INDUCIBLE PLURIPOTENT STEM CELLS AND KIDNEY REGENERATION

可诱导多能干细胞和肾脏再生

• 批准号: 7820510
• 负责人: JOSEPH VINCENT BONVENTRE
• 金额: $ 49.9万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-30 至 2011-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7820510
• 关键词: Address; Adult; Affect; Age-Years; Area; Back; Biomedical Engineering; Cardiovascular system; Cell Line; Cells; Chronic Kidney Failure; Development; Developmental Gene; Devices; Dialysis procedure; Effectiveness; Embryo; End stage renal failure; Engineering; Epithelial; Epithelial Cells; Ethics; Extracellular Matrix; Generations; Goals; Growth Factor; Health; Hepatocyte; Human; In Vitro; Injury; Intermediate Mesoderm; Kidney; Kidney Diseases; Kidney Transplantation; Life; Liver; Mesenchyme; Modality; Modeling; Morbidity - disease rate; Mus; Natural regeneration; Nephrons; Organ; Pancreas; Patients; Pluripotent Stem Cells; Population; Preparation; Process; Protocols documentation; Public Health; Regenerative Medicine; Renal Replacement Therapy; Renal function; Renal tubule structure; Reporter Genes; Series; Source; Staging; Stem Cell Research; Stem cells; System; Technology; Therapeutic; Time; Tissues; Toxic effect; Transplantation; Tubular formation; United States; Work; base; cell type; embryonic stem cell; gastrointestinal system; high throughput screening; human disease; human embryonic stem cell; human stem cells; in vitro Model; innovation; kidney cell; mortality; nephrogenesis; novel; progenitor; public health relevance; repaired; small molecule; small molecule libraries; stem; stem cell population; technique development; three dimensional structure; transcription factor; urologic

DESCRIPTION (provided by applicant): This application addresses broad Challenge Area (11) Regenerative Medicine and specific Challenge Topic, 11-DK-101: Promote regeneration and repair in the digestive system, liver, pancreas, kidneys, hematologic, and urologic system. Chronic kidney disease (CKD) is a serious global health problem associated with significant morbidity and mortality. Complete loss of kidney function results in end-stage renal disease (ESRD), and renal replacement therapies such as dialysis and transplantation are required to sustain life. Given the complications that are associated with these treatment modalities, cell-based strategies focused on replacing the cells that are lost in CKD and regenerating kidney tissue represent an innovative approach to treating CKD. Human inducible pluripotent stem (iPS) cells, a powerful and rapidly evolving technology in the field of regenerative medicine, could serve as a virtually unlimited patient-specific source of starting material for regenerating human kidney tissue. Currently, there are no existing protocols to generate renal epithelial cells from human iPS cells, embryonic stem (ES) cells, or any other cell type. The ultimate goal of the work proposed is to develop a stepwise protocol for the differentiation of human iPS cells into renal progenitor cells. A distinct population of renal progenitor cells could serve as a starting point for subsequent differentiation of glomerular and tubular epithelial cells. We will address this challenge with a two-step approach. In our first specific aim, we will use mouse reporter gene models in a high-throughput system to screen libraries of small molecules for candidates that can drive the differentiation of mouse ES cells towards a renal progenitor cell fate. The activation of the reporter genes odd-skipped related 1 (Osr1) and Six2 by small molecules will signify the successful differentiation of mouse ES cells into renal progenitor cells from the intermediate mesoderm, the precursor to all kidney tissue, and the metanephric mesenchyme, respectively. The identification of key small molecules would represent the first step in producing desired renal epithelial cells from ES cells. We will also explore the ability of other non-renal epithelial cells such as hepatocytes to be reprogrammed by small molecules into renal progenitor cells. In our second specific aim, we will establish a set of conditions whereby human iPS cells can be differentiated into renal progenitor cells in vitro. The effectiveness of small molecules, developmental transcription factors, exogenous growth factors, and extracellular matrix materials to induce the renal progenitor cell fate will be evaluated. We will also determine if adult mouse kidney proximal tubular cells can be reprogrammed into iPS cells and whether this reprogramming process involves reactivation of the kidney developmental genes Osr1 and Six2. Populations of human iPS cell-derived renal epithelial cells would be novel and important for multiple translational applications, including 1) incorporation into 3D structures to construct kidney tubules, as a first step to bioengineering patient-specific kidney tissue, 2) incorporation into renal assist devices to treat patients with severe forms of kidney injury, 3) development of in vitro models of patient-specific kidney diseases, and 4) the development of kidney-specific efficacy and toxicity screens for potential therapeutics. PUBLIC HEALTH RELEVANCE: Chronic kidney disease (CKD), the gradual and usually permanent loss of kidney function over time, is a significant worldwide public health problem affecting approximately 8-9% of the population over 40 years of age in the developed world. This is associated with very high cardiovascular mortality and is a great financial burden as patients progress to end stage renal disease and require dialysis or transplantation. Although recent advances in stem cell research have given hope to the use of human stem cells in the treatment of many human diseases, it is very important to understand how stem cells and kidney progenitors can be used to treat kidney disease. The purpose of this study is to develop strategies for transforming stem cells into cells of the kidney, with the ultimate goal of regenerating kidney tissue to treat patients with CKD.

