative stress within cells. The cellular role of such genomic equivalents varying with chronic or acute stress might be correlated with the cell death forms encountered in myocardium during disease progression. Thus, cfDNA can be phenotypically correlated as a critical player towards upregulation of pathological processes like interstitial fibrosis, cardiomyocyte contractile dysfunction and cell death. Herein, we review the association of cfDNA with heart failure and analyse their potential usage as novel and effective therapeutic targets towards augmentation of cardiac function.

1593

Continuous revision of the histologic and stage-wise classification of lung cancer by the World Health Organization (WHO) provides the foundation for therapeutic advances by promoting molecular targeted and immunotherapies and ensuring accurate diagnosis. Cancer epidemiologic data provide helpful information for cancer prevention, diagnosis, and management, supporting health-care interventions. Global cancer mortality projections from 2016 to 2060 show that cancer will overtake ischemic heart diseases (IHD) as the leading cause of death (18.9 million) immediately after 2030, surpassing non-small cell lung cancer (NSCLC), which accounts for 85 percent of lung cancers. The clinical stage at the diagnosis is the main prognostic factor in NSCLC therapies. Advanced early diagnostic methods are essential as the initial stages of cancer show reduced mortality compared to the advanced stages. Sophisticated approaches to proper histological classification and NSCLC management have improved clinical efficiency. Although immune checkpoint inhibitors (ICIs) and targeted molecular therapies have refined the therapeutic management of late-stage NSCLC, the specificity and sensitivity of cancer biomarkers should be improved by focusing on prospective studies, followed by their use as therapeutic tools. The liquid biopsy candidates such as circulating tumor cells (CTCs), circulating cell-free tumor DNA (cfDNA), tumor educated platelets (TEP), and extracellular vesicles (EVs) possess cancer-derived biomolecules and aid in tracing: driver mutations leading to cancer, acquired resistance caused by various generations of therapeutic agents, refractory disease, prognosis, and surveillance.

1758

1420

Ovarian cancer is the second most fatal gynecological cancer. For the last decade or so significant use of non-circulating and circulating biomarkers has been highlighted. However, the study of such biomarkers at nanovesicle technology such as exosomes, proteomic and genomics studies could further contribute to better identification of anomalous protein and networks which could act as potential targets for biomarker and immunotherapy development. This review provides an overview of the circulating and non-circulating biomarkers with the aim of addressing the current challenges and potential biomarkers that could lead to early ovarian cancer diagnosis and better management. By means of this review we also lay a hypothesis that characterization of exosomal protein, nucleic acid content from body fluids (serum, plasma, urine, etc.) can decode the secret of disease and potentially improve diagnostic sensitivity which could further lead to more effective screening and early detection of the disease.

1286

Little is known about the difference in durability of HBsAg seroclearance induced by nucleoside analogs (NAs) or by interferon (IFN). A real-world, retrospective cohort study was conducted. Patients were assigned into two groups: NAs monotherapy-induced HBsAg seroclearance subjects and IFN monotherapy induced-HBsAg seroclearance subjects. A total of 198 subjects, comprised by 168 NAs monotherapy-induced and 30 IFN monotherapy-induced, who achieved HBsAg seroclearance were included in this study. The one-year probabilities of confirmed HBsAg seroclearance were significantly different in patients with NAs monotherapy and IFN monotherapy (0.960 (with 95% CI 0.922-0.999) vs. 0.691 (with 95% CI 0.523-0.913), log-rank-P = 4.04e-4). 73.3% (11 of 15) HBsAg recurrence occurred within one year after HBsAg seroclearance. The one-year probabilities of confirmed HBsAg seroclearance were higher in IFN monotherapy patients with anti-HBs than in IFN monotherapy patients without anti-HBs (0.839 (with 95% CI 0.657-1.000) vs. 0.489 (with 95% CI 0.251-0.953), log-rank test, P = 0.024). Our study thus provided novel insights into the durability of HBsAg seroclearance induced by NAs or IFN monotherapy. In particular, the HBsAg seroreversion rate was relatively high in IFN monotherapy subjects. The presence of anti-HBs was significantly correlated with a longer durability of functional cure induced by IFN treatment. And one-year follow-up in HBsAg seroclearance achieved individuals is proper for averting HBsAg seroreversion and other liver disease.

1285

Idiopathic pulmonary fibrosis (IPF) is a dreadful, chronic, and irreversibly progressive disease leading to death with a few effective treatments. Our previous study suggested that repetitive hyperbaric oxygen (HBO) treatment alleviates bleomycin-induced pulmonary fibrosis in mice. Here, we investigated the protective mechanism of HBO treatment against pulmonary fibrosis using an integrated approach. Analyzing publicly available expression data from the mouse model of bleomycin-induced pulmonary fibrosis as well as IPF patients, several potential mechanisms of relevance to IPF pathology were identified, including increased epithelial-to-mesenchymal transition (EMT) and glycolysis. High EMT or glycolysis scores in bronchoalveolar lavage were strong independent predictors of mortality in multivariate analysis. These processes were potentially driven by hypoxia and blocked by HBO treatment. Together, these data support HBO treatment as a viable strategy against pulmonary fibrosis.

