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.

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 次优的肾脏发育导致出生时肾单位数量不足， 终身后果，并与心血管肾病的重大风险相关 在以后的生活中，与那些出生时具有典型肾单位禀赋的人相比。远景目标 目前的建议是确定细胞机制，驱动成功的肾单位， 分化以实现最佳肾单位禀赋。具体来说，我们将探讨 同源框基因sine oculis 2（six 2），并确定six 2是否参与了 肾器官发生过程中肾单位数目的建立。使用RNAi方法， 设计实验来确定six 2转录的失活是否在细胞的早期， 肾发生导致输尿管芽生长显著减少，因此，肾单位 器官型肾外植体培养物中的数量。此外，使用小鼠模型的遗传 肾发育不全，在发育过程中缺乏足够的six 2表达（短鼻型 (Br)突变小鼠），我们将采取系统的方法将six 2重新引入肾脏 从Br小鼠制备的外植体，并确定外源性six 2刺激 输尿管芽的分支导致肾单位产生增强。另外我们 将把一个250 kb的含有six 2基因的Bac基因整合到Br小鼠基因组中， 增加肾单位数量，从而挽救相关的缺陷性肾表型 6个缺陷抢救后，我们将评估肾脏生理特征是否 在成年Br小鼠中恢复。该提议将利用Br小鼠品系， 显示six 2基因表达的单倍不足，并且是该系统中唯一的工作菌落。 世界类似地，我们已经充分表征了成年Br小鼠的生理学特征， 证明高血压和慢性肾衰竭有助于我们的表型拯救 实验当考虑在一起，这些实验将提供基本的 这些见解构成了six 2在肾脏形态发生中的遗传基础， 确定six 2是否直接决定发育中肾脏的肾单位禀赋。 这项研究的结果也将强调six 2在可能的肾单位中的重要性。 成人患病肾脏的修复方法。