描述（由申请人提供）：本申请涉及广泛的挑战领域（11）再生医学和特定的挑战主题，11-DK-101：促进消化系统、肝脏、胰腺、肾脏、血液和泌尿系统的再生和修复。慢性肾脏病（CKD）是一种严重的全球性健康问题，与显著的发病率和死亡率相关。肾功能的完全丧失导致终末期肾病（ESRD），需要肾脏替代治疗（如透析和移植）来维持生命。考虑到与这些治疗方式相关的并发症，基于细胞的策略专注于替换CKD中丢失的细胞并再生肾组织，这是治疗CKD的创新方法。人类诱导性多能干细胞（iPS）是再生医学领域中一种强大且快速发展的技术，可以作为再生人类肾组织的起始材料的几乎无限的患者特异性来源。目前，还没有从人iPS细胞、胚胎干（ES）细胞或任何其他细胞类型产生肾上皮细胞的现有方案。这项工作的最终目标是建立一个逐步的方案，将人iPS细胞分化为肾祖细胞。一个独特的群体的肾祖细胞可以作为一个起点，随后分化的肾小球和肾小管上皮细胞。我们将分两步应对这一挑战。在我们的第一个具体目标中，我们将在高通量系统中使用小鼠报告基因模型来筛选能够驱动小鼠ES细胞向肾祖细胞命运分化的候选者的小分子文库。小分子激活报告基因odd-skipped related 1（Osr 1）和Six 2将分别表示小鼠ES细胞从中间中胚层、所有肾组织的前体和后肾间充质成功分化为肾祖细胞。关键小分子的鉴定将代表从ES细胞产生所需肾上皮细胞的第一步。我们还将探索其他非肾上皮细胞（如肝细胞）被小分子重编程为肾祖细胞的能力。在我们的第二个具体目标中，我们将建立一组条件，使人iPS细胞可以在体外分化为肾祖细胞。将评价小分子、发育转录因子、外源性生长因子和细胞外基质材料诱导肾祖细胞命运的有效性。我们还将确定成年小鼠肾脏近端小管细胞是否可以重编程为iPS细胞，以及这种重编程过程是否涉及肾脏发育基因Osr 1和Six 2的重新激活。人iPS细胞衍生的肾上皮细胞群对于多种翻译应用将是新颖的和重要的，包括1）并入3D结构以构建肾小管，作为生物工程化患者特异性肾组织的第一步，2）并入肾辅助装置以治疗患有严重形式的肾损伤的患者，3）开发患者特异性肾病的体外模型，和4）开发潜在治疗剂的肾特异性功效和毒性筛选。 公共卫生相关性：慢性肾脏疾病（CKD）是一种随着时间推移逐渐且通常永久性丧失肾功能的疾病，是一种重要的全球性公共卫生问题，影响发达国家40岁以上人口的约8-9%。这与非常高的心血管死亡率相关，并且随着患者进展到终末期肾病并需要透析或移植，这是巨大的经济负担。尽管干细胞研究的最新进展为人类干细胞在许多人类疾病的治疗中的应用带来了希望，但了解干细胞和肾祖细胞如何用于治疗肾脏疾病是非常重要的。本研究的目的是开发将干细胞转化为肾脏细胞的策略，最终目标是再生肾脏组织以治疗CKD患者。