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/摘要 糖尿病肾病是终末期肾病的主要原因，也是发病率的主要贡献者 and mortality.我们已经成功地进行了单核RNA测序（snRNAseq）和单个 对五个健康对照和八个糖尿病肾样品进行核ATAC测序（snATACseq）以测量 人早期糖尿病肾脏细胞类型特异性转录和染色质构象谱 疾病糖尿病近曲小管中差异表达的转录本显示， 致瘤基因和参与皮质类固醇信号传导途径的富集。这项建议旨在 整合snRNAseq和snATACseq，以确定在细胞中是否存在染色质可及性的变化。 增强子和启动子区域的皮质类固醇敏感基因，调节胚胎发生。我们将 随后用糖尿病损伤的体外模型和糖尿病小鼠模型验证我们的体内发现。 本提案基于主要研究者之前的研究经验和临床培训。 目前，帕克威尔逊博士将25%的时间用于肾脏和分子病理学临床服务 剩下的75%用于本杰明·汉弗莱斯博士实验室的基础研究。威尔逊博士有一个 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在PNAS。此外，威尔逊博士有一个最近接受的共同第一作者 描述健康成人肾脏中的多模式snRNAseq和snATACseq整合的手稿 通讯这些发现为这项应用提供了基础，这项应用将集中在扩展Dr. Wilson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。 该委员会在染色质构象调节、生物信息学和糖尿病肾病方面具有专长。博士 威尔逊将从事研究伦理、科学传播和资助写作方面的教学课程， 分子生物学和先进的计算机编程，以进一步他现有的知识基础。这项工作将 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建立一个坚实的基础，威尔逊博士，因为他追求 独立性和R 01级资金。