Epigenetic mechanisms of gene regulation in nephron progenitor cell proliferation and differentiation

肾单位祖细胞增殖和分化基因调控的表观遗传机制

基本信息

批准号：
10289761
负责人：
MICHAEL I RAUCHMAN
金额：
$ 42.22万
依托单位：
WASHINGTON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-07-01 至 2026-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10289761
关键词：
Address Adult Affect Area Back Binding Cell Differentiation process Cell Lineage Cells Childhood Chromatin Chromatin Remodeling Factor Chronic Kidney Failure Complement Complex Congenital Abnormality Cues Deacetylase Defect Development Disease Distal Embryo Endowment Enhancers Epigenetic Process Epithelial Cells Equilibrium Event Functional disorder Gene Expression Gene Expression Profile Gene Expression Regulation Genes Genetic Genome Genomics Goals Henle&apos s loop Human Human Development Human Genetics Hypertension Impairment Individual Injury Kidney Kidney Diseases Kidney Failure Knock-in Mouse Knowledge Lead Life Methods Modification Molecular Mus Natural regeneration Nephrons Nucleic Acid Regulatory Sequences Nucleosomes Organ Pattern Formation Physiological Play Polycomb Population Premature Birth Premature Infant Proteins Regulation Regulator Genes Regulatory Element Renal function Renal glomerular disease Research Role Signal Pathway Signal Transduction Sodium Chloride Sucrose Syndrome Testing Time Tissues Translating Tubular formation Undifferentiated WNT Signaling Pathway Water Work cell type conditional mutant end-stage organ failure epigenomics exhaustion genome-wide human disease induced pluripotent stem cell inhibitor/antagonist insight kidney cell mutant nephrogenesis nephron progenitor novel predictive modeling progenitor promoter renal epithelium repaired self-renewal stem cell growth stem cell proliferation stem cells transcription factor

项目摘要

The long-term goal of the proposed work is to determine mechanisms that control gene regulatory networks to generate a normal endowment of mature nephrons. Formation of the proper complement of nephrons requires a balance between self-renewal and differentiation of progenitor cells. Disruption of this balance leads to renal hypoplasia and chronic kidney disease. Soon after progenitor cells begin to differentiate, chromatin must undergo significant changes to set up gene expression patterns for formation of distinct cell types along the nephron. Defects in formation of specific renal epithelial cell types leads to glomerular disease and an inability of tubules to perform normal physiological functions, such as maintaining salt and water balance. A major mechanism by which cells respond to signals to direct gene expression to a particular fate or lineage is through the concerted action of chromatin remodeling complexes and tissue restricted transcription factors. These proteins act on regulatory regions in the genome to establish control of gene expression at the correct time and place. While individual factors that control gene expression in nephron progenitor cells have been defined, how these proteins cooperate to control gene expression in the developing kidney has not been explored. The identification and characterization of the regulatory elements where these factors act is also a major challenge in the field. We discovered that Sall1 plays a pivotal role in these developmental decisions by interacting with two distinct chromatin remodeling complexes, the Nucleosome Remodeling and Deacetylase (NuRD) complex and the Switch/Sucrose Non-Fermentable (SWI/SNF) complex. We propose that the integrated actions of Sall1, Six2, NuRD and SWI/SNF controls nephron progenitor cell gene expression to attain a dynamic balance between self-renewal and differentiation, and establishes the epigenetic modifications required for formation of specific cell lineages in the mature kidney. This novel paradigm will be tested in three Aims. In Aim 1, we will perform Hi-ChIP to identify enhancer-promoter contacts in uncommitted and induced progenitors, and define changes in mutant cells. Aim 2 will determine how genomic binding of Sall1, Six2, NuRD and SWI/SNF are affected in mutant nephron progenitor cells. Epigenomic editing will be used to demonstrate causal relationships between gene expression and binding of these factors at regulatory regions. In order to translate our findings to human development and disease, in Aim 3 we will test our findings in the mouse from Aims 1 and 2 in human iPS cell-derived nephron progenitor cells. These studies will illuminate mechanisms that underlie human genetic syndromes and sporadic birth defects, such as renal hypoplasia, which commonly cause childhood kidney failure. Our findings will also provide insight into how to reprogram kidney cells from a differentiated state back to a progenitor state, to promote regeneration of mature tubular epithelial cells in order to correct nephron deficits.

拟议工作的长期目标是确定控制基因调节网络的机制生成正常的成熟肾脏的赋。适当补充肾单位的形成需要在祖细胞的自我更新和分化之间保持平衡。这种平衡的破坏领先肾功能低迷和慢性肾脏疾病。祖细胞开始分化后不久，染色质必须发生重大变化以建立基因表达模式以形成沿着不同细胞类型的形成肾单位。特定肾上皮细胞类型的形成缺陷导致肾小球疾病和小管无法执行正常的生理功能，例如维持盐和水平衡。细胞对信号响应以将基因表达引导至特定命运或谱系通过染色质重塑复合物和组织限制转录的协同作用因素。这些蛋白质作用于基因组中的调节区域，以控制基因表达在正确的时间和地点。而控制基因表达在肾单位祖细胞中的个体因素具有被定义了，这些蛋白质如何合作控制发育中的肾脏中的基因表达探索。这些因素的作用也是一个调节元素的识别和表征也是一个该领域的主要挑战。我们发现SALL1通过与两个相互作用，在这些发展决策中起关键作用独特的染色质重塑络合物，核小体重塑和脱乙酰基酶（NURD）复合物和开关/蔗糖不可发酵（SWI/SNF）复合物。我们建议SALL1的综合行动， SIX2，NURD和SWI/SNF控制肾单位祖细胞基因表达以达到动态平衡在自我更新和分化之间，并建立形成所需的表观遗传修饰成熟肾脏中的特定细胞谱系。这个新颖的范式将以三个目标进行测试。在AIM 1中，我们将执行高芯片以识别未承诺和诱发祖细胞中的增强剂促进触点，并定义突变细胞的变化。 AIM 2将确定SALL1，SIX2，NURD和SWI/SNF的基因组结合如何在突变的肾单位祖细胞中受到影响。表观基因组编辑将用于证明因果关系基因表达与这些因素在调节区域的结合之间的关系。为了翻译我们对人类发展和疾病的发现，在AIM 3中，我们将从AIMS中测试鼠标中的发现1 在人IPS细胞衍生的肾单位祖细胞中2中。这些研究将阐明人类遗传综合征和零星出生的机制缺陷，例如肾功能低迷，通常会导致儿童肾衰竭。我们的发现也将提供有关如何从分化状态重新编程肾细胞回到祖细胞状态到祖细胞状态的洞察力促进成熟的管状上皮细胞的再生，以纠正肾脏缺陷。