Growth Regulation and Nutrient Sensing in the Fetal Baboon Kidney

胎儿狒狒肾脏的生长调节和营养感应

• 批准号: 8609094
• 负责人: MARK J NIJLAND
• 金额: $ 18.68万
• 依托单位: UNIVERSITY OF TEXAS HLTH SCIENCE CENTER
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至 2015-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8609094
• 关键词: American; Amino Acids; Binding; Biological; Biological Assay; Brain; Cardiovascular Diseases; Cell Culture Techniques; Cell Cycle; Cell Cycle Progression; Cell Proliferation; Cellular biology; Child; Computer Simulation; Consumption; Culture Media; Developed Countries; Developing Countries; Development; Diabetes Mellitus; Diet; Dietary Intervention; Environment; Epigenetic Process; Epithelial; Epithelial Cell Proliferation; Epithelial Cells; Family member; Fetal Development; Fetal Growth; Fetal Growth Retardation; Fetal Kidney; Food; Fresh Tissue; Functional disorder; Funding; Genomics; Glucose; Growth; Health; Histones; Hour; Household; Human; Hunger; Incubated; Instruction; Intervention; Kidney; Leucine; Life; Link; Measures; Messenger RNA; Metabolic; Metabolism; Methods; Methylation; MicroRNAs; Molecular; Molecular Biology; NF-kappa B; Na(+)-K(+)-Exchanging ATPase; Nature; Nutrient; Nutritional; Ouabain; Outcome; Papio; Pathway interactions; Peptide Initiation Factors; Phenotype; Physiology; Predisposition; Pregnancy; Pregnant Women; Primates; Protein Biosynthesis; Proteins; Proteome; Proteomics; Public Health; Recording of previous events; Regulation; Relative (related person); Renal function; Risk; Signal Transduction; Sirolimus; Sodium; Stable Isotope Labeling; Supplementation; Suspension substance; Suspensions; Testing; Therapeutic Intervention; United States; Validation; Western Blotting; Woman; cell growth; cell transformation; detection of nutrient; epigenomics; experience; fetal; human FRAP1 protein; improved; inhibitor/antagonist; innovation; kidney cell; kidney cortex; mTOR Signaling Pathway; mTOR inhibition; maternal nutrient restriction; mother nutrition; nephrogenesis; nonhuman primate; nutrition; postnatal; pregnant; prevent; promoter; response; stressor; transcriptome sequencing; uptake

Periods of decreased nutrient availability represent the most common life-threatening stressors in nature. The mechanisms involved in defense against nutritional deficits are fundamental to survival and therefore highly redundant. One strategy relies on hypo-metabolism to facilitate survival in the face of decreased nutrient availability. In the United States at least 50 million Americans live In households experiencing food insecurity and hunger, exposing millions of women and children to decreased nutrient availability. Deficits in kidney development have been a central theme in studies demonstrating how decreased maternal nutrition impacts fetal development and postnatal cardio-renal function. Despite this history, the factors that mechanistically link reduced fetal nutrient availability and intrauterine growth restriction (IUGR) to poor renal development, i.e. renal genomic, epigenomic and proteomic responses to decreased nutrient availability that underlie renal dysfunction in fetal primates are essentially unknown. Fundamental to Project 3 is our central hypothesis that limited fetal nutrient availability associated with IUGR induces biological strategies acting to preserve development and promote survival in the face of decreased nutrition; in other words, the IUGR fetal environment represents a hypo-metabolic state that leads to deficits in kidney development. We will test this hypothesis by studying maternal nutrient restriction (MNR; 30% reduction in control [CTR] diet) and intervention through maternal dietary leucine supplementation (INT) in pregnant baboons. Four specific aims within P01 themes 2 (nutrient sensing), 3 (cell signalling) and 4 (function and epigenetics) examine the impact of MNR and INT on proximal tubule epithelial (PTE) cell nutrient sensing pathways (SAl), AMPK and mTOR signalling in PTE cell proliferation (SA2), epigenetic regulation of PTE cell growth (SA3), and PTE cell sodium transport function (SA4). Our approach utilizes Western blot, primary/ transformed cell culture, RNA Seq, miRNA Seq, proteomics (dimethylation/SILAC) and targeted promoter methylation. The significance lies in (1) studying maternal under nutrition, a serious public health problem, (2) exploring molecular mechanisms of non-human primate kidney development, and (3) improving understanding of the causes underiying complications of IUGR. Innovation lies in the multifaceted use of the non-human primate in integrating physiology, cell biology, proteomics, genomics, epigenomics, molecular biology and in silico methods to understand renal development and renal phenotype, creating the potential to devise therapeutic interventions to prevent and/or recuperate the impact of IUGR on kidney development and function.

营养素可用性下降的时期是自然界中最常见的危及生命的压力源。 参与防御营养缺乏的机制是生存的基础，因此 高度冗余。一种策略依赖于低代谢，以促进生存，在面对减少 养分有效性在美国，至少有5000万美国人生活在 不安全和饥饿加剧，使数百万妇女和儿童面临营养供应减少的风险。赤字 肾脏发育一直是研究的中心主题，这些研究表明， 影响胎儿发育和出生后的心肾功能。尽管有这样的历史， 将胎儿营养素利用率降低和宫内生长受限（IUGR）与肾功能不全 发育，即肾脏基因组、表观基因组和蛋白质组对营养物质可用性降低的反应， 灵长类胎儿肾功能不全的原因基本上是未知的。项目3的基础是我们的核心 与IUGR相关的胎儿营养有限的假设诱导生物学策略， 在营养下降的情况下保持发育并促进生存;换句话说，IUGR胎儿 环境代表导致肾脏发育缺陷的低代谢状态。我们将测试这个 通过研究母体营养限制（MNR;对照[CTR]饮食减少30%）假设， 在怀孕狒狒中通过母体饮食补充亮氨酸（INT）进行干预。四个具体 P01主题2（营养感测）、3（细胞信号传导）和4（功能和表观遗传学）中的目标研究了 MNR和INT对近曲小管上皮（PTE）细胞营养感应途径（SA 1）、AMPK和 PTE细胞增殖中的mTOR信号传导（SA 2）、PTE细胞生长的表观遗传调节（SA 3）和PTE细胞增殖中的mTOR信号传导（SA 4）。 钠转运功能（SA 4）。我们的方法利用蛋白质印迹，原代/转化细胞培养，RNA Seq、miRNA Seq、蛋白质组学（二甲基化/SILAC）和靶向启动子甲基化。意义 在于（1）研究母亲营养不良这一严重的公共卫生问题，（2）探索分子生物学， 非人灵长类动物肾脏发育的机制，以及（3）提高对原因的理解 胎儿宫内发育迟缓的并发症。创新在于对非人类灵长类动物的多方面利用， 整合生理学、细胞生物学、蛋白质组学、基因组学、表观基因组学、分子生物学和计算机模拟 了解肾脏发育和肾脏表型的方法，创造了设计治疗方法的潜力， 干预措施，以预防和/或恢复IUGR对肾脏发育和功能的影响。