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Li, N.

Publications and source records attributed to Li, N..

At least 73 records · Page 4Linked to original sources

Tissue-Specific and NF-kappaB-Independent Xist RNA Localization Patterns in Female Intestinal, Blood, and Muscle Progenitors

X-Chromosome Inactivation (XCI), driven by the expression of Xist RNA and the enrichment of various repressive epigenetic marks, results in the formation of a mostly inactive X chromosome (Xi) to equalize X-linked gene expression between sexes. Unexpectedly, in unstimulated female lymphocytes that are largely quiescent, these epigenetic features are largely absent but are restored by NF-{kappa}B signaling following their activation. To determine whether these epigenetic phenotypes correlate with quiescence or NF-{kappa}B activation in other tissues, we evaluated female progenitor and stem cells from intestine, blood, and muscle. Despite known NF-{kappa}B activation, intestinal progenitors have variable Xist RNA patterns, whereas blood progenitors and neutrophils show a strong correlation between NF-{kappa}B activation status and Xist RNA localization. In contrast, muscle satellite cells (SCs) and myoblasts exhibit Xist RNA accumulation at the Xi without NF-{kappa}B activation. Xist RNA localization patterns in SCs change with age, yet adult SCs have an Xi that is mostly transcriptionally silent while allowing expression of muscle-specific X-linked genes including Dmd. These findings reveal that female somatic cells employ diverse, tissue-specific epigenetic mechanisms to maintain X chromosome inactivation, enabling cell type specific Xi gene expression while preserving chromosome-wide silencing.

genetics↗

The neuro-ocular costs of texting during driving

Texting while driving is among the most dangerous forms of distraction, yet the neurophysiological mechanisms linking cognitive load, gaze control, and driving behavior remain poorly understood. Here, EEG, eye tracking, head kinematics, and driving performance were recorded in a naturalistic driving task while participants solved arithmetic problems presented on a dashboard-mounted display under low and high working-memory (WM) load, mimicking real-world interaction with dashboard media. High WM load resulted in slower reaction times, reduced accuracy, prolonged gaze toward the stimulus, increased corrective head rotations, and compensatory reductions in driving speed. Time-frequency analysis revealed robust alpha power suppression ([~]8-13 Hz) over occipito-parietal regions scaling with WM load and closely paralleling gaze behavior. Source reconstruction further identified a transient, stimulus-locked increase in oscillatory power at [~]15 Hz involving the ipsilateral cerebellum and posterior parietal preceding the head rotation onset. These findings illuminate the time course of a neuro-ocular action circuit involving cerebellum and cortex as candidate neurophysiological precursor of upcoming behavioral costs as a result of driver distraction. Bridging cognitive neuroscience and applied traffic research, the present results provide an ecologically valid framework for studying action-attention coupling in real-world behavior.

neuroscience↗

Xenopus IgX informs engineering strategies of IgM and IgG hexamers

Polymeric immunoglobulins are essential components of the immune system in jawed vertebrates. While mammalian immunoglobulin M (IgM) typically forms a pentamer linked by the joining chain (J-chain), Xenopus laevis IgX assembles into a J-chain-independent polymer. Here, we present the cryo-electron microscopy (cryo-EM) structure of IgX, revealing its hexameric configuration. By incorporating the IgX tailpiece into human IgM, we achieved efficient IgM hexamer formation. Truncating IgMs natural tailpiece to a range of 11-16 residues also substantially enhanced hexamerization efficiency. Furthermore, introducing a shortened IgM tailpiece to IgG resulted in effective IgG hexamer formation. We further show that the engineered IgM and IgG hexamers targeting CD20 demonstrated robust complement-dependent cytotoxicity (CDC) against several B lymphoma cells. Additionally, the IgG-Fc hexamer functioned as a decoy, attenuating CDC in cell cultures. These findings deepen our understanding of polymeric immunoglobulin evolution and introduce innovative strategies for the development of IgM- and IgG-based biologics. One Sentence SummaryThe cryo-EM structure of the African clawed frog immunoglobulin IgX reveals a uniform hexameric assembly, inspiring innovative strategies for the development of therapeutic human IgM and IgG hexamers.

