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.

项目摘要/摘要 糖尿病性肾脏疾病是终末期肾脏疾病的主要原因，也是发病率的主要原因 和死亡率。我们已经成功地进行了单核RNA测序（SNRNASEQ）和单个 五个健康对照和八个糖尿病肾样品上的核ATAC测序（SNATACSEQ）用于测量 早期人类糖尿病肾脏的细胞型特异性转录和染色质构象谱 疾病。在糖尿病近端细胞中表现出不同表达的转录本的上调 糖原性基因和涉及皮质类固醇信号传导的途径富集。该建议旨在 集成的SNRNASEQ和SNATACSEQ，以确定该染色质可访问性是否有变化 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。我们将 随后，使用糖尿病损伤的体外模型和糖尿病小鼠模型来验证我们的体内发现。 该建议是基于主要研究者先前的研究经验和临床培训的基础。 目前，帕克·威尔逊（Parker Wilson）博士将他的时间的25％用于肾脏和分子病理学临床服务 在本杰明·汉弗莱斯（Benjamin Humphreys）博士的实验室中，其余75​​％分配给基础研究。威尔逊博士有一个 与Humphreys博士建立了心理关系，并发表了他对SNRNASEQ数据的分析 人类糖尿病肾脏是PNAS的第一作者。此外，威尔逊博士最近获得了联合第一作者 在自然界健康的成人肾脏中描述多模式SNRNASEQ和SNATACSEQ集成的手稿 通讯。这些发现为本申请提供了基础，该应用程序将着重于扩展博士。 威尔逊在单细胞方法中的科学技能，生物信息学分析和糖尿病肾损伤模型。 Humphreys博士和咨询公司将通过心理实现职业发展目标 委员会具有调节染色质构象，生物信息学和糖尿病性肾病的专业知识。博士 威尔逊将在研究伦理学，科学沟通和授予写作中进行教学课程， 分子生物学和高级计算机编程，以进一步发展他的现有知识基础。工作将 在华盛顿大学（华盛顿大学）有着悠久的成功身体科学家的历史。 该职业发展奖的完成将为威尔逊博士追求的坚实基础 独立和R01级资金。