Biology subjects

Yang, H.

Publications and source records attributed to Yang, H..

At least 217 records · Page 12Linked to original sources

bioRxiv2024

Geographic-specific Diversification of Vaginal Microbiota in China

The vaginal microbiota is pivotal to womens reproductive health, with significant geographic and ethnic heterogeneity suggesting evolutionary adaptations to environmental, climatic, and lifestyle factors. In this multi-centric study, we analyzed vaginal microbiome samples from women across five provinces in China, each characterized by distinct climatic, ethnic, and lifestyle traits. Our results indicate that the vaginal microbiota among Chinese women is characterized by low alpha diversity, with Lactobacillus crispatus being the dominant species in Community State Type (CST) I, especially in the northern regions. While most participants exhibited stable CSTs throughout their menstrual cycles, fluctuations were observed in some individuals. We also detected low-abundance bacterial species such as Fannyhesea vaginae and Sneathia vaginalis. Comparative genomic analysis of high-quality metagenome-assembled genomes of L. crispatus revealed minimal single nucleotide polymorphism (SNP) variations within individuals but substantial diversity between individuals, delineating a distinct separation between vaginal-associated populations (VAP) and gut-associated populations (GAP). These findings highlight the intricate dynamics of the vaginal microbiome, which are markedly influenced by geographic and individual determinants. This study offers an in-depth phylogenetic and genomic analysis of the vaginal microbiota among Chinese women, underscoring the imperative for further research into the impact of regional and individual variations on microbial community composition. Such knowledge could facilitate the development of personalized vaginal health management strategies that accommodate the unique microbiome profiles specific to different geographical regions. ImportanceThis study provides a groundbreaking analysis of the vaginal microbiota among Chinese women, offering insights into the population structure and evolutionary dynamics of this critical component of womens reproductive health. The research reveals that Lactobacillus crispatus, a key species in vaginal health, has a recent evolutionary emergence and exhibits significant geographic variation in prevalence, being more common in northern China. This discovery, along with the identification of distinct vaginal-associated and gut-associated populations, underscores the importance of understanding the vaginal microbiotas complexity. The findings have profound implications for developing personalized strategies to manage vaginal health, tailored to regional microbiome characteristics. This work not only advances our understanding of the vaginal microbiotas role in health and disease but also sets the stage for future research into the factors that shape microbial community structures across diverse populations.

microbiology↗

bioRxiv2024

RING1 dictates GSDMD-mediated inflammatory response and host susceptibility to pathogen infection

RING1 is an E3 ligase component of the polycomb repressive complex 1 (PRC1) with known roles in chromatin regulation and cellular processes such as apoptosis and autophagy. However, its involvement in inflammation and pyroptosis needs to be better characterized. Here, we found that human RING1, but not RING2, promoted K48-linked ubiquitination of Gasdermin D (GSDMD) and acted as a negative regulator of pyroptosis and bacterial infection. Ring1 knockout mice increased the bacterial loads and mortality rate by treatment of S. typhimurium. Upon infection by M. tuberculosis (Mtb) H37Rv strain, RING1 deletion initially reduced bacterial loads but later increased lung inflammation and impaired immune defense responses in mice. Also, Ring1 knockout exacerbated acute sepsis induced by lipopolysaccharide (LPS). Mechanistically, RING1 directly interacts with GSDMD and ubiquitinates the K51 and K168 sites of GSDMD for K48-linked proteasomal degradation, thereby inhibiting pyroptosis. Inhibition of RING1 E3 ligase activity by mutation or small molecular compound increased GSDMD level and cell death during pyroptosis. Our findings reveal that RING1 dictates GSDMD-mediated inflammatory response and host susceptibility to pathogen infection, highlighting RING1 as a potential therapeutic target for combating infectious diseases. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=115 SRC="FIGDIR/small/622030v1_ufig1.gif" ALT="Figure 1"> View larger version (32K): org.highwire.dtl.DTLVardef@e6d4a6org.highwire.dtl.DTLVardef@92f137org.highwire.dtl.DTLVardef@11a69c4org.highwire.dtl.DTLVardef@115dc0b_HPS_FORMAT_FIGEXP M_FIG Graphic Abstract C_FIG HighlightsO_LIE3 ubiquitin ligase RING1 targets GSDMD for proteasomal degradation. C_LIO_LIMice lacking RING1 are more vulnerable to S. typhimurium infection and LPS- induced sepsis. C_LIO_LIIn the early stages of M. tuberculosis (Mtb) infection, Ring1-/- mice exhibit efficient bacterial clearance. C_LIO_LIChronic inflammation in Ring1-/- mice impairs the ability to control Mtb in the late stages of infection. C_LI