immunology↗

A Paracrine-to-Autocrine Shunt of GREM1 Fuels Colorectal Cancer Metastasis via ACVR1C

Tumor cells typically rely on paracrine stromal signals to guide malignant behaviors, but whether they acquire signaling autonomy to support metastasis remains unclear. We elucidate this in colorectal cancer (CRC) by uncovering a paracrine-to-autocrine shunt of Gremlin1 (GREM1), a canonical stromal-secreted antagonist of bone morphogenetic proteins (BMPs). We demonstrate that while GREM1 remains restricted to stromal cells in earlier-stage (I-III) CRC, its ectopic expression in tumor epithelium increases markedly in stage IV. Mechanistically, we identify ACVR1C as a novel, high-affinity epithelial receptor for GREM1. Their interaction activates SMAD2/3 signaling, which upregulates SNAI1 and GREM1, establishing a feedback loop that amplifies epithelial-mesenchymal transition (EMT). Disrupting this loop impairs CRC metastasis in vivo. Clinically, epithelial GREM1 or ACVR1C expression predicts metastasis and poor survival. These findings define a paradigm in which tumor cells hijack stromal GREM1 to establish a GREM1-ACVR1C autocrine loop that sustains EMT and metastasis, marking a shift toward signaling autonomy and revealing a targetable vulnerability in advanced CRC.

cancer biology↗

Tryptophanol, a novel auxin analog found in marine diatoms, enhances nitrogen assimilation

Diatoms exhibit superior competitive capacity in nitrogen assimilation, largely contributing to their growth, although the mechanisms underpinning their success have not been completely understood. Here, a non-ribosomal peptide synthase-like (PtNRPS1, with an unusual domain structure A-T-R1-R2) gene was found to play a vital role in short-term nitrogen assimilation in the marine diatom Phaeodactylum tricornutum. In vitro biochemical assays and in vivo overexpression confirmed that PtNRPS1 catalyzed two sequential two-electron reductions of L-tryptophan to tryptophanol. Tryptophanol exhibits high structural and functional similarities to indole-3-acetic acid (IAA), the most typical phytohormone auxin. Surprisingly, the effective concentration of tryptophanol was lower than that of IAA by as much as 2-5 orders of magnitude for P. tricornutum. Compared with the action of IAA, a distinct molecular mode for tryptophanol was revealed by transcriptomic analysis, resulting mainly in enhanced short-term nitrogen assimilation, which was also confirmed by the elevated nitrogen uptake rates determined by stable-isotope tracking. Finally, global distribution of PtNRPS1 homologues from stramenopiles was found to be positively correlated with the abundance of genes involved in nitrogen assimilation pathways. Overall, our study provides evidence of an auxin-like derivative synthesized by an NRPS in a diatom. We speculate that tryptophanol may accelerate nitrogen assimilation, conferring advantages in the competition for nitrogen in the ocean.

plant biology↗

Cryo-EM Structure of the Erlin1/2 Complex Reveals a Dynamic Scaffold for Membrane Protein Sequestration and Microdomain Assembly

