DEVELOPMENTAL ROLE OF THE GENE SIX2 IN KIDNEY NEPHRON ENDOWMENT

基因 SIX2 在肾单位发育中的作用

• 批准号: 8167760
• 负责人: SUWIT JACK SOMPONPUN
• 金额: $ 3.83万
• 依托单位: UNIVERSITY OF HAWAII AT MANOA
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-07-01 至 2011-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8167760
• 关键词: Adult; Chronic Kidney Failure; Computer Retrieval of Information on Scientific Projects Database; Development; Elderly; Endowment; Funding; Gene Expression; Genes; Genetic; Genetic Transcription; Goals; Grant; Growth; Homeobox Genes; Hypertension; Institution; Kidney; Kidney Diseases; Metanephric Diverticulum; Methodology; Morphogenesis; Mouse Strains; Mus; Mutant Strains Mice; Nephrons; Organogenesis; Phenotype; Physiological; Physiology; Production; RNA Interference; Research; Research Personnel; Resources; Role; Source; United States National Institutes of Health; Work; base; cardiovascular risk factor; design; insight; mouse genome; mouse model; nephrogenesis; research study; restoration

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Suboptimal kidney development resulting in an in-born deficit in nephron number can have lifelong consequences and is associated with a significant risk of cardiovascular-renal disease in later life, compared to those born with a typical nephron endowment. The long-term goal of the current proposal is to identify cellular mechanisms that drive successful nephron differentiation to achieve optimal nephron endowment. Specifically, we will explore the roles of the homeobox gene sine oculis 2 (six2), and determine whether six2 is involved in the establishment of nephron number during kidney organogenesis. Using an RNAi methodology, experiments are designed to determine whether inactivation of six2 transcription early in nephrogenesis results in a significant decrease in ureteric bud growth and, therefore, nephron number in an organotypic kidney explant culture. Further, using a mouse model of heritable renal hypoplasia that lacks sufficient expression of six2 during development (the Brachyrrhine (Br) mutant mouse), we will take a systematic approach to re-introduce six2 to kidney explants that are prepared from Br mice and determine if exogenous six2 stimulates branching of the ureteric buds leading to an enhanced nephron production. Additionally, we will incorporate a 250 kb Bac containing six2 gene into the Br mouse genome to attempt to increase nephron number and, thereby rescuing the defective renal phenotype associated with six2 deficiency. Following the rescue we will assess if renal physiological features are restored in the adult Br mouse. This proposal will take advantage of the Br mouse strain that displays haploinsufficiency of six2 gene expression and is the only working colony in the world. Similarly, we have fully characterized the physiology of the adult Br mouse that demonstrates hypertension and chronic renal failure facilitating our phenotypic rescue experiments. When considered together, these experiments will provide the fundamental insights that constitute the genetic basis of six2 in renal morphogenesis and specifically establish whether six2 directly determines nephron endowment in the developing kidney. Results from this study will also underscore the importance of six2 in possible nephron restoration approaches in the adult diseased kidney.

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目及 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 对于中心来说，它不一定是研究者的机构。 肾脏发育不理想会导致先天性肾单位数量不足 终生后果，并与心血管肾疾病的显着风险相关 与那些出生时具有典型肾单位禀赋的人相比，在以后的生活中。长期目标 当前提案的目的是确定驱动肾单位成功的细胞机制 分化以实现最佳肾单位禀赋。具体来说，我们将探讨角色 的同源框基因 sine oculis 2 (six2)，并确定 Six2 是否参与 肾器官发生过程中肾单位数量的建立。使用 RNAi 方法， 实验旨在确定 Six2 转录的早期失活是否 肾发生导致输尿管芽生长显着减少，因此肾单位 器官型肾外植体培养中的数量。此外，使用可遗传的小鼠模型 肾发育不全，在发育过程中缺乏足够的 Six2 表达（短鼻 （Br）突变小鼠），我们将采取系统的方法将 Six2 重新引入肾脏 从 Br 小鼠制备外植体并确定外源性 Six2 是否刺激 输尿管芽的分支导致肾单位生成增强。此外，我们 将把包含 Six2 基因的 250 kb Bac 整合到 Br 小鼠基因组中，以尝试 增加肾单位数量，从而挽救相关的肾表型缺陷 与六2缺乏。抢救后，我们将评估肾脏生理特征是否正常 在成年 Br 小鼠中恢复。该提案将利用 Br 小鼠品系， 显示出 Six2 基因表达的单倍体不足，并且是该群体中唯一的工作菌落 世界。同样，我们已经充分描述了成年 Br 小鼠的生理学特征 证明高血压和慢性肾功能衰竭有助于我们的表型拯救 实验。当综合考虑时，这些实验将提供基本的 构成肾形态发生中的 Six2 遗传基础的见解，特别是 确定 Six2 是否直接决定发育中肾脏的肾单位禀赋。 这项研究的结果还将强调 Six2 在可能的肾单位中的重要性 成人患病肾脏的恢复方法。