Autophagy, the process for recycling cytoplasm in the lysosome, relies on tightly regulated membrane trafficking. During autophagy, autophagosomes either fuse with endosomes generating amphisomes and then lysosomes, or directly fuse with lysosomes, in both cases generating autolysosomes that degrade their contents. It remains unclear whether specific mechanisms or conditions determine these alternate routes. Here, we demonstrate that the endosomal regulator SNX3 specifically regulates basal autophagy under nutrient-adequate conditions in both Caenorhabditis elegans (C. elegans) and cultured mammalian cells. In C. elegans, SNX-3 depletion elevates autophagy independently of the UNC-51/ULK1 complex and leads to the accumulation of both autophagosomes and amphisomes, which consequently impairs the clearance of autophagic cargo, including SQST-1/p62 and protein aggregates. Mechanistically, SNX-3 depletion differentially regulates the machineries required for autophagosome-lysosome fusion. In snx-3 mutants, the Q-SNARE components SYX-17 and SNAP-29 translocate to autophagosomes, where they assemble with the endosomal R-SNAREs VAMP-7 and VAMP-8 to promote amphisome formation. Conversely, loss of SNX-3 impairs the lysosomal delivery of VAMP-8 and RAB-7, both essential for autophagosome/amphisome-lysosome fusion, thereby generating fusion-incompetent lysosomes. However, starvation restores the lysosomal fusion capability compromised by snx-3 depletion. Our findings reveal that autophagosome-lysosome fusion is preferentially regulated by nutrient status, and identify an endosomal regulator that tunes membrane trafficking with changing autophagy demands.

Natural products represent a vital source for drug discovery targeting G protein-coupled receptors (GPCRs). Among them, bioactive peptides have emerged as promising therapeutic candidates owing to their high selectivity and low toxicity. In this study, we investigated the modulatory effects of peptides derived from Saccharomyces cerevisiae on the angiotensin II (AngII) type 1 receptor (AT1R), a class A GPCR that plays a central role in cardiovascular homeostasis. We identified a heptapeptide TLPPPPL (TLP) that enhances AngII binding to AT1R without altering maximal binding capacity, as determined by an HTRF-based ligand-binding assay. Functional analyses using BRET-based biosensors revealed that TLP acts as a positive allosteric modulator, potentiating both G protein activation and β-arrestin recruitment. Furthermore, TLP augments signaling induced by biased AT1R agonists and rescues the impaired function of disease-associated AT1R mutants, indicating a broad positive modulatory effect on agonist-driven receptor activation. Together, these findings identify a yeast-derived positive allosteric modulator of AT1R and underscore the potential of natural peptides as promising leads for the development of GPCR-targeted therapeutics.

Background Because gain of chromosome 3q occurs early in cervical carcinogenesis, amplification of the telomerase RNA component gene (TERC), which is located at 3q26, represents a promising biomarker for the diagnosis and prognosis of cervical neoplasia. However, current screening approaches face limitations, including low sensitivity and high variability,which restrict their clinical utility. Furthermore, the optimal method for interpreting TERC fluorescence in situ hybridization (FISH) remains unclear. Methods To address this issue, we carried out both retrospective and prospective analyses. First, we assessed TERC gene expression and its prognostic value using the cBioPortal database and Kaplan-Meier survival analysis. We further selected 200 biopsy specimens with available valid Pap test records.Additionally, we performed human papillomavirus (HPV) genotyping and hematoxylin Eosin (HE) stain on serial cervical formalin-fixed paraffin-embedded (FFPE). Finally, we detected TERC gene amplification, and telomerase activity using quantitative (q)PCR and FISH. Results In this study, cytology and histology tests showed significantly different detection rates (P < 0.0001). High-risk HPV was detected in 149 cases (84.2%), with 28 cases (15.8%) testing negative for all 14 high-risk subtypes. Notably, infections with HPV 16, 18 and 52 were significantly more frequent compared with other subtypes and infection with multiple HPV subtypes was statistically significant (P < 0.0001). TERC gene amplification was detected by FISH in 93.8% of all biopsies (P < 0.0001). Consistently, qPCR yielded similar results. The TERC amplification rate was significantly higher in cervical intraepithelial neoplasia and squamous cell carcinoma (P = 0.0001). Additionally, the number of TERC-positive signals was positively correlated with the severity of cervical lesions, which aligns with the acquisition of extra 3q copies (P < 0.05). TERC-FISH exhibited a good diagnostic performance. Conclusions The present findings reaffirm and support the potential of TERC as a biomarker for both the diagnosis and prognosis of cervical neoplasia. This study recommends that TERC-FISH can be used with caution to confirm patient status in cases with discrepancies in the histo-cytology diagnosis.