The SPFH (Stomatin, Prohibitin, Flotillin, and HflK/C) family of proteins are key scaffolding components involved in the organization of functional membrane microdomains (FMMs) across various subcellular membranes, including those of the endoplasmic reticulum (ER) and mitochondria, which are characterized by a low content of saturated lipids. Among this protein family, the erlin1/2 complex is specifically located on the ER membrane. Previous studies have shown that the erlin1/2 complex plays an essential role in the ER-associated degradation (ERAD) pathway, mediating the ubiquitin-dependent degradation of various proteins such as inositol 1,4,5-trisphosphate receptors (InsP3Rs), which are important calcium ion transporters on the ER membrane. However, the molecular mechanisms underlying erlin-mediated FMMs organization and its role in ERAD remain poorly understood. In this study, we determined the single-particle cryo-electron microscopy (cryo-EM) structure of the erlin1/2 complex under different detergent conditions. Our findings reveal that the erlin1/2 complex forms a 26-mer cage-like structure, composed of alternating erlin1 and erlin2 subunits. The erlin1/2 complex defines a nanodomain on the ER membrane, which could recruit various types of proteins to both the interior and exterior membrane regions of the cage. By caging cargo proteins, the erlin1/2 complex physically secludes them from their binding partners, leading to a potential halt in their function. Moreover, individual cages can further interact with one another, facilitating the organization of FMMs of different sizes on the ER membrane. These dynamic properties may play a general and critical role in various processes occurring on the ER, including viral replication, positioning the erlin1/2 complex as a promising new target for antiviral drug development.

biophysics↗

The XPF-like domain in SHOC1 required for homologous recombination and safeguarding autosome from meiotic silencing of unsynapsed chromatin

During meiosis, a group of evolutionarily conserved ZMM proteins plays essential roles in stabilizing the recombination intermediates and promoting crossover (CO) formation. In mice, SHOC1 forms a trimeric complex with the other two ZMM proteins, SPO16 and TEX11, to bind recombination intermediates after strand invasion. Although genetic variants of SHOC1 are clinically associated with meiotic arrest and male infertility, their precise molecular mechanisms and evolutionarily conserved functions in human gametogenesis remain enigmatic. Here, we delineated species-specific divergences between human and mouse SHOC1 complex, and identified a missense variant within the XPF-like domain in SHOC1 (c.A1769G:p.Q590R) that was associated with meiotic arrest and non-obstructive azoospermia (NOA). The disorder of the XPF-like domain in SHOC1 impaired DNA double-strand breaks repair by compromising its ability to bind branched DNA structures and the recruitment of M1AP, REDIC1, and ZMM factors to recombination intermediates, ultimately abolishing CO formation. Furthermore, the variant disrupted dynamic 3D chromatin structure in pachytene spermatocytes and induced defects in homologous chromosome synapsis. More importantly, the XPF-like domain in SHOC1 was revealed to prevent autosome intrusion into the sex body compartment, thereby safeguarding critical autosomal loci from meiotic silencing of unsynapsed chromatin (MSUC). Overall, our study demonstrated that the XPF-like domain in SHOC1 is required for homologous recombination and safeguarding autosome from MSUC in meiosis.

developmental biology↗

Testicular mRNA-LNP Delivery: A Novel Therapy for Genetic Spermatogenic Disorders

Uniform testicular maturation arrest is a severe form of male infertility characterized by the presence of germ cells that do not complete spermatogenic development. It is usually caused by meiotic arrest with genetic variants and difficult to treat via drugs or surgery. mRNA-lipid nanoparticle (LNP) delivery is a promising therapeutic option for maturation arrest with monogenic variants via protein replacement therapy. Herein, a spermatocytes-tropic LNP (Pool1-LNP3) was identified via a library of 30 ionizable lipids screening. And in vivo delivery of this novel LNP composition using rete testis microinjection was showed to be high spermatocytes targeting with high transfection efficiency. Thereafter, it was revealed that in vivo delivery of Pool1-LNP3 encapsulating Msh5 mRNA could promote crossover formation and restore spermatogenesis in Msh5D486Y/D486Y mouse models with DSB recombination defects. Notably, the offspring without genomic integration was born using intracytoplasmic sperm injection (ICSI) derived from rescue of Msh5D486Y/D486Ymouse and embryo transfer. Furthermore, no obvious inflammation and histologic damage in any tissue were detected after in vivo delivery of mRNA-LNP. In addition, it was demonstrated that Maps mRNA-LNP3 recovered spermatogenesis in Maps KO mouse with meiotic arrest. Altogether, these findings suggested that this spermatocytes-tropic mRNA-LNP delivery could become a viable and broad applicable strategy for treatment of spermatogenic disorders with genetic defects, providing a foundation for future clinical application.

molecular biology↗

A novel role for UPF1 in safeguarding DNA replication in S phase and beyond.

