Research Project 2: Molecular analysis of developing post-natal mouse kidney in health and FSGS

研究项目2：健康和FSGS中小鼠产后肾脏发育的分子分析

基本信息

批准号：
10530271
负责人：
MICHAEL I RAUCHMAN
金额：
$ 21.88万
依托单位：
WASHINGTON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-09-21 至 2027-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10530271
关键词：
ATAC-seq Adolescent Adult Affect Age Age-Months Albuminuria Atlases Biological Assay Biopsy Birth Cell Aging Cell Differentiation process Cells ChIP-seq Child Childhood Chromatin Chromatin Remodeling Factor Chromium Clinical Coupled Data Data Set Deacetylase Defect Development Diagnosis Disease Disease Progression Early Diagnosis Embryo End stage renal failure Endowment Epigenetic Process Evolution Failure Fatty acid glycerol esters Focal Segmental Glomerulosclerosis Frequencies Gene Activation Gene Expression Gene Expression Profile Genes Glean Growth Health High Fat Diet Human Hypertension Hypoxia Immune Infant Inflammation Injury Injury to Kidney Kidney Kidney Diseases Kidney Failure Lead Life Lipids Localized Disease Longevity Metabolic Metabolic stress Metabolism Metastasis-associated protein Mission Modeling Molecular Molecular Analysis Morphology Multiomic Data Mus Mutant Strains Mice Neighborhoods Nephrons Nephrotic Syndrome Nucleic Acid Regulatory Sequences Nucleosomes Obesity Pathway interactions Phenotype Physiological Play Public Health Recurrence Regulator Genes Regulatory Element Renal function Renal glomerular disease Repression Research Research Project Grants Resistance Risk Factors Role Signal Pathway Small Nuclear RNA Stress Technology Testing Therapeutic Time Tissues Transcription Factor AP-1 United States National Institutes of Health Urinary tract cell type congenital anomalies of the kidney critical developmental period early adolescence emerging adult epigenomics fetal functional adaptation gene regulatory network genetic risk factor glomerulosclerosis high risk high salt diet human disease human model improved injured insight lipid metabolism loss of function mortality mouse model multiple omics mutant nephrogenesis nephron progenitor novel peer postnatal period potential biomarker repaired senescence stem cells therapeutic target transcriptome sequencing transcriptomics

项目摘要

Summary After birth, the kidney continues to develop for weeks in mice, and for months to years in humans. During this period, the post-natal kidney is undergoing growth and maturation, which requires dynamic changes in gene expression, metabolism, and physiological functions, and is susceptible to insults causing permanent morphological changes and functional adaptation. Post-natal growth and maturation of the kidney play an important role in developmentally programmed diseases. For e.g., children born with a moderate reduction in nephron number may be more sensitive to metabolic stresses, such as high fat or high salt diet, obesity, and renal injury from hypoxia or nephrotoxins. Many studies have generated single cell datasets for fetal and adult mouse kidney. However, in spite of the important role of post-natal insults in development and progression of CKD, post-natal maturation of the kidney has not been comprehensively investigated at the molecular level using state-of-the-art technologies. The lack of a reference atlas for this critical developmental period limits our ability to investigate alterations in cell states and diversity in models of pediatric kidney disease. Glomerular diseases, including nephrotic syndrome, represent an important cause of childhood kidney disease. Focal and segmental glomerulosclerosis (FSGS) is a diagnosis that is increasing in frequency, is often recurrent and treatment resistant, and leads to end stage kidney disease. Secondary glomerulosclerosis and albuminuria develop in common non-glomerular diseases affecting children, such as CAKUT, and portend a higher risk of progression to end stage kidney failure. Currently, there are limited therapeutic options to treat these disorders and a pressing need to better understand disease mechanisms. One of the major mechanisms by which cells respond to stress or injury is by utilizing the chromatin modifying machinery to reprogram gene regulatory networks. We developed a novel mouse model of FSGS due to loss of function of metastases associated protein 2 (Mta2), a core component of the NuRD chromatin remodeling complex. These mutant mice represent an excellent model to investigate how clinically silent defects in nephron endowment and altered cell differentiation can manifest as FSGS in late adolescence/early adulthood. In this proposal we will generate an integrated single cell and spatial transcriptomics mouse atlas (pKidCAP) at post-natal timepoints that coincide with major physiological and developmental changes in the kidney. We will determine the evolution of cell diversity, spatial localization, single cell gene expression signatures and chromatin accessibility states in the developing post-natal kidney in healthy and FSGS tissue. We will apply ChIP-seq and epigenomic editing to test the hypothesis that de-repression of Jun/AP-1 targets in Mta2 mutants promotes inflammation, cell senescence and lipotoxicity. Integration of these datasets with the human pKidCAP atlas (Project 1) will create a unique reference panel to identify pathways uncovered in mouse models that are highly relevant to kidney diseases in children.

概括出生后，小鼠的肾脏会持续发育数周，而人类的肾脏则持续发育数月至数年。期间此时期，产后肾脏正处于生长成熟期，需要动态变化。基因表达、新陈代谢和生理功能，并且容易受到造成永久性损伤的损害形态变化和功能适应。出生后肾脏的生长和成熟起着重要作用在发育程序性疾病中发挥重要作用。例如，出生时体重适度减少的儿童肾单位数量可能对代谢应激更敏感，例如高脂肪或高盐饮食、肥胖和缺氧或肾毒素引起的肾损伤。许多研究已经生成了胎儿和成人的单细胞数据集小鼠肾。然而，尽管产后侮辱在发育和进展中发挥着重要作用， CKD，即出生后肾脏的成熟尚未在分子水平上得到全面研究使用最先进的技术。由于缺乏这一关键发展时期的参考图集，限制了我们的研究能够研究小儿肾脏疾病模型中细胞状态的变化和多样性。肾小球疾病，包括肾病综合征，是儿童肾病的一个重要原因疾病。局灶性节段性肾小球硬化症 (FSGS) 是一种频率不断增加的诊断，经常复发且对治疗有抵抗力，并导致终末期肾病。继发性肾小球硬化和蛋白尿在影响儿童的常见非肾小球疾病（例如 CAKUT）中出现，并预示着进展为终末期肾衰竭的风险较高。目前，治疗选择有限这些疾病以及迫切需要更好地了解疾病机制。主要机制之一细胞对压力或损伤做出反应是利用染色质修饰机制来重新编程基因监管网络。我们开发了一种由于转移功能丧失而导致 FSGS 的新型小鼠模型相关蛋白 2 (Mta2)，NuRD 染色质重塑复合物的核心成分。这些突变体小鼠代表了一个很好的模型来研究肾单位禀赋的临床沉默缺陷和细胞分化的改变可在青春期晚期/成年早期表现为 FSGS。在本提案中，我们将生成一个集成的单细胞和空间转录组学小鼠图谱 (pKidCAP) 在产后时间点与主要生理和发育变化一致肾。我们将确定细胞多样性、空间定位、单细胞基因表达的进化健康和 FSGS 组织中发育中的产后肾脏的特征和染色质可及性状态。我们将应用 ChIP-seq 和表观基因组编辑来测试 Jun/AP-1 靶标去抑制的假设 Mta2 突变体会促进炎症、细胞衰老和脂毒性。将这些数据集与人类 pKidCAP 图谱（项目 1）将创建一个独特的参考面板来识别小鼠中发现的通路与儿童肾脏疾病高度相关的模型。