Erythropoiesis requires precise coordination of transcriptional and co-/post-transcriptional programs, yet the role of alternative polyadenylation (APA) in this process remains poorly understood. Here, we profiled the genome-wide dynamic APA landscape during erythropoiesis using single-cell RNA sequencing (scRNA-seq). Through clustering and functional enrichment analysis, seven distinct APA dynamic patterns were identified, with genes showing stage-specific APA changes enriched in erythroid lineage differentiation, heme synthesis, and iron metabolism. Combining motif analysis near polyadenylation sites (PASs) and APA regulators expression profiling, we observed that cleavage and polyadenylation specificity factor 6 (CPSF6), a critical APA regulator, exhibited significant variation. Functional assays demonstrated that CPSF6 facilitates erythropoiesis, as its depletion impaired heme synthesis and intracellular iron deficiency. Mechanistically, CPSF6 depletion shortened the 3 ' UTR length of iron metabolism regulators (FAM210B, IREB2, TFRC), which was accompanied by reduced expression of these genes. Clinically, CPSF6 and these APA-regulated iron metabolism-related genes were aberrantly upregulated in polycythemia vera (PV) patients, correlating with erythroid hyperproliferation. Collectively, our findings support a CPSF6-APA-iron homeostasis axis as an important co-/post- transcriptional regulatory mechanism in erythropoiesis, and implicate its dysregulation in the pathogenesis of PV, offering novel molecular targets for therapeutic intervention in myeloproliferative neoplasms.

Ovarian cancer remains a leading cause of gynecologic cancer mortality, largely due to high recurrence and frequent cisplatin resistance. This study investigates the role of ADAR1 lactylation-mediated RNA editing in ovarian cancer chemoresistance. Cisplatin-resistant cells exhibit significantly elevated global adenosine-to-inosine (A-to-I) RNA editing and ADAR1 expression. Genetic knockdown of ADAR1 enhances cisplatin sensitivity in vitro and in vivo, activates innate immune MAVS/PKR pathways, and promotes CD4⁺/CD8⁺ T cell infiltration. Mechanistically, Tip60 mediates ADAR1 lactylation, facilitating its interaction with deubiquitinase USP48 to stabilize ADAR1. Lactylation-dependent ADAR1 upregulation suppresses innate immunity and enhances protein translation, driving chemoresistance. Notably, inhibitor ZYS-1 targets ADAR1 lactylation, reducing ADAR1 expression, activating antitumor immunity, and synergizing with cisplatin to delay tumor growth in mice. These findings establish ADAR1 lactylation as a critical regulatory mechanism, supporting ZYS-1 plus cisplatin as a promising strategy for platinum-resistant ovarian cancer.

Mesenchymal stem cells (MSCs) have attracted considerable attention for clinical translation in regenerative medicine, primarily due to their validated paracrine effects, prominent immunomodulatory properties, and superior multipotent differentiation capabilities. However, the limited homing efficiency and poor post-transplant survival of MSCs severely compromise therapeutic efficacy, thereby giving rise to suboptimal and inconsistent treatment outcomes. To circumvent these critical drawbacks and fully harness the therapeutic potential of MSCs, researchers have incorporated a diverse array of effective strategies, including genetic engineering, preconditioning with cytokines, small molecular compounds or hypoxic stimuli, and scaffold-based culture systems. Given these promising research advances, this review systematically summarizes recent advances in MSC-enhanced therapeutic strategies, and elaborates on their core molecular mechanisms as well as how these mechanisms modulate MSC survival, homing capacity and immunomodulatory efficacy. On this basis, we further analyze the practical applicability of these enhanced MSCs in clinical trials and seek to provide critical insights for the clinical selection of MSC-based enhanced therapies.