Accurate DNA replication is essential for the faithful transmission of genetic information to daughter cells. Disruption of this process leads to replication stress, which can trigger mutagenesis, double-stranded DNA breaks, and genomic instability. Transcription is a well-established source of replication stress, contributing through altered chromatin dynamics, DNA structural changes, and direct transcription-replication collisions (TRCs). Here, we uncover a novel role for the RNA/DNA helicase UPF1 in maintaining replication fidelity and responding to replication stress. We show that cancer cells deficient in UPF1 exhibit elevated levels of spontaneous, transcription-dependent replication fork stalling and double-stranded breaks, along with heightened sensitivity to Rad51 and PARP1 inhibitors. Paradoxically, these cells also display resistance to exogenous replication stress, with reduced replication fork stalling, diminished mitotic delays, and decreased activation of mitotic DNA synthesis (MiDAS), a key salvage pathway under stress conditions. Low UPF1 expression has been previously linked to drug resistance in renal carcinoma and correlates with poor prognosis across multiple cancer types. Our findings position UPF1 as a critical guardian against transcription-associated replication stress and suggest that UPF1 deficiency may underlie mechanisms of cancer therapy resistance whilst leaving cells vulnerable to the depletion of key DNA repair pathways. Targeting this vulnerability could offer a promising avenue for therapeutic intervention in UPF1-low tumours.

cell biology↗

An autonomous AI agent for universal behavior analysis

Behavior analysis across species represents a fundamental challenge in neuroscience, psychology, and ethology, typically requiring extensive expert knowledge and labor-intensive processes that limit research scalability and accessibility. We introduce BehaveAgent, an autonomous multimodal AI agent designed to automate behavior analysis from video input without retraining or manual intervention. Unlike conventional methods that require manual behavior annotation, video segmentation, task-specific model training, BehaveAgent leverages the reasoning capabilities of multimodal large language models (LLM) to generalize across novel behavioral domains without need for additional training. It integrates LLMs, vision-language models (VLMs), and large-scale visual grounding modules, orchestrated through a multimodal context memory and goal-directed attention mechanism, to enable robust zero-shot visual reasoning across species and experimental paradigms, including plants, insects, rodents, primates, and humans. Upon receiving a video input, BehaveAgent autonomously identifies the correct analysis strategy and performs end-to-end behavior analysis and interpretation without human supervision. Leveraging vision-language representations, it performs general-purpose tracking, pose estimation and segmentation. We demonstrate BehaveAgents universal applicability to autonomously (1) identify the behavioral paradigm and develop an action plan specialized for the identified paradigm, (2) identify relevant subjects and objects, (3) track those features, (4) identify behavioral sequences with explicit reasoning, (5) generate and execute code for targeted analysis and (6) generate comprehensive research reports that integrate behavioral findings with relevant scientific literature. Through interpretable agentic reasoning, BehaveAgent makes its internal decision-making process transparent, clarifying why particular features are tracked or behaviors inferred. By reducing the time and expertise required for behavior analysis, BehaveAgent introduces a scalable, generalizable, and explainable paradigm for advancing biological and behavioral research.

neuroscience↗

Single-Cell RNA Sequencing and Inferred Protein Activity Analysis Reveal a Distinct Tumor Phenotype in Early-Onset Colorectal Cancer Patients

