Project description:We employed the Chromium GEM-X single-cell RNA sequencing platform by 10x Genomics to investigate the effects of the GLS1 inhibitor, BPTES, on the kidney single-cell transcriptome of 19- to 20-month-old wild-type male mice. Previous studies have identified GLS1 as a potential senolytic reagent capable of eliminating senescent cells. Our aim was to determine which cell populations are selectively removed by GLS1 inhibition in aged mice and to assess how this intervention alters intercellular interactions across different cell types.
Project description:We employed the Chromium GEM-X single-cell RNA sequencing platform by 10x Genomics to investigate the effects of the GLS1 inhibitor, BPTES, on the lung single-cell transcriptome of 19- to 20-month-old wild-type male mice. Previous studies have identified GLS1 as a potential senolytic reagent capable of eliminating senescent cells. Our aim was to determine which cell populations are selectively removed by GLS1 inhibition in aged mice and to assess how this intervention alters intercellular interactions across different cell types.
Project description:Left ventricular hypertrophy (LVH) is critical in chronic kidney disease (CKD), but its mechanisms remain unclear. This study employed multi-omics approaches, including RNA-seq and integrated metabolomics, to characterize metabolic alterations in cardiomyocytes (CM) of CKD mice, revealing enhanced glutaminolysis and impaired oxidative phosphorylation. We identified that glutaminase 1 (GLS1), the rate-limiting enzyme of glutaminolysis, is significantly upregulated and enzymatically activated in CM under CKD conditions. A multi-center cohort study confirmed that serum-induced GLS1 activity was associated with LV hypertrophic remodeling. Cardiomyocyte-specific deletion or pharmacological inhibition of GLS1 attenuated CKD-induced cardiac hypertrophy. Proteomic analysis revealed that multiple circulating proteins in the serum of CKD patients are associated with GLS1 enzymatic activity. Furthermore, phosphate (Pi), a representative uremic toxin, functions as a critical modulator of GLS1 activity and expression, compromising mitochondrial bioenergetics by promoting ammonia production and reducing mitochondrially encoded respiratory chain proteins. These integrated metabolic effects underscore the pivotal role of GLS1 in orchestrating myocardial energy reprogramming, thereby contributing to CKD-induced LVH.
Project description:To evaluate the roles of glutaminase 1 (GLS1) in vascular smooth muscle cells (VSMCs) phenotypic switching and aortic dissection (AD). Integrative transcriptomic analyses were performed to identify the candidate genes involved in VSMC phenotypic switching in AD. The expression of GLS1 in VSMCs was assessed by qRT-PCR, Western blot and immunofluorescence. RNA-sequencing analysis was performed to recapitulate possible changes in the transcriptome profile of GLS1 in VSMCs. We identified GLS1 as a potential regulator in AD. GLS1 expression was significantly downregulated in VSMCs from both human AD aortic tissues and mouse models. Mechanistically, down-regulation of GLS1 impaired glutamate metabolism, leading to reduced levels of glutathione and α-ketoglutarate, thereby promoting mitochondrial dysfunction and accumulation of reactive oxygen species, which activated the PI3K/AKT pathway and ultimately triggered VSMC phenotypic switching. These findings revealed a critical role of GLS1-mediated glutamate metabolism in VSMC phenotypic switching and suggest a promising therapeutic target for AD.
Project description:Glutaminase (GLS1) is involved in the development of Alcoholic liver disease (ALD). Protein-protein interactions underlie the biological processes impacted by GLS1. we used immunoprecipitation-mass spectrometry (IP-MS) technology and found the proteins interacted with GLS1.
Project description:Here, we performed single cell RNA sequencing (scRNA-seq) of human fetal kidney tissue samples from 2 individual biological specimens (13.7 and 15.4 weeks gestation). The data set is composed of approximately 10,000 cells from diverse renal lineages. Lineages captured include nephron progenitors, epithelium, stroma, immune, and endothelium.
Project description:Human induced pluripotent stem cell-derived kidney organoids have potential for disease modelling and regenerative medicine purposes. However, they lack a functional vasculature and remain immature in in vitro culture. Here, we transplanted kidney organoids at day 7+12 of differentiation in the coelomic cavity of chicken embryos and then compared them to their respective untransplanted controls at d7+13 and d7+20 using scRNAseq and imaging modalities. We demonstrate vascularization and enhanced maturation of transplanted kidney organoids.
Project description:The efficiencies with which activated B lymphocytes proliferate and develop into antibody (Ab)-secreting plasma cells are critical determinants of adaptive humoral immunity and central to sustaining certain autoimmune diseases. Increasing evidence indicates that specific pathways in intermediary metabolism, or their substrate supply, influence lymphocyte differentiation and function. To investigate the role of glutaminolysis by GLS1 and mitochondrial pyruvate import via MPC2 on germinal center B cell differentiaiton and function flow-purified we immunized wild-type, GLS1-KO, MPC2-KO, or double KO mice with SRBC and FACS isolated germinal center B cells after 7 days for transcriptional profiling.