cell biology↗

bioRxiv2024

Reinforced identity confers cells with cancer hallmarks

Genetic alterations trigger tumorigenesis in a cell type-specific manner, but the biological basis for this, remains unclear. Here we define reinforced original cell identity established by interaction between genetic alterations and original cell type is a pivotal determinant of tumor development. Using AML-M5MLL-AF9 as a model system, we demonstrated that MLL-AF9 enhanced key transcriptional factors (TFs) expression of promonocyte and cooperated with them to amplify their downstream programs for AML hallmarks. Interactions between MLL-AF9 and promonocyte endowed cancer cells with a reinforced promonocyte identity. Reinforced intrinsic cell identity also sustained cancer hallmarks across cancer types. Furthermore, we identified narciclasine, as an effective disruptor of reinforced promonocyte identity, effectively treating AML-M5MLL-AF9. This study not only identified reinforced cell identity as a fundamental tumor driver beyond genetic mutations but also offers a proof-of-concept for reinforced identity- targeted cancer therapy.

cancer biology↗

bioRxiv2024

Evaluation of a Functional Portable Hemodialysis Machine, XKIDNEY

Chronic kidney failure and end-stage renal disease (ESRD) are increasing globally, but home hemodialysis offers a critical means of improving patient quality of life and reducing national healthcare costs. This study investigates the performance of a portable hemodialysis (HD) device named XKIDNEY. The efficacy and safety of the device were determined with focus on solute clearance, fluid balancing accuracy, and blood cell integrity in an ex-vivo test environment. A comparison of the performance of XKIDNEY and a conventional HD unit showed that XKIDNEY had superior balancing accuracy without any technical malfunction. In addition, low molecular weight solutes, such as urea and creatinine, were effectively removed. These findings demonstrate that XKIDNEY enhances treatment portability without compromising performance and indicate XKIDNEY is a feasible home hemodialysis option.

bioengineering↗

bioRxiv2024

Meditation in the third-person perspective modulates minimal self and heartbeat-evoked potentials

Experienced meditation practitioners often report altered states of their sense of self, including decentering and distancing the self from the body and ones current concerns. Altered states of the sense of self, such as disembodiment and distancing of the self from the body, have also been induced experimentally using virtual reality (VR) and linked neurally to heartbeat evoked potentials (HEPs). Whereas many studies investigated the related neural correlates of such decentering during meditation, none experimentally modulated the sense of self during meditation practice using VR nor determined the potentially associated behavioral changes of the sense of self. Here we determined HEPs and behavioral measures in 23 participants who performed a guided meditation practice in VR, either from a third-person (3PP) or first-person perspective (1PP) to modulate the sense of self. In the 3PP versus 1PP meditation condition, we report stronger sensations of detachment and disconnection, reduced salience of the perceived body boundary, and reduced self-identification with the body. HEP analysis revealed differential neural responses between conditions, characterized by a more negative HEP amplitude in the 3PP condition, associated with activation of the posterior cingulate cortex and medial prefrontal cortex. Leveraging a new VR-supported meditation platform and methods, these data link the sense of self in meditation practice to the neuroscience of the bodily self, based on subjective, behavioral, and neural measures.

neuroscience↗

bioRxiv2024

Dysregulated balance of D- and L-amino acids modulating glutamatergic neurotransmission in severe spinal muscular atrophy.

Spinal muscular atrophy (SMA) is a neuromuscular disorder caused by reduced expression of the survival motor neuron (SMN) protein. In addition to motor neuron survival, SMN deficiency affects the integrity and function of afferent synapses that provide glutamatergic excitatory drive essential for motor neuron firing and muscle contraction. However, it is unknown whether deficits in the metabolism of excitatory amino acids and their precursors contribute to neuronal dysfunction in SMA. To address this issue, we measured the levels of the main neuroactive D- and L-amino acids acting on glutamatergic receptors in the central nervous system of SMN{Delta}7 mice as well as the cerebrospinal fluid (CSF) of SMA patients of varying severity before and after treatment with the SMN-inducing drug Nusinersen. Our findings reveal that SMN deficiency disrupts glutamate and serine metabolism in the CSF of severe SMA patients, including decreased concentration of L-glutamate, which is partially corrected by Nusinersen therapy. Moreover, we identify dysregulated L-glutamine to L-glutamate conversion as a shared neurochemical signature of altered glutamatergic synapse metabolism that implicates astrocyte dysfunction in both severe SMA patients and mouse models. Lastly, consistent with a correlation of higher CSF levels of D-serine with better motor function in severe SMA patients, we show that daily supplementation with the NMDA receptor co-agonist D-serine improves neurological deficits in SMN{Delta}7 mice. Altogether, these findings provide direct evidence for dysregulation of D- and L-amino acid metabolism linked to glutamatergic neurotransmission in severe SMA and have potential implications for treating this neurological disorder.