Colorectal cancer (CRC) diagnosed before age 50 years (early-onset CRC, EO-CRC) is rising at an alarming rate, yet its molecular and microenvironmental drivers remain poorly understood. EO-CRC is highly heterogeneous, and while subtle differences from late-onset CRC (LO-CRC) have been reported, their full extent remains unresolved due to the limited scope of previous studies. Here, we integrate public data with in-house clinical samples profiled by single-cell RNA sequencing (scRNA-seq) and multiplex immunofluorescence (mIF) to compare EO-CRC and LO-CRC. Additionally, we employ gene regulatory network-based protein activity inference (VIPER), enabling a more precise characterization of key regulatory proteins driving tumor-stroma interactions. Our analysis reveals that EO-CRC and LO-CRC have a largely similar immune composition, challenging previous reports of an "immune-cold" phenotype in EO-CRC. However, we identify distinct stromal differences, including a significant enrichment of fibroblasts in EO-CRC. Notably, we define a previously unrecognized epithelial subpopulation in EO-CRC, marked by high expression of toll-like receptor 4 (TLR4) and C-C chemokine receptor type 5 (CCR5)--key mediators of inflammation-driven tumor progression and fibroblast recruitment. These findings suggest that EO-CRC may be driven by a tumor-intrinsic inflammatory phenotype with enhanced stromagenesis, providing new insights into potential mechanisms underlying its increasing incidence in young adults.

cancer biology↗

The spliceosome component U1-70k loads nascent transcripts onto FVE/SGS3 via co-condensation to embark on RNA silencing

Despite biochemical framework of RNA silencing has been studied well, how nascent transcripts enter RNA silencing remains poorly understood. Here we recovered U1-70k, a canonical spliceosome factor, as a novel component in posttranscriptional gene silencing (PTGS). We show that U1-70k interacts with SGS3 and FVE, two key components that supply RNA substrate for RNA-dependent RNA polymerase 6 (RDR6) to generate dsRNA. U1-70k promotes the loading of nascent RNA into the SGS3/FVE complex in the nucleus, which is subsequently translocated onto RDR6 in the cytosol. Importantly, U1-70k contains C-terminal low complexity (LC) domain and displays liquid-liquid phase separation (LLPS) pattern. U1-70k forms co-condensate with SGS3, another intrinsically disordered protein (IDP). The co-condensation is critical for PTGS as U1-70k LC variants display reduced SGS3 co-condensation, loss of RNA loading onto SGS3, and impaired RNAi function. Taken together, U1-70k channels nascent RNA to SGS3/FVE via its LLPS action to enter PTGS pathway.

molecular biology↗

Construction of Complete Telomere-to-Telomere Genome Assembly of the Rabbit Using Haploid Embryonic Stem Cells

The rabbit owns commercial importance in meat and fur production, and also has long served as a valuable animal model in biomedical research. Yet, the complete assembly of a high-quality rabbit reference genome has not been established. Here, we present a telomere-to-telomere (T2T) genome assembly of the New Zealand White (NZW) rabbit, the most complete and accurate rabbit genome to date. Using a haploid embryonic stem cells (haESCs) to overcame the challenges of homologous sequence interference and structural variation, we generate a genome with a contig N50 of 137.71 Mb, anchoring all chromosomes with only three gaps, through integrating long-read sequencing (PacBio HiFi, Oxford Nanopore), Hi-C, and Illumina data. Quality assessments reveal near-complete coverage of the BUSCO mammalian gene set, with 99.61% completeness. Notably, this complete assembly allowed for the first comprehensive annotation of the rabbit immunoglobulin loci and major histocompatibility complex region, which were previously unresolved in earlier genome versions. The NZW-T2T assembly based on haESCs not only provides a critical resource for advancing rabbit-based research, but also set a new benchmark for de novo genome assembly for other animals with complex genome.

genomics↗

Exploring Erdr1-Mid1 Axis: Shared Risk Factors and Molecular Mechanisms in Aging and Degenerative Diseases

