Extracellular Matrix Induction of Renal Stem and Progenitor Cell Development

肾干细胞和祖细胞发育的细胞外基质诱导

基本信息

批准号：
10200365
负责人：
JASON A WERTHEIM
金额：
--
依托单位：
SOUTHERN ARIZONA VA HEALTH CARE SYSTEM
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-10-01 至 2022-09-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10200365
关键词：
3-Dimensional Acute Renal Failure with Renal Papillary Necrosis Address Adult Affect American Binding Biological Markers Biomedical Engineering Bioreactors Bladder Bladder Dysfunction Cardiovascular system Cell Differentiation process Cell Maturation Cells Chronic Kidney Failure Clinical Coculture Techniques Collagen Communities Complex Controlled Environment Cues Development Diabetes Mellitus Dialysis procedure Differentiation and Growth Disease E-Cadherin Elderly Elements End stage renal failure Environment Evaluation Excision Expenditure Extracellular Matrix FGF2 gene Fibroblasts Financial Hardship Future General Population Genomic medicine Geography Goals Growth Growth Factor Health Hemodialysis High Prevalence Human Hypertension Immunosuppression In Vitro Individual Injury Intercellular Junctions Intervention Investigation Kidney Kidney Diseases Kidney Failure Kidney Transplantation Laboratories Laminin Ligands Living Donors Mediating Medical Medicare Mesenchyme Methods Modeling Modification Monitor Morbidity - disease rate Natural regeneration Nephrons Organ Organ Culture Techniques Organ Size Organ Transplantation Organogenesis Paracrine Communication Pathway interactions Patients Peptides Perfusion Peripheral Vascular Diseases Population Process Public Health Renal Tissue Renal function Renal tubule structure Rodent Role Services Signal Transduction Skin Stem Cell Development Structural Protein Structure System Technology Testing Therapeutic Uses Time Tissue Engineering Tissue Transplantation Tissues Transplantation Ursidae Family Vascular Endothelial Growth Factors Veterans Waiting Lists base cell type diabetic ulcer embryonic stem cell first-in-human health administration high throughput screening human pluripotent stem cell improved induced pluripotent stem cell injury and repair innovation mortality personalized medicine preservation prevent progenitor public health relevance reconstitution repaired response scaffold stem stem cell biology stem cell differentiation stem cell technology stem cells three dimensional cell culture tissue regeneration tissue support frame tool two-dimensional

项目摘要

DESCRIPTION (provided by applicant): Acute kidney injury (AKI) and chronic kidney disease (CKD) are critical health problems in the Veteran community. In the absence of sufficient tubule repair after injury and despite the best medical therapy, decreasing kidney function often leads to dialysis and significant morbidity and mortality. The growing gap between the increasing demand and limited supply of transplantable organs is now the chief limiting obstacle preventing extension of kidney transplantation to all patients and veterans with end-stage renal disease. Stem cell biology, and in particular induced pluripotent stem cell (iPSC) technology, provides new strategies via innovative advances in genomic and personalized medicine to decode the mechanisms of renal repair and pathways influencing tissue regeneration, to offer an opportunity for future clinical intervention to repair injured renal tubules or replace failed renal tissue. We address the current limitation in stem cel technology-to delineate and target mechanisms of stem cell maturation in three-dimensional (3D) tissue. To define these fundamental parameters we developed and characterized acellular 3D scaffolds from decellularized rodent kidneys, which allow study of growth and differentiation of stem and progenitor cells toward metanephric mesenchyme, and ultimately a mature renal lineage. Evidence from our group, as well as others, indicate that signals from 3D extracellular matrix (ECM) scaffolds induce stem cell maturation. Our findings show stem cell differentiation in a geographically-specific manner with maturation and expression of E-cadherin centered around matrix-lined tubules. Our 3D scaffolds are bioactive, consisting of natural tissue- specific ECM with structural proteins (e.g. collagens, laminins) and growth factors (e.g. VEGF, bFGF) known to mediate renal organogenesis. We will test the hypothesis that signals from 3D renal ECM tissue are a prerequisite to direct proper stem and renal progenitor cell differentiation to form nephron segments. The cellular complexity of the kidney prompts us to specifically focus on tubulogenesis within acellular matrix scaffolds and determine the requirements for regeneration and tissue formation in vitro. We will use two renal stem/progenitor populations: an early kidney-derived, adult renal progenitor cell capable of intercalating into developing nephrons, and a population of pluripotent reprogrammed human iPSCs and embryonic stem cells that differentiate to form early metanephric tissue. Our overall goal using 3D scaffolds and stem cells is to elucidate the elements of stem cell-matrix interactions that drive differentiation Aim 1 will investigate stem/progenitor cell response to modification of the renal ECM in a high-throughput system using small ECM scaffolds to rapidly screen conditions favoring differentiation. In this process we identify the requisite matrix-bound growth factors and test the role of ECM remodeling by renal stroma fibroblasts. Aim 2 will assess scalability by analyzing the requirements for stem/progenitor growth in larger-scale organ-sized scaffolds and explore tubule formation within a perfusion bioreactor. This system serves as a model for nephron development within a full-scale kidney scaffold and establishes the requirements for stem cell differentiation and scaffold repopulation within normal and diseased ECM. This investigative strategy uses innovative matrix technology to modify tissue scaffolds with fibroblasts and bioactive ligands to delineate mechanisms of differentiation in 3D. Our systematic investigation is highly significant because it will decode the critical factors involved in nephron reconstitutio, which are critical next steps in tubule repair and renal tissue regeneration.