neuroscience↗

bioRxiv2024

A novel PhoPQ-potentiated mechanism of colistin resistance impairs membrane integrity in Pseudomonas aeruginosa

Increasing bacterial resistance to colistin, a vital last-resort antibiotic, is an urgent challenge. We previously reported that magnesium sequestration by Candida albicans enables Pseudomonas aeruginosa to become colistin-resistant. Here, we show that Mg{superscript 2} depletion drives P. aeruginosa to evolve greater colistin resistance through genetic changes in lipid A biosynthesis-modification pathways and a putative magnesium transporter. These mutations synergize with the Mg2+-sensing PhoPQ two-component signaling system to remodel lipid A structures of the bacterial outer membrane in previously uncharacterized ways. One predominant mutational pathway relies on early mutations in htrB2, a non-essential gene involved in lipid A biosynthesis, which enhances resistance but compromises outer membrane integrity, resulting in fitness costs and increased susceptibility to other antibiotics. A second pathway achieves increased colistin resistance independently of htrB2 mutations without compromising membrane integrity. In both cases, reduced binding of colistin to the bacterial membrane underlies resistance. Our findings reveal that Mg2+ scarcity unleashes two novel trajectories of colistin resistance evolution in P. aeruginosa. (160)

microbiology↗

bioRxiv2024

Pan-Prediction of MHC-II Restricted Epitopes Across Species via an Alphafold-based Quantification Scheme

Predicting MHC-II restricted epitopes across species used to be challenging, but Alphafold (AF) may provide a structure-based pan-prediction solution. In this study, we established the new tool AF-pred with a clear standard for quantitative prediction results. Compared to the sequence-based tools heavily trained with human ligandome, AF-pred does not show advantage in predicting the binding patterns of human HLA-II but has far better performance in predicting binding patterns of other animals MHC-II. Using recently resolved bat MHC-II structures, we analyzed AF-preds prediction capability, logic and limitation. In addition, we also explored the impact of AF algorithm iterations on the prediction of MHC-II restricted epitopes. The results demonstrated that AF-pred is capable of cross-species prediction of MHC-II restricted epitopes and is conducive to the development of novel veterinary vaccines.

immunology↗

bioRxiv2024

Whole-Process 3D ECM-Encapsulated Organoid-Based Automated High-Throughput Screening Platform Accelerates Drug Discovery for Rare Diseases

The use of organoids, especially patient-derived organoids, for high-throughput screening (HTS) is widely accepted due to their ability to mimic the three-dimensional (3D) structure, function, and drug responses of in vivo tissues. However, the complexity of handling extracellular matrix (ECM) components with traditional HTS devices leads to the utilization of suspension cultures in matrix-free or matrix-low conditions during HTS, which can alter their transcriptomic landscape and drug responses. Here, we develop a whole-process 3D ECM-encapsulated organoid-based automated HTS (wp3D-OAHTS) platform, which enables the rapid and accurate generation of uniformly distributed 3D cell-matrix mixture domes at the center of each well in 96-well plates. This approach replicates the process of manual organoid culture but with superior stability and reproducibility. Utilizing this platform, we screened 2,802 compounds on neuroendocrine cervical cancer organoids, a rare malignancy with significant unmet clinical needs. We identified 7 top hits that display strong anti-tumor effects with remarkably low half-maximal inhibitory concentration (IC50) and validated the in vivo efficacy of Quisinostat 2HCl. Additionally, we demonstrated that employing 3D ECM-encapsulated organoid cultures for HTS, rather than suspended cultures, provides optimal conditions for drug discovery. Our wp3D-OAHTS platform significantly improves the rapidity and efficiency of new drug discovery for rare diseases.

bioengineering↗

bioRxiv2024

Genomic stability in the Galapagos Scalesia adaptive radiation: Consistent transposable element accumulation despite hybridization and ecological niche shifts