Aging and degenerative diseases are characterized by the progressive decline in cellular, tissue, and organ function, resulting in a significant reduction in quality of life and posing major medical challenges. This highlights the urgent need to elucidate the underlying mechanisms and to develop innovative therapeutic approaches. In this study, we identify Erdr1 and Mid1 as shared risk factors for aging and multiple degenerative diseases. We propose that they contribute to disease progression by modulating oxidative stress, a well-established driver of aging and degenerative processes. We demonstrate that Erdr1 and Mid1 are both involved in oxidative stress regulation. Notably, Erdr1 undergoes alternative splicing in response to oxidative stress, resulting in reduced production of its antioxidant isoforms (Erdr1-177 and Erdr1-209), while promoting the secretion of its pro-oxidant isoform (Erdr1-145). Moreover, Erdr1-145 exacerbates oxidative damage by activating Mid1, a key inducer of oxidative stress. The Erdr1-Mid1-oxidative stress axis provides a molecular mechanistic basis for their shared role as risk factors for aging and degenerative diseases. Furthermore, we propose therapeutic strategies to mitigate cellular damage by regulating Erdr1 levels, implying a straightforward and effective approach for in situ repair of damage associated with aging and degenerative diseases. HighlightsO_LIErdr1 and Mid1 are shared risk factors in aging and multiple degenerative diseases. C_LIO_LIErdr1 and Mid1 contribute to cellular damage and degeneration by modulating oxidative stress. C_LIO_LIErdr1 undergoes alternative splicing in response to oxidative stress, downregulating antioxidant isoforms (Erdr1-177 and 209) while promoting the secretion of prooxidant isoform (Erdr1-145). C_LIO_LIErdr1-145 robustly primes oxidative stress by promoting Mid1 activation, thereby highlighting the role of the Erdr1-Mid1-oxidative stress axis in cellular damage. C_LIO_LIModulating Erdr1 indicates promising strategies for in situ repair of damage associated with aging and degenerative diseases. C_LI

immunology↗

APC loss promotes intestinal transformation through induction of bistable stem cell states

Colorectal cancer (CRC) is a leading cause of cancer deaths, predominantly initiated by genetic inactivation of the APC tumor suppressor. Current dogma holds that APC inactivation promotes tumorigenesis via hyperactivation of the WNT pathway transcriptional effector, {beta}-CATENIN. Although {beta}-CATENIN activation is required for tumor initiation, activating mutations in {beta}-CATENIN are infrequent. Here, we ask what underlies the selective pressure for APC inactivation by comparing the oncogenic effects of APC loss to {beta}-CATENIN hyperactivation. We find that APC loss activates a {beta}-CATENIN-independent fetal intestinal transcriptional program driven by dysregulation of GSK-3 activity upon the LIM-domain protein AJUBA, a positive regulator of YAP. This results in AJUBA stabilization and downstream transcriptional activation of the YAP-driven fetal intestinal gene expression program. We find that {beta}-CATENIN- and YAP-driven transcriptional states are mutually exclusive, existing in an interchangeable bistable balance among APC-null cells. This results in more robust tumor initiation and metastatic progression downstream of APC loss relative to {beta}-CATENIN activation. Taken together, our findings explain the preferential selection for APC inactivation in CRC development and illuminate how {beta}-CATENIN- and YAP-driven gene expression programs coexist to promote tumorigenesis.

cancer biology↗

Functional, Immunogenetic, and Structural Convergence in Influenza Immunity between Humans and Macaques