描述（由申请人提供）：急性肾脏损伤（AKI）和慢性肾脏疾病（CKD）是老兵社区的关键健康问题。在受伤后没有足够的小管修复并希望进行最佳药物疗法的情况下，肾功能的降低通常会导致透析以及明显的发病率和死亡率。现在，需求不断增长和有限的可移植器官供应之间的差距日益增加，这是限制障碍的主要限制，以防止肾脏移植向所有患者和患有终末期肾脏疾病的退伍军人扩展。干细胞生物学，特别是诱导多能干细胞（IPSC）技术，通过基因组和个性化医学的创新进展提供新策略，以解码肾脏修复和途径的机制，并影响组织再生，以提供未来临床干预的机会受伤的肾管或替换失败的肾组织。我们解决了三维（3D）组织中干细胞成熟的茎技术的当前限制和干细胞成熟的目标机制。为了定义这些基本参数，我们开发并表征了脱细胞啮齿动物肾脏的细胞3D支架，从而可以研究茎和祖细胞对跨肾上腺素间充质的生长和分化，最终是成熟的肾谱系。来自我们组以及其他人的证据表明，来自3D细胞外基质（ECM）支架的信号会诱导干细胞成熟。我们的发现以地理特异性方式显示了干细胞分化，其成熟和表达的E-钙粘蛋白以基质衬里的管为中心。我们的3D支架具有生物活性，由天然组织特异性组成 ECM具有结构蛋白（例如胶原蛋白，层粘连蛋白）和生长因子（例如VEGF，BFGF）的ECM已知的肾脏器官。我们将测试以下假设：3D肾脏ECM组织的信号是指导正确的茎和肾脏祖细胞分化以形成肾单位段的先决条件。肾脏的细胞复杂性促使我们特别关注细胞基质支架内的结核病，并确定体外再生和组织形成的要求。我们将使用两个肾脏/祖细胞种群：一个早期肾脏衍生的成年肾脏祖细胞，能够插入发展为肾脏的肾脏，以及一个多能重编程的人IPSC和胚胎干细胞的种群，它们与众不同，它们与众不同，它们分化为早期的发态组织。我们使用3D支架和干细胞的总体目标是阐明驱动分化AIM 1的干细胞 - 基质相互作用的元素，将研究使用小型ECM脚手架在高通量系统中对肾脏ECM修饰的STEM/祖细胞响应，以快速筛选有利于分化的条件。在此过程中，我们确定了必要的矩阵结合增长因子，并测试 ECM重塑通过肾质基质成纤维细胞的作用。 AIM 2将通过分析大型有组织脚手架中的茎/祖细胞生长的要求来评估可伸缩性，并探索灌注生物反应器中的小管形成。该系统是全尺度肾脏支架内肾单位发育的模型，并确定了正常和解散的ECM内的干细胞分化和脚手架再现的要求。该调查策略使用创新的基质技术来修饰具有成纤维细胞和生物活性配体的组织支架，以描绘3D分化的机制。我们的系统投资非常重要，因为它将解释肾统一重构涉及的关键因素，这是Tuberepair和肾脏组织再生的关键下一步。