Abstract Background Pseudogene‐derived lncRNAs are widely dysregulated in cancer. Technological advancements have facilitated the functional characterization of increasing pseudogenes in cancer progression. However, the association between pseudogenes and RNA N6‐methyladenosine (m6A) modification in cancer, as well as the underlying mechanisms, remains largely unexplored. Methods We analyzed the expression of 12 146 pseudogenes and comprehensively examined the m6A modification of RNAs derived from them and their paralogs. Through integrative analysis of multi‐omics data, we explored the associations between pseudogene dysregulation and m6A, identifying critical pseudogenes involved in HGSOC progression. Tumour promotion role of RPS15AP12 and its cognate parent gene was characterized by cell proliferation, transwell assays, and scratch assays in ovarian cells and xenograft nude mice. RNA decay assays were used to reveal the participation of m6A in decreasement of RPS15AP12 lncRNA stability. Luciferase reporter assays were performed to verify that RPS15AP12 enhances RPS15A expression by competitively binding to miR‐96‐3p. Western blot and phosphorylation assays were performed to investigate the impairment of RPS15AP12 towards the sensors of MAVS (RIG‐I and MDA5), and downstream p‐TBK1 and p‐IRF3. Finally, ELISA assays were performed to explore the regulatory role of RPS15AP12 in IFN‐β expression. Results M6A is distributed across over a thousand pseudogenes, and hypomethylation leads to their upregulation in HGSOC. We identified a processed pseudogene, RPS15AP12, upregulated by FTO‐mediated m6A demethylation. RPS15AP12 enhances the growth ability and metastatic capabilities of ovarian cancer (OC) cells via functioning as a competitive endogenous RNA (ceRNA) for its host gene, RPS15A, through the sequestration of miR‐96‐3p. Importantly, the deletion of RPS15AP12 diminishes the expression of RPS15A, leading to the upregulation of anti‐tumour immune responses by activating RIG‐I and MDA5 and downstream p‐TBK1 and p‐IRF3 as well as IFN‐β levels. Conclusion Our findings expand the understanding of m6A‐modulated pseudogenes in tumour growth and anti‐tumour innate immunity in OC. Key Points Genome‐wide profiling reveals the redistribution of m6A modification on pseudogene‐derived lncRNAs and m6A redistribution‐relevant dysregulation of pseudogenes in HGSOC. RPS15AP12, as a representative processed pseudogene, is up‐regulated by FTO‐mediated demethylation and acts as a miRNA sponge to promote RPS15A expression via competitively binding to miR‐96‐3p. RPS15AP12/RPS15A axis inhibits MAVS sensors (RIG‐I and MDA5) and downstream IFN‐β levels in ovarian cancer.

Rescued effect of NUMB-L overexpression on the cell proliferation and metastasis in SRSF9-depleted ovarian cancer cells.

To investigate the impact of COVID-19 infection on maternal and neonatal outcomes and immunity in pregnant women in China. 283 pregnant women with COVID-19 were included in the prospective observational cohort study and divided into five groups based on infection stage. Antibody levels were measured in plasma, umbilical cord blood, and breast milk, and combined with clinical data and 6-month follow-up results. We measured SARS-CoV-2 antibody levels using a chemiluminescence immunoassay and analyzed the data with the Kruskal-Wallis test, χ2 test, or Fisher’s exact test. No significant differences were found in age, BMI, weight change during pregnancy, or the incidence of gestational hypertension, gestational diabetes, gestational hypothyroidism, intrahepatic cholestasis, transaminitis, preterm birth, small for gestational age, neonatal NICU transfers, developmental delays, and hearing damage among the five groups. The incidence of COVID-19 in infants from mothers infected at different stages of pregnancy was significantly lower than in the uninfected group (P < 0.05). Maternal and umbilical cord blood showed significantly higher IgG levels in the infected group compared to the uninfected group at different stages of pregnancy (P < 0.05). The median transplacental antibody transfer ratio across all infection groups was 1.15 (0.98–1.30), with no significant differences between them. The reinfection group had significantly higher IgA levels during pregnancy compared to other groups (P < 0.05). No adverse outcomes were observed in mothers or infants at any stage of maternal SARS-CoV-2 infection. Antibodies in umbilical cord blood and breast milk may offer passive immunity to newborns for 1–3 months. Reinfection during pregnancy may extend this immunity without raising the risk of adverse outcomes.

Abnormal metabolism is a typical characteristic of malignant tumors, broadly influencing the outcome of cancer therapy. Research on tumor metabolism has increased exponentially in last decades. There is an urgent need to provide a blueprint or framework to accelerate the translation of cancer metabolism research into effective therapeutics. In this review, we summarize the impact of metabolism on cancer progression and emerging metabolic therapies, and systematically outline potential clinical applications in cancer prevention, early screening, diagnosis, treatment, and new challenges. Furthermore, we aim to provide researchers and clinicians with a clearer perspective on the immediate translation of clinical applications stemming from cancer metabolism research. It will revolutionize cancer management and significantly enhance patient survival.