Human B cell immunity to the influenza hemagglutinin (HA) stem region, a universal influenza vaccine target, is often stereotyped and immunogenetically restricted, posing challenges for study outside humans. Here, we show that macaques vaccinated with a HA stem immunogen elicit human-like public B cell lineages targeting two major conserved sites of vulnerability, the central stem and anchor epitopes. Central stem antibodies were predominantly derived from VH1-138, the macaque homolog of human VH1-69, a VH-gene preferentially used in human central stem broadly neutralizing antibodies (bnAbs). Similarly, macaques produced anchor bnAbs with the human-like NWP motif. Both bnAb lineages were functionally and structurally analogous to their human counterparts, with recognition mediated largely by germline-encoded motifs. Thus the macaque immunoglobulin repertoire supports human-like public bnAb responses to influenza HA. Moreover, this underscores the utility of homologous germline-encoded immunity, suggesting that immune repertoires of macaques and humans may have been similarly shaped during evolution. HIGHLIGHTS[bullet] Functional human-like public antibody lineages can be elicited to HA stem supersites in macaques. [bullet]Macaque central stem bnAbs are predominantly derived from VH1-138, a VH-gene homologous to human VH1-69. [bullet]The human-like CDR L3 NWP anchor epitope-targeting lineage can be elicited in macaques. [bullet]Central stem and anchor bnAbs from humans and macaques engage their respective epitopes with atomic level similarity.

immunology↗

Exploring Hippocampal Vulnerability: Diminished Angiogenic Capacity in the Hippocampus Compared to the Cortex

BackgroundThe hippocampus is one of the first regions affected in various neurodegenerative diseases. In this study, investigated the vascular factors contributing to its susceptibility, aiming to elucidate the underlying vascular mechanisms. MethodUtilizing publicly available single-cell databases, we analyzed the differential expression of genes in blood-brain barrier (BBB)-associated cells within the hippocampus and compared them to those in the cortex. Those genes were further validated in mouse and ischemia rat models. ResultsWe identified differentially expressed genes (DEGs) in endothelial cells, pericytes, and astrocytes in the BBB. Subsequent gene ontology (GO) enrichment analysis and protein-protein interaction (PPI) network analysis identified key hub genes: Kdr, Fn1, Pecam1, Cd34, and Cd93, that related to angiogenesis. They differential expression was then experimentally verified using micro-vessels from mouse and rat brains: In the rat ischemia model, we observed up-regulation of angiogenesis-related genes, including Kdr, Cd34, and Cd93, in the microvasculature of both the hippocampus and cortex, with relatively lower expression in the hippocampus. ConclusionThese findings suggest that the hippocampus has a reduced angiogenic capacity compared to the cortex, which may contribute to its increased vulnerability to neurological disorders.

neuroscience↗

Coro1a promotes the multivesicular body and plasma membrane fusion by facilitating PKM2-mediated SNAP-23 phosphorylation

During the formation of endosomal pathway-dependent extracellular vesicles (EVs), intraluminal vesicles in multivesicular bodies (MVBs) fuse with the plasma membrane (PM), releasing intraluminal vesicles to produce exosomes. Soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) complex, membrane fusion effector, is essential in this process. However, which SNARE complex is involved in MVB and PM fusion and how its assembly is regulated remain elusive. We have demonstrated that neddylation of Coro1a inhibits EV secretion by inducing Rab7-mediated degradation of MVBs. Here, we find that Coro1a increases EV biogenesis by promoting the assembly of SNARE complex STX-12-SNAP-23-VAMP-7 in a neddylation-independent manner. Coro1a activates PKM2 to enhance SNAP-23 phosphorylation in the presence of neddylation inhibitor MLN4924, which drives SNAP-23 to recruit more STX-12 and VAMP7, leading to increased formation of the STX-12-SNAP-23-VAMP-7 complex. Correspondingly, Coro1a-induced EV biogenesis is eliminated by PKM2 inhibitor or SNAP-23 silencing independent of neddylation. Reduced EVs from tumors due to Coro1a knockout cannot be observed in MLN4924-treated tumor mice with PKM2 inhibitor treatment either. Furthermore, Coro1a increases in tumor tissues from lung tumor patients with tumor progression, and high Coro1a indicates unfavorable survival of tumor patients, suggesting that increased Coro1a-mediated enhanced EV production from tumors accelerates tumor progression. Altogether, our data demonstrate that Coro1a facilitates EV biogenesis by promoting the assembly of STX-12-SNAP-23-VAMP-7 complex during MVB and PM fusion independent of neddylation, thereby being involved in the progression of EV-related diseases.

cell biology↗