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）：急性肾损伤 (AKI) 和慢性肾病 (CKD) 是退伍军人群体的严重健康问题。尽管有足够的最佳药物治疗，但受伤后肾小管无法修复，肾功能下降往往会导致透析和显着的发病率和死亡率。日益增长的需求与有限的可移植器官供应之间日益扩大的差距现在成为阻碍肾移植扩展到所有患有终末期肾病的患者和退伍军人的主要限制障碍，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，以解码肾脏修复机制和影响组织再生的途径，为未来的临床干预修复提供机会我们解决了干细胞技术目前的局限性，以描述和定位三维 (3D) 组织中干细胞成熟的机制。为了定义这些基本参数，我们开发了非细胞 3D 支架并对其进行了表征。脱细胞啮齿动物肾脏，可以研究干细胞和祖细胞向后肾间充质的生长和分化，并最终形成成熟的肾谱系。来自我们小组以及其他人的证据表明，来自于的信号。我们的研究结果表明，3D 细胞外基质 (ECM) 支架以地理特异性方式诱导干细胞分化，E-钙粘蛋白以基质排列的小管为中心，具有生物活性。具体的已知具有介导肾器官发生的结构蛋白（例如胶原蛋白、层粘连蛋白）和生长因子（例如 VEGF、bFGF）的 ECM 我们将检验以下假设：来自 3D 肾 ECM 组织的信号是引导干细胞和肾祖细胞正确分化的先决条件。肾脏细胞的复杂性促使我们特别关注无细胞基质支架内的肾小管发生，并确定体外再生和组织形成的要求。将使用两个肾干细胞/祖细胞群：能够嵌入发育中的肾单位的早期肾源性成年肾祖细胞，以及分化形成早期后肾组织的多能重编程人类 iPSC 和胚胎干细胞群。 3D 支架和干细胞旨在阐明驱动分化的干细胞-基质相互作用的要素目标 1 将研究干/祖细胞对肾 ECM 修饰的反应在高通量系统中，使用小型 ECM 支架快速筛选有利于分化的条件。在此过程中，我们确定了必要的基质结合生长因子并测试了它们。肾基质成纤维细胞 ECM 重塑的作用将通过分析大型器官大小的支架中干/祖细胞生长的需求来评估可扩展性，并探索灌注生物反应器内的肾小管形成。该研究策略使用创新的基质技术来修改组织支架，并建立了全尺寸的肾脏支架，并确定了正常和患病 ECM 内干细胞分化和支架重新增殖的要求。我们的系统研究非常重要，因为它将解码参与肾单位重建的关键因素，这是肾小管修复和肾组织再生的关键后续步骤。