Transposable elements (TEs) have been hypothesized to play a pivotal role in driving diversification by facilitating the emergence of novel phenotypes and the accumulation of divergence between species. Hybridization and adaptation to novel niches are hypothesized to influence TE accumulation in genomes by disrupting genomic regulation, including the suppression of TE replication. The rapid speciation and ecological diversification characteristic of adaptive radiations offer a unique opportunity to examine the link between TE accumulation and speciation, diversification, hybridization and adaptation. Here, focusing on all 15 species of the genus Scalesia (Asteraceae), a radiation endemic to the Galapagos Islands, we test whether hybridization or shifts in ecological niche are associated with changes in TE accumulation in genomes. Our analyses reveal little to no variation in TE accumulation among Scalesia species nor its hybrid populations. Shifts in ecological niches, linked to climatic variation, did not result in discernible changes in TE accumulation, a surprising finding given the anticipated selective pressure imposed by aridity, a factor often linked to genome size reduction. We found no distinct patterns in the temporal accumulation of TEs, and no effects at the class or superfamily level. Our findings challenge the assertion that TEs have directly driven diversification, speciation and local adaptation. Instead, we suggest that TEs may simply be along for the ride, rather than actively contributing to plant diversification.

evolutionary biology↗

bioRxiv2024

Characterization of Autophagy Cargo Profiles during Starvation and Exercise with a Novel Autophagosome Isolation Method

Autophagy is a cellular process to clear unwanted and dysfunctional cellular cargoes, which are sequestered in autophagosomes before their delivery to lysosomes for degradation. Autophagy cargo selection, mediated by cargo receptors, varies across cell types and conditions. Understanding the cargo features is essential for elucidating autophagys function in specific physiological or pathological contexts. Here we present a simple and rapid method for isolating LC3B-positive autophagosomes from the tissues of GFP-LC3 transgenic mice, a widely used autophagy reporter model, without relying on the complex ultracentrifugation steps required by traditional methods. When combined with quantitative proteomics, this approach enables efficient in vivo characterization of autophagy cargoes. We applied this method to establish autophagy cargo profiles in skeletal muscle during starvation and exercise, two physiological conditions that activate autophagy, and identified distinct cargo selection patterns, with significantly higher levels of ER-phagy and ribophagy observed during starvation. We further revealed the ER-phagy receptors TEX264 and RETREG1/FAM134B as potential mediators of the elevated ER-phagy under starvation. In summary, we report an efficient workflow for in vivo autophagy cargo characterization and provide detailed analysis and comparison of cargo profiles under starvation and exercise conditions.

cell biology↗

bioRxiv2024

Transcriptional memories mediate the plasticity of sulfide stress responses to enable acclimation in Urechis unicinctus

To cope with environmental stresses, organisms often adopt a memory response upon primary stress exposure to facilitate a quicker and/or stronger reaction to recurring stresses. Somatic stress memory is essential in dealing with contemporary stress. The earliest sign of somatic stress memory is a change in gene transcription levels, which alters physiology and phenotype to better cope with stress. Sulfide is a common environmental pollutant; however, some organisms have successfully colonized sulfur-rich environments. Whether stress memory plays important role in sulfide stress adaptation remains unclear. In this study, to determine whether Urechis unicinctus, a sulfur-tolerant organism, retains the memory of previous sulfide stress, we simulated a repetitive sulfide stress/recovery system. The results showed that the tolerance of U. unicinctus to sulfide stress was significantly increased after priming with 50 {micro}M sulfide. Further, transcriptional memory genes (TMGs) involved in regulating sulfide stress memory were identified, classified according to their expression patterns, and functionally analyzed. TMGs involved in sulfide metabolism, sugar metabolism, and protein homeostasis pathway showed an enhanced response, whereas those related to DNA repair pathway demonstrated a modified response pattern. Our study indicated that U. unicinctus retains memory of sulfide stress priming, which mediates plasticity to accelerate sulfide stress adaptation.

zoology↗

bioRxiv2024

FishCODE: a web-based information platform for comprehensive omics data exploration in fish research