1331

BMP9 mediated osteogenic differentiation mechanisms of MSCs were widely explored, however, mechanisms of BMP9-induced angiogenesis still need to be clarified. We previously characterized that Notch1 promoted BMP9-induced osteogenesiseangiogenesis coupling process in mesenchymal stem cells (MSCs). Here, we explored the underlying mechanisms of lncRNA H19 (H19) mediated regulation of BMP9-induced angiogenesis through activating Notch1 signaling. We demonstrated that basal expression level of H19 was high in MSCs, and silencing H19 attenuates BMP9-induced osteogenesis and angiogenesis of MSCs both in vitro and in vivo. Meanwhile, we identified that BMP9-induced production of CD31⁺ cells was indispensable for BMP9-induced bone formation, and silencing H19 dramatically blocked BMP9-induced production of CD31⁺cells. In addition, we found that down-regulation of H19 inhibited BMP9 mediated blood vessel formation and followed subsequent bone formation in vivo. Mechanistically, we clarified that H19 promoted p53 phosphorylation by direct interacting and phosphorylating binding, and phosphorylated p53 potentiated Notch1 expression and activation of Notch1 targeting genes by binding on the promoter area of Notch1 gene. These findings suggested that H19 regulated BMP9-induced angiogenesis of MSCs by promoting the p53-Notch1 angiogenic signaling axis.

1423

Small non-coding RNAs are potential diagnostic biomarkers for lung cancer. Mitochondria-derived small RNA (mtRNA) is a novel regulatory small non-coding RNA that only recently has been identified and cataloged. Currently, there are no reports of studies of mtRNA in human lung cancer. Currently, normalization methods are unstable, and they often fail to identify differentially expressed small non-coding RNAs (sncRNAs). In order to identify reliable biomarkers for lung cancer screening, we used a ratio-based method using mtRNAs newly discovered in human peripheral blood mononuclear cells. In the discovery cohort (AUC = 0.981) and independent validation cohort (AUC = 0.916) the prediction model of eight mtRNA ratios distinguished lung cancer patients from controls. The prediction model will provide reliable biomarkers that will allow blood-based screening to become more feasible and will help make lung cancer diagnosis more accurate in clinical practice.

1284

Disrupting the balance of neuronal excitation and inhibition (E/I) is an important pathogenic mechanism of anxiety and depression. Interferon regulatory factor 3 (IRF3) plays a key role in the innate immune response, and activation of IRF3 triggers the expression of type I interferons and downstream interferon-stimulated genes, which are associated with anxiety and depression. However, whether IRF3 participates in the pathogenesis of anxiety/depression by regulating E/I balance remains poorly understood. Here, we reported that global knockout (KO) of IRF3 (IRF3^-/-) significantly increased anxiety/depression-like behaviors, but did not affect normal spatial learning and memory. Compared with wild type (WT) control mice, the E/I balance was disrupted, as reflected by enhanced glutamatergic transmission and decreased GABAergic transmission in the neurons of hippocampal CA1 and medial prefrontal cortex (mPFC) in IRF3-KO mice. Importantly, genetic rescue of IRF3 expression by adeno-associated virus (AAV) was sufficient to alleviate anxiety/depression-like behaviors and restore the neuronal E/I balance in IRF3-KO mice. Taken together, our results indicate that IRF3 is critical in maintaining neuronal E/I balance, thereby playing an essential role in ensuring emotional stability.

1282

Major challenges such as nuclease degradation, rapid renal clearance, non-specific delivery, poor cellular uptake and inflammatory response have limited the clinical application of small RNA-mediated gene silencing. To overcome these challenges, we designed a novel targeting small RNA delivery platform comprising of three oligonucleotides: (1) a guide RNA sequence, (2) part of a passenger sequence linked to a DNA aptamer via a PEG linker, and (3) another passenger sequence conjugated to cholesterol, which assemble through complementary base pair annealing. Remarkably, in the presence of magnesium, this molecule self-assembled into a nanoparticle with a hydrophobic cholesterol core, hydrophilic RNA oligonucleotide shell and PEG-linked DNA aptamer flare. The nanoparticles conferred protection to the RNA oligonucleotides against nuclease degradation, which increased bioavailability, and reduced systemic inflammatory responses. The aptamer allowed targeted delivery of RNA therapeutics through cell-specific surface markers, and once inside the cell, the nanoparticles induced lysosomal leakage that released the RNA oligonucleotides into the cytosol to achieve gene silencing. We created a c-Kit-targeting miR-26a delivery particle that specifically accumulated in c-Kit⁺ breast cancer, significantly increased T cell recruitment, and inhibited tumor growth. Regression of large established tumors were achieved when the nanoparticle was used in combination with anti-CTLA-4 monoclonal antibody.