Ovarian cancer (OC) is one of the deadliest gynecological malignancies. As the prevalent post-transcriptional regulation, alternative polyadenylation (APA) plays a crucial role in various tumors. Here we identify that the APA regulator NUDT21 is upregulated in OC and promotes malignant progression. We further demonstrate that IGF2BP3 interacts with NUDT21, which suggests m6A modification could regulate APA processing. Mechanistically, IGF2BP3, recognizing the m6A-modified site in intron 32 of SPTBN1, recruits NUDT21 to promote the usage of the SPTBN1 proximal polyadenylation site (PAS), thus increasing the generation of short transcripts in OC cells. Intriguingly, the SPTBN1 long variant demonstrates tumor-suppressive properties, whereas the short variant enhances oncogenic activity in OC. Subsequently, we illustrate that the long isoform inhibits tumor growth and metastasis by binding to CDK1 and blocking the G2/M phase of the cell cycle. In conclusion, this study uncovers a previously unrecognized regulatory mechanism in OC, which could provide potential therapeutic strategies for OC.

P4-ATPases are phospholipid flippases responsible for the transbilayer lipid asymmetry. ATP8A1, a P4-ATPase family member, has been reported to be involved in phosphatidylserine (PS) translocation at the trans-Golgi network, early endosomes and recycling endosomes. However, the possible roles of the PS on late endosomes/lysosomes pathway and how they are regulated remain to be elucidated. This study showed enrichment of ATP8A1 in Rab7-positive late endosomal compartments, and that ATP8A1 primarily flips the endosomal PS from the luminal leaflet to the cytosolic leaflet but not the PS in the inner leaflet of the plasma membrane. ATP8A1 depletion accelerates the lysosome-destined cargo proteins transfer into the intraluminal vesicles (ILVs) of multivesicular bodies (MVBs) and alters the signaling of epidermal growth factor receptor. Mechanistically, ATP8A1 depletion leads to PS loading in the luminal leaflet of MVB's limiting membrane, which fine-tunes ILVs initiation and endosomal sorting complex required for transport (ESCRT) component recruitment.

Neurodegenerative diseases (NDDs), including Alzheimer’s disease (AD), Parkinson’s disease (PD), amyotrophic lateral sclerosis (ALS), Huntington’s disease (HD), and prion disease, represent a group of age-related disorders that pose a growing and formidable challenge to global health. Despite decades of extensive research that has uncovered key genetic factors and biochemical pathways, the precise molecular mechanisms underlying these diseases and effective therapeutic strategies remain elusive. Caenorhabditis elegans (C. elegans) has emerged as a powerful model organism for studying NDDs due to its unique biological features such as genetic tractability, conserved molecular pathways, and ease of high-throughput screening. This model provides an exceptional platform for identifying molecular targets associated with NDDs and developing novel therapeutic interventions. This review highlights the critical role of C. elegans in elucidating the complex molecular mechanisms of human NDDs, with a particular focus on recent advancements and its indispensable contributions to the discovery of molecular targets and therapeutic strategies for these NDDs.

Ovarian cancer has the highest mortality rate among gynecologic tumors worldwide, with unclear underlying mechanisms of pathogenesis. RNA-binding proteins (RBPs) primarily direct post-transcriptional regulation through modulating RNA metabolism. Recent evidence demonstrates that RBPs are also implicated in transcriptional control. However, the role and mechanism of RBP-mediated transcriptional regulation in tumorigenesis remain largely unexplored. Here, we show that the RBP heterogeneous ribonucleoprotein L (hnRNPL) interacts with chromatin and regulates gene transcription by forming phase-separated condensates in ovarian cancer. hnRNPL phase separation activates PIK3CB transcription and glycolysis, thus promoting ovarian cancer progression. Notably, we observe that the PIK3CB promoter is transcribed to produce a non-coding RNA which interacts with hnRNPL and promotes hnRNPL condensation. Furthermore, hnRNPL is significantly amplified in ovarian cancer, and its high expression predicts poor prognosis for ovarian cancer patients. By using cell-derived xenograft and patient-derived organoid models, we show that hnRNPL knockdown suppresses ovarian tumorigenesis. Together, our study reveals that phase separation of the chromatin-associated RBP hnRNPL promotes PIK3CB transcription and glycolysis to facilitate tumorigenesis in ovarian cancer. The formed hnRNPL-PIK3CB-AKT axis depending on phase separation can serve as a potential therapeutic target for ovarian cancer.

Sequences of shRNA oligos and antisense oligonucleotides used in the study.