In terms of the utilization of omics data, the current fish database analysis functions are primarily relatively simple tools at the transcriptional level, aimed at obtaining the co-expression levels of specified genes or the data visualization of multiple genes, and do not enable users to perform comprehensive omics data analysis. Furthermore, the gene-level information currently provided by these multispecies fish genomics databases is incomplete, and there is a lack of a comprehensive portal that can offer multidimensional genetic information. To address these challenges, we collected extensive multi-omics information on 35 fishes and established the primary comprehensive multi-omics data information platform for fish, FishCODE (http://bioinfo.ihb.ac.cn/fishcode). We have collected experimental background of dataset which pertaining to the target fishes, selected a range of datasets that encompass a broad spectrum of research areas, and downloaded the corresponding raw omics data from public repositories such as the Sequence Read Archive (SRA). Through a unified pipeline analysis, FishCODE contains 11,216 samples from 540 sets of genomic, transcriptomic, and methylomic datasets. These data encompass transcript structure and expression, gene methylation levels, protein domains, protein subcellular localization, protein interactions, best matched protein (Swiss-Prot), associated SNP site information (47,111,018), orthologous genes, phylogenetic tree and GO/KEGG annotations. To facilitate comparison, we annotated the experimental background data sets of the FishCODE, FishGET, PhyloFish, FishSED and FishSCT databases using the Fish Experimental Condition Ontology. Currently, the FishCODE database omics dataset includes 146 unique experimental condition words, 654 cumulative experimental condition words, and 13 species with rich experimental background (more than 20 unique FECO words). These data are 3.5 times (42), 8.3 times (74), and 6.5 times (2) those of the second-ranked databases respectively. We generated word cloud maps for the experimental condition vocabularies of FishCODE and FishGET, illustrating the superior richness of FishCODEs experimental background.

bioinformatics↗

bioRxiv2024

Endocrine persistence in ER+ breast cancer is accompanied by metabolic vulnerability in oxidative phosphorylation

Despite adjuvant treatment with endocrine therapies, estrogen receptor-positive (ER+) breast cancers recur in a significant proportion of patients. Recurrences are attributable to clinically undetectable endocrine-tolerant persister cancer cells that retain tumor-forming potential. Therefore, strategies targeting such persister cells may prevent recurrent disease. Using CRISPR-Cas9 genome-wide knockout screening in ER+ breast cancer cells, we identified a survival mechanism involving metabolic reprogramming with reliance upon mitochondrial respiration in endocrine-tolerant persister cells. Quantitative proteomic profiling showed reduced levels of glycolytic proteins in persisters. Metabolic tracing of glucose revealed an energy-depleted state in persisters where oxidative phosphorylation was required to generate ATP. A phase II clinical trial was conducted to evaluate changes in mitochondrial markers in primary ER+/HER2-breast tumors induced by neoadjuvant endocrine therapy (NCT04568616). In an analysis of tumor specimens from 32 patients, tumors exhibiting residual cell proliferation after aromatase inhibitor-induced estrogen deprivation with letrozole showed increased mitochondrial content. Genetic profiling and barcode lineage tracing showed that endocrine-tolerant persistence occurred stochastically without genetic predisposition. Mice bearing cell line- and patient-derived xenografts were used to measure the anti-tumor effects of mitochondrial complex I inhibition in the context of endocrine therapy. Pharmacological inhibition of complex I suppressed the tumor-forming potential of persisters and synergized with the anti-estrogen fulvestrant to induce regression of patient-derived xenografts. These findings indicate that mitochondrial metabolism is essential in endocrine-tolerant persister ER+ breast cancer cells and warrant the development of treatment strategies to leverage this vulnerability in the context of endocrine-sensitive disease. Statement of SignificanceEndocrine-tolerant persister cancer cells that survive endocrine therapy can cause recurrent disease. Persister cells exhibit increased energetic dependence upon mitochondria for survival and tumor re-growth potential.

cancer biology↗

bioRxiv2024

IGF2BP3 Promotes Autophagy-Mediated TNBC Metastasis via M6A-Dependent, Cap-Independent c-Met Translation

Tumor metastasis involves a series of complex processes and is a major challenge in clinical treatment. The cooperation of epigenetic modifications and metabolic adaptations endows tumor cells with dynamic adaptations for survival in variable environments, which is crucial for tumor metastasis and worth exploring in depth. In this study, we found IGF2BP3 could link epigenetic modification and metabolic adaptation in promoting autophagy-mediated triple-negative breast cancer (TNBC) metastasis. As a TNBC specifically high expressed N6-methyladenosine (m6A) binding protein, IGF2BP3 could bind to the m6A motif of c-Met mRNA, regulating autophagy-mediated epithelial-to-mesenchymal transition (EMT) progress by c-Met/PI3K/AKT/mTOR pathway. Moreover, through the recruitment of eIF4G2 as a collaborator, IGF2BP3 induced the changes in c-Met protein expression not by affecting mRNA stability, but by facilitating its mRNA translation initiation in an m6A-dependent and cap-independent manner. In conclusion, our study extended the role of IGF2BP3 in autophagy mediated TNBC metastasis, found IGF2BP3 could bind to the m6A motif on the 5', 3'-UTR of c-Met mRNA to facilitate its translation in a cap-independent manner.

