Ex Vivo Generation of Functional Kidney Tissues for Transplantation

用于移植的功能性肾组织的体外生成

• 批准号: 10053515
• 负责人: Jennifer A. Lewis
• 金额: $ 79.03万
• 依托单位: HARVARD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-01 至 2025-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10053515
• 关键词: 3-Dimensional; Address; Affect; Biomanufacturing; Biomedical Engineering; Blood Vessels; Chronic Kidney Failure; Coculture Techniques; Complex; Developmental Biology; Devices; Dialysis procedure; Disease model; Distal; Drainage procedure; Duct (organ) structure; Ductal Epithelial Cell; Electrolytes; End stage renal failure; Engineering; Epithelial; Epithelium; Exhibits; Extracellular Matrix; Filtration; Foundations; Generations; Goals; Heterogeneity; Homeostasis; Human; In Vitro; Individual; Kidney; Kidney Transplantation; Lead; Liquid substance; Methods; Micropuncture; Modeling; Morbidity - disease rate; Nephrons; Nutrient; Organ; Organ Donor; Organ Transplantation; Organogenesis; Organoids; Output; Oxygen; Patients; Physiological; Printing; Production; Protocols documentation; Renal Replacement Therapy; Renal Tissue; Renal function; Research; Structure; System; Technology; Therapeutic; Therapeutic Uses; Tissue Transplantation; Tissues; Transplantation; Tubular formation; Ureter; Validation; Vascularization; Venous; Xenobiotics; bioprinting; blood filter; capillary bed; density; experimental study; fluid flow; glomerular filtration; glomerular function; human pluripotent stem cell; human stem cells; in vivo; induced pluripotent stem cell; mortality; self assembly; shear stress; societal costs; stem cells; success; tissue repair; urinary; wasting

PROJECT SUMMARY In the U.S. alone, up to 26 million people have chronic kidney disease, over 460,000 people are on dialysis, and 100,000 people await kidney transplants with 3,000 new patients added monthly. Given the growing lack of transplantable organs, patients typically require renal replacement therapies that themselves lead to substantial morbidity and mortality. We posit that biomanufactured kidney tissues, and ultimately, organs may offer an important solution to this growing problem. Indeed, recent protocols in developmental biology are unlocking the potential for stem cells to undergo differentiation and self-assembly to form “mini-organs”, known as organoids. Kidney organoids exhibit remarkable tissue microarchitectures with high cellular density and heterogeneity akin to their in vivo counterparts. To bridge the gap from these kidney organoid building blocks (OBBs) to therapeutic organs, integrative approaches that combine bottom-up organoid assembly with top-down bioprinting are needed. While it is difficult, if not impossible, to imagine how either organoids or bioprinting alone would fully replicate the complex multiscale features required for kidney function – their combination could provide an enabling foundation for de novo organ manufacturing. To generate 3D functional kidney tissues ex vivo for potential transplantation, our highly collaborative research team will undertake two primary aims. In Specific Aim 1, we will create kidney organoids enhanced by multilineage induction that display functional differentiation of nephrons. We will produce iPSC-derived kidney organoids and subject them to fluid flow during their differentiation and maturation on an adherent extracellular matrix (ECM). Through multilineage induction, we will also induce collecting duct cells that self-assemble and structurally bridge other tubular nephron segments. We will evaluate the effects of mimicking kidney organogenesis on kidney organoid structure and function using microperfusion and micropuncture methods. In Specific Aim 2, we will create 3D functional kidney tissues composed of these optimized kidney OBBs with embedded macrochannels produced by bioprinting that serve as both vascular and urinary output conduits. We will first produce a densely cellular, tissue matrix composed of kidney OBBs that facilitates bioprinting of embedded macrochannels. We will then establish connections between the printed macrochannels embedded in this OBB-laden matrix and the self-assembled microvascular and collecting duct networks within individual OBBs. Finally, we will assess the glomerular filtration, tubular maturation, and primitive urinary production of these 3D kidney tissues. If successful, our proposed project will provide a foundational advance in kidney organ engineering for potential renal therapeutic applications.

项目总结 仅在美国，就有多达2600万人患有慢性肾脏疾病，超过46万人在接受透析，以及 10万人等待肾脏移植，每月新增3000名患者。鉴于日益缺乏的 对于可移植的器官，患者通常需要肾脏替代疗法，这些疗法本身就会导致实质性的 发病率和死亡率。我们假设，生物制造的肾脏组织，最终，器官可能提供 对这一日益严重的问题的重要解决方案。事实上，发育生物学的最新方案正在解锁 干细胞可能经历分化和自我组装，形成被称为类器官的“微型器官”。 肾脏有机化合物表现出显著的组织微结构，具有高细胞密度和类似的异质性 它们在体内的对应物。为了弥合这些肾脏器官积木(OBS)与治疗的差距 器官，将自下而上的有机体组装与自上而下的生物打印相结合的综合方法是 需要的。虽然很难想象，如果不是不可能的话，仅靠有机化合物或生物打印就可以完全 复制肾功能所需的复杂的多尺度特征-它们的组合可以提供 为新器官制造奠定基础。体外构建三维功能肾组织 对于潜在的移植，我们高度合作的研究团队将承担两个主要目标。以特定的目标 1，我们将创造通过多系诱导增强的肾脏器官，显示出功能分化 尼弗农。我们将生产IPSC衍生的肾脏有机化合物，并使它们在 在贴壁的细胞外基质(ECM)上分化和成熟。通过多血统的诱导，我们将 还可以诱导集合管细胞自组装并在结构上连接其他肾小管节段。我们 将评估模拟肾脏器官发生对肾脏器官结构和功能的影响 微灌注法和微穿刺法。在特定目标2中，我们将创建3D功能肾组织 由这些优化的肾脏OBB组成，其中嵌入了通过生物打印产生的大通道， 作为血管和尿液输出管道。我们将首先生产一个密集的细胞，组织基质，由 肾脏OBBS，便于嵌入的大通道的生物打印。然后，我们将建立连接 在嵌入OBB的基质中的印刷大通道和自组装的微血管之间 以及收集单个OBB内的风管网络。最后，我们将评估肾小球滤过、肾小管 这些3D肾组织的成熟和原始尿液产生。如果成功，我们提议的项目将 为潜在的肾脏治疗应用提供肾脏器官工程的基础性进展。