Differentiation of Human iPS Cells into Kidney Proximal Tubular Cells

人 iPS 细胞分化为肾近端小管细胞

• 批准号: 7753366
• 负责人: Albert Lam
• 金额: $ 5.53万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-03-14 至 2012-03-13
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7753366
• 关键词: Acute; Acute Renal Failure with Renal Papillary Necrosis; Adult; Affect; Biomedical Engineering; Calcitriol; Candidate Disease Gene; Cell Line; Cells; Chronic Kidney Failure; Development; Developmental Gene; Devices; Dialysis procedure; Disease; Embryo; Embryonic Development; End stage renal failure; Engineering; Epithelial Cells; Ethics; Extracellular Matrix; Gene Expression; Gene Expression Profiling; Genes; Goals; Growth Factor; Human; Hypoxia; Injury; Kidney; Kidney Diseases; Kidney Transplantation; Molecular Profiling; Morbidity - disease rate; Multiple Myeloma; Mus; Natural regeneration; Nephrons; Organ; Patients; Pattern; Pharmacologic Substance; Physiology; Pluripotent Stem Cells; Polycystic Kidney Diseases; Population; Preparation; Process; Proximal Kidney Tubules; Public Health; Regenerative Medicine; Renal Cell Carcinoma; Renal function; Renal tubular acidosis; Renal tubule structure; Research; Role; Stem Cell Research; Stem cells; System; Testing; Time; Tissue Sample; Tissues; Toxic effect; Tubular formation; United States; Vitamin D; Water; Work; blastomere structure; human disease; human embryonic stem cell; human stem cells; in vitro Model; injured; interest; kidney cell; kidney epithelial cell; laser capture microdissection; mortality; nephrogenesis; novel; research study; solute; stem; stem cell population; technique development; three dimensional structure; transcription factor

DESCRIPTION (provided by applicant): The overarching goal of the work proposed is to understand the mechanisms of kidney development and regeneration with the long-term goal of establishing therapies for the treatment of acute and chronic kidney disease. Specifically, we will develop strategies to differentiate human inducible pluripotent stem (iPS) cells into proximal tubular epithelial cells. In our first specific aim, the goal is to understand gene expression profiles important to proximal tubule formation. We will approach this using two different experimental systems: 1) normal differentiation of proximal tubules during embryonic development, and 2) regeneration of proximal tubules following injury to the adult kidney. In both of these systems, proximal tubular cells will be isolated at key time points in each of these processes using laser capture microdissection (LCM). Using the collected tissue samples, gene expression profiling will be carried out using gene microarrays and real-time PCR. These experiments will allow us to select candidate genes that are important to inducing or regulating the proximal tubular cell fate. In the second specific aim, we will determine a set of conditions by which human iPS cells can be differentiated into proximal tubular cells. We will investigate the ability of our selected candidate genes and other genes known to be critical for normal kidney development to direct iPS cells toward the proximal tubular cell fate. We will also investigate the role of exogenous growth factors and extracellular matrix preparations in promoting the proximal tubular cell fate. Populations of human iPS cell-derived proximal tubular cells would be novel and important for incorporation into 3D structures to construct kidney tubules, as a first step to bioengineering patient-specific kidney tissue; incorporation into renal assist devices to treat patients with severe forms of kidney injury; development of in vitro models of patient-specific kidney diseases; and the development of kidney-specific toxicity screens for pharmaceuticals. Relevance: Chronic kidney disease (CKD), the gradual and usually permanent loss of kidney function over time, is a significant worldwide public health problem. Although recent advances in stem cell research have given hope to the use of human stem cells in the treatment of many human diseases, more research is needed to understand how stem cells can be used to treat kidney disease. The purpose of this study is to develop strategies for transforming stem cells into functioning kidney cells, with the ultimate goal of regenerating kidney tissue to treat patients with CKD.

描述（由申请人提供）：提出的工作的总体目标是了解肾脏发育和再生的机制，其长期目标是建立治疗急性和慢性肾脏疾病的疗法。具体而言，我们将制定将人诱导多能茎（IPS）细胞区分为近端管状上皮细胞的策略。在我们的第一个特定目标中，目标是了解对近端小管形成重要的基因表达谱。我们将使用两种不同的实验系统对此进行处理：1）胚胎发育过程中近端小管的正常分化，以及2）成年肾脏受伤后近端小管的再生。在这两个系统中，使用激光捕获微分解（LCM）在每个过程中的关键时间点将近端管状细胞分离。使用收集的组织样品，将使用基因微阵列和实时PCR进行基因表达分析。这些实验将使我们能够选择对于诱导或调节近端管细胞命运至关重要的候选基因。在第二个特定目的中，我们将确定一组可以将人IPS细胞区分为近端管状细胞的条件。我们将研究所选候选基因和其他已知对正常肾脏发育至关重要的能力，将IPS细胞引导到近端管状细胞命运。我们还将研究外源生长因子和细胞外基质制剂在促进近端管细胞命运中的作用。人IPS细胞衍生的近端管状细胞的群体对于掺入3D结构以构建肾小管的近端细胞将是新颖而重要的，这是生物工程特定于患者特异性肾脏组织的第一步。纳入肾脏辅助装置，以治疗严重形式的肾脏损伤患者；开发患者特异性肾脏疾病的体外模型；以及针对药物的肾脏特异性毒性筛查的开发。 相关性：慢性肾脏疾病（CKD）是随着时间的推移肾脏功能的逐渐且通常永久性损失，是全球范围内的重大公共卫生问题。尽管干细胞研究的最新进展使人们希望使用人类干细胞治疗许多人类疾病，但需要更多的研究来了解如何使用干细胞来治疗肾脏疾病。 这项研究的目的是制定将干细胞转化为功能性肾细胞的策略，其最终目的是再生肾脏组织以治疗CKD患者。