cancer biology↗

bioRxiv2024

Tissue-specific and functional loci analysis of CASP14 gene in the sheep horn

Under the current context of intensive farming, small-horned animals are more suitable for large-scale breeding. The CASP14 gene, closely associated with skin and keratin formation, may influence horn size due to its link with skin development. This study comprehensively analyzed the tissue-specific expression of CASP14 using RNA-Seq data, identified functional sites through whole-genome sequencing (WGS), and investigated allele-specific expression (ASE) validated by KASP assays. Results showed significantly higher CASP14 expression in the scurred group com-pared to the SHE group, with pronounced expression in the skin. Interbreed comparisons also revealed elevated CASP14 levels in the scurred group. Analysis of potential functional sites indicated structural similarities in the CASP14 protein among horned mammals. Five ASE events were discovered, and intersecting these with SNPs and high fixation index loci (Fst > 0.05) identified three potential functional sites: 7941628, 7941817, and 7941830. The SNP site 7944295 was selected for T-test analysis and further validated through KASP assays, corroborating the role of CASP14 in sheep horn phenotypes. Our findings suggest that CASP14 plays a significant role in horn development, offering insights into breeding strategies for small-horned animals.

zoology↗

bioRxiv2024

Phytop: A tool for visualizing and recognizing signals of incomplete lineage sorting and hybridization using species trees output from ASTRAL

Incomplete lineage sorting (ILS) and introgression/hybridization (IH) is prevalent in nature and thus frequently result in discrepancies within phylogenetic tree topologies, leading to misinterpretation of phylogenomic data. Despite the availability of numerous tools for detecting ILS and IH among species, many of these tools are lacking effective visualization, or are time-consuming, or require prior predetermination. Here, we addressed these shortcomings by developing a fast-running, user-friendly tool called Phytop. By defining ILS and IH indices to quantify ILS and IH, this tool can detect the extent of ILS and IH among lineages with high reliability, and can visualize them based on the gene tree topology patterns constructed using ASTRAL. We tested Phytop extensively using both simulated and real data, and found that it enables users to quickly and conveniently estimate the extent of ILS and IH, thus clarifying the phylogenetic uncertainty. Phytop is available at https://github.com/zhangrengang/phytop and is expected to conveniently contribute to the intuitive inference of genetic relationships among lineages in future research.

bioinformatics↗

bioRxiv2024

SIRT5-mediated GLS and GDH desuccinylation attenuates autophagy in MAC-T cells induced by ammonia

Our previous research revealed that NH3 regulated autophagy dependent on SIRT5 in MAC-T cells. Interestingly, SIRT5 reduced the content of NH3 and glutamate by inhibiting GLS activity, ADP/ATP value also declined. In this study, SIRT5 interacted with endogenous GLS and GDH, and had no effect on endogenous GLS and GDH expression. SIRT5 declined significantly the succinylation levels of GLS and GDH, and further reduced the enzymatic activity of GLS and GDH. SIRT5 declined the glutamine metabolism, which attenuated ammonia release in MAC-T cells, accompanying with cellular autophagy decline, reducing the formation of autophagosome. Deletion of SIRT5 increased the content of NH3 and glutamate, as well as promotes autophagy, which could be alleviated by SIRT5 overexpression. SIRT5 KO was associated with increased succinylation and activity of GLS and GDH, as well as autophagy response in MAC-T cells. Furthermore, SIRT5 promoted the maintenance of mitochondria homeostasis. Mechanistically, SIRT5 modulated the succinylation levels and enzymatic activities of GLS and GDH in mitochondria and promoted the maintenance of mitochondria homeostasis, further attenuating ammonia-stimulated autophagy in MAC-T cells. HighlightsO_LISIRT5 catalyzed lysine desuccinylation of GLS and GDH. C_LIO_LIGLS and GDH enzymatic activity were enhanced by lysine succinylation. C_LIO_LIGLS and GDH were required for SIRT5 to regulate ammonia-induced cellular autophagy. C_LIO_LISIRT5 promoted the maintenance of mitochondrial homeostasis C_LI

physiology↗