The Single Cell Landscape of Early Human Diabetic Nephropathy

早期人类糖尿病肾病的单细胞景观

• 批准号: 10368354
• 负责人: Parker C. Wilson
• 金额: $ 16.78万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-01-01 至 2022-12-02
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10368354
• 关键词: Adrenal Cortex Hormones; Adult; Advisory Committees; Albuminuria; Antihypertensive Agents; Atlases; Basic Science; Binding Sites; Bioinformatics; Biological Markers; Blood Glucose; Body Weight; Cell Nucleus; Cells; ChIP-seq; Chromatin; Chronic; Chronic Kidney Failure; Clinical; Clinical Services; Communication; DNA; Data; Data Set; Dexamethasone; Diabetes Mellitus; Diabetic Nephropathy; Diabetic mouse; Disease; Disease Progression; Dissociation; End stage renal failure; Enhancers; Enterobacteria phage P1 Cre recombinase; Epithelial Cells; Exons; Foundations; Funding; Gene Expression; Genes; Genetic Transcription; Glucocorticoid Receptor; Glucocorticoids; Gluconeogenesis; Glucose; Goals; Grant; Human; Hyperglycemia; Immunofluorescence Immunologic; Immunohistochemistry; Impairment; In Vitro; Individual; Inflammation; Injury; Injury to Kidney; Insulin; K-Series Research Career Programs; Kidney; Knock-out; Laboratories; Manuscripts; Measures; Memory; Mentors; Metabolic; Metabolism; Methods; Modeling; Molecular Biology; Molecular Conformation; Morbidity - disease rate; Mus; Nature; Nuclear; Output; Pathway interactions; Patients; Physicians; Principal Investigator; Process; Promoter Regions; Publishing; Recording of previous events; Regulation; Reproducibility; Research; Research Ethics; Resources; Response Elements; Sampling; Scientist; Signal Transduction; Site; Solid; TNF gene; Tamoxifen; Testing; Time; Training; Transcript; Universities; Up-Regulation; Urine; Washington; Work; Writing; XCL1 gene; career development; cell type; chromatin modification; computer program; db/db mouse; diabetic; differential expression; experience; genome wide association study; glycemic control; human data; human model; in vitro Model; in vivo; insulin sensitivity; kidney cortex; knowledgebase; molecular pathology; mortality; mouse model; multimodality; multiple omics; non-diabetic; novel; promoter; public database; receptor binding; response; risk variant; single cell sequencing; skills; transcription factor; transcriptome sequencing

Project Summary/Abstract Diabetic kidney disease is the leading cause of end-stage renal disease and a major contributor to morbidity and mortality. We have successfully performed single nucleus RNA sequencing (snRNAseq) and single nucleus ATAC sequencing (snATACseq) on five healthy control and eight diabetic kidney samples to measure the cell-type-specific transcriptional and chromatin conformational profile of early human diabetic kidney disease. The differentially expressed transcripts in the diabetic proximal tubule showed upregulation of gluconeogenic genes and enrichment of pathways involved in corticosteroid signaling. This proposal aims to integrate snRNAseq and snATACseq to determine whether there are changes in chromatin accessibility in the enhancer and promoter regions of corticosteroid-sensitive genes that regulate gluconeogenesis. We will subsequently validate our in vivo findings with an in vitro model of diabetic injury and a diabetic mouse model. This proposal builds on the principal investigator’s previous research experience and clinical training. Currently, Dr. Parker Wilson is spending 25% of his time on the renal and molecular pathology clinical services with the remaining 75% allocated to basic research in Dr. Benjamin Humphreys’ laboratory. Dr. Wilson has an established mentoring relationship with Dr. Humphreys and has published his analysis of snRNAseq data from human diabetic kidney as a first author in PNAS. In addition, Dr. Wilson has a recently-accepted co-first author manuscript describing multimodal snRNAseq and snATACseq integration in the healthy adult kidney in Nature Communications. These findings provide the foundation for this application, which will focus on expanding Dr. Wilson’s scientific skills in single cell methods, bioinformatics analysis and models of diabetic kidney injury. The career development goals will be achieved through mentoring by Dr. Humphreys and an advisory committee with expertise in regulation of chromatin conformation, bioinformatics and diabetic nephropathy. Dr. Wilson will undertake didactic coursework in research ethics, scientific communication and grant writing, molecular biology and advanced computer programming to further his existing knowledgebase. The work will take place at Washington University, which has a strong history of mentoring successful physician-scientists. Completion of this career development award will build a solid foundation for Dr. Wilson as he pursues independence and R01-level funding.

项目总结/文摘