1252

Nod-like receptor family pyrin domain-containing protein 12 (NLRP12) is one of the critical pattern recognition receptors which participates in the regulation of multiple inflammatory responses. Mutations in NLRP12 cause exceptionally rare NLRP12-associated autoinflammatory disease (NLRP12-AID). So far, very few patients with NLRP12-AID have been identified worldwide; therefore, data on the clinical phenotype and genetic pro file are limited. In this study, we reported 10 patients who presented mainly with periodic fever syndrome or arthritis. Next-generation sequencing (NGS) identified 6 heterozygous mutations of NLRP12, including 2 novel null mutations. Of the patients, some with same mutations showed different clinical features. Compared to healthy controls, the increased levels of cytokines were revealed in the patients'plasmas, as well as in the supernatants of patients'cells stimulated with lipopolysaccharide (LPS) or tumor necrosis factor-a (TNF-a). The missense mutations did not change the protein expression; but decreased level of NLRP12 protein was shown in the null mutations. And in vitro expression assay demonstrated a truncating protein induced by the frameshift mutation. Further functional studies revealed the deleterious effect of mutations on nuclear factor-kappa B (NF-kB) signaling. Both the null and missense mutations impaired their inhibition of NF-kB activation induced by p65. Collectively, this study reported a relatively large NLRP12-AID case series. Our findings expand the clinical spectrum, and reinforce the diversity of genetic mutations and clinical phenotypes. The NLRP12-associated disorder should be considered when autoinflammatory diseases are encountered in the clinical practice, especially for patients presenting with periodic fever but no other genetic cause identified.

1252

Daily insulin injection is necessary for the treatment of the insulin-dependent diabetes. However, the process is painful and inconvenient. Accordingly, we have made exploratory efforts to establish an alternative method for continuous insulin supply via intramuscular injection of a designed plasmid encoding the single-strand insulin analogue (SIA), which provides safe, effective and prolonged control of insulin-dependent diabetes. To generate a SIA, a short flexible peptide was alternatively introduced into the natural proinsulin to replace its original long and rigid C-peptide. Then, the synthetic promoter SP301 was used to drive potent and specific expression of SIA in skeletal muscle cells. By combining the Pluronic L64 and low-voltage electropulse (L/E), the specialized gene delivery technique was applied to efficiently transfer the constructed plasmid into skeletal muscle cells via intramuscular injection. Through these efforts, a plasmid-based intramuscular gene expression system was established and improved, making it applicable for gene therapy. The plasmidexpressed SIA showed biological functions that were similar to that of natural insulin. A single L/E-pSP301-SIA administration provided sustained SIA expression in vivo for about 1.5 months. In addition, the diabetic mice treated with L/E-pSP301-SIA were much healthier than those with other treatments. This plasmid-based system was safe for the treatment of diabetes and did not cause immune responses or pathological damage. The results confirmed that, in a mouse model, long-term positive effects were achieved by a single intramuscular L/E-pSP301-SIA injection, which consequently provided reliable experimental basis for its clinical application for the treatment of diabetes mellitus with promising prospects.

1863

Macrophages (Mφs) play a crucial role in the pathological progression of osteoarthritis (OA) by regulating inflammation and tissue repair. Decreasing pro-inflammatory M1-Mφs and increasing anti-inflammatory M2-Mφs can alleviate OA-related inflammation and promote cartilage repair. Apoptosis is a natural process associated with tissue repair. A large number of apoptotic bodies (ABs), a type of extracellular vesicle, are produced during apoptosis, and this is associated with a reduction in inflammation. However, the functions of apoptotic bodies remain largely unknown. In this study, we investigated the role of M2-Mφs-derived apoptotic bodies (M2-ABs) in regulating the M1/M2 balance of macrophages in a mouse model of OA. Our data show that M2-ABs can be targeted for uptake by M1-Mφs, and this reprograms M1-to-M2 phenotypes within 24 h. The M2-ABs significantly ameliorated the severity of OA, alleviated the M1-mediated pro-inflammatory environment, and inhibited chondrocyte apoptosis in mice. RNA-seq revealed that M2-ABs were enriched with miR-21-5p, a microRNA that is negatively correlated with articular cartilage degeneration. Inhibiting the function of miR-21-5p in M1-Mφs significantly reduced M2-ABs-guided M1-to-M2 reprogramming following in vitro cell transfection. Together, these results suggest that M2-derived apoptotic bodies can prevent articular cartilage damage and improve gait abnormalities in OA mice by reversing the inflammatory response caused by M1 macrophages. The mechanism underlying these findings may be related to miR-21-5p-regulated inhibition of inflammatory factors. The application of M2-ABs may represent a novel cell therapy, and could provide a valuable strategy for the treatment of OA and/or chronic inflammation.

1351