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: 10765844
• 负责人: Parker C. Wilson
• 金额: $ 16.79万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-01-01 至 2026-12-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10765844
• 关键词: ATAC-seq; 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 Services; Communication; Crossbreeding; 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; LoxP-flanked allele; 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; Transcript; Universities; Up-Regulation; Urine; Washington; Work; Writing; XCL1 gene; candidate identification; career development; cell type; chromatin modification; clinical training; 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; single nucleus RNA-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.

项目概要/摘要 糖尿病肾病是终末期肾病的主要原因，也是发病的主要原因 我们已成功进行单核 RNA 测序 (snRNAseq) 和单核 RNA 测序 (snRNAseq)。 对五个健康对照和八个糖尿病肾脏样本进行核 ATAC 测序 (snATACseq) 进行测量 早期人类糖尿病肾的细胞类型特异性转录和染色质构象图谱 糖尿病近曲小管中差异表达的转录本显示上调。 该提案的目的是糖异生基因和皮质类固醇信号通路的富集。 整合 snRNAseq 和 snATACseq 以确定染色质可及性是否发生变化 调节糖异生的皮质类固醇敏感基因的增强子和启动子区域。 随后用糖尿病损伤的体外模型和糖尿病小鼠模型验证了我们的体内研究结果。 该提案建立在主要研究者之前的研究经验和临床培训的基础上。 目前，Parker Wilson 博士将 25% 的时间花在肾脏和分子病理学临床服务上 其余 75% 分配给 Benjamin Humphreys 博士的实验室进行基础研究。 与 Humphreys 博士建立了指导关系，并发表了他对 snRNAseq 数据的分析 人类糖尿病肾脏作为第一作者发表在 PNAS 上 此外，Wilson 博士最近还拥有一位共同第一作者。 Nature 中描述健康成人肾脏中多模式 snRNAseq 和 snATACseq 整合的手稿 这些发现为该应用程序奠定了基础，该应用程序将重点扩展 Dr. 威尔逊在单细胞方法、生物信息学分析和糖尿病肾损伤模型方面的科学技能。 职业发展目标将通过汉弗莱斯博士的指导和顾问来实现 委员会在染色质构象调节、生物信息学和糖尿病肾病方面拥有专业知识。 威尔逊将承担研究伦理、科学传播和资助写作方面的教学课程， 分子生物学和高级计算机编程将进一步拓展他现有的知识库。 会议在华盛顿大学举行，该大学在指导成功的医学科学家方面有着悠久的历史。 完成这一职业发展奖将为威尔逊博士的追求奠定坚实的基础 独立性和 R01 级资金。