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: 10414819
• 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/10414819
• 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; 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; Persons; 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; directed differentiation; 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 cell derived tissues; 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.

项目概要 仅在美国，就有多达 2600 万人患有慢性肾病，超过 46 万人正在接受透析，并且 由于肾移植日益缺乏，目前有 10 万人等待肾移植，并且每月新增 3,000 名患者。 对于可移植器官，患者通常需要肾脏替代疗法，而肾脏替代疗法本身会导致大量的 我们认为生物制造的肾组织以及最终的器官可能会提供一种治疗方法。 事实上，发育生物学的最新协议正在解开这个日益严重的问题的重要解决方案。 干细胞具有分化和自组装形成“微型器官”（称为类器官）的潜力。 肾脏类器官表现出显着的组织微结构，具有高细胞密度和异质性 弥补这些肾脏类器官构件（OBB）与治疗之间的差距。 器官，将自下而上的类器官组装与自上而下的生物打印相结合的综合方法是 虽然很难（如果不是不可能的话）想象单独使用类器官或生物打印将如何完全实现。 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 为从头器官制造奠定基础 离体生成 3D 功能性肾组织。 在潜在的移植方面，我们高度合作的研究团队将实现两个主要目标。 1，我们将创建通过多谱系诱导增强的肾脏类器官，显示出功能分化 我们将生产 iPSC 衍生的肾脏类器官，并使其在其过程中处于流体流动状态。 通过多谱系诱导，我们将在贴壁细胞外基质（ECM）上分化和成熟。 还诱导集合管细胞自组装并在结构上桥接其他管状肾单位段。 将评估模仿肾脏器官发生对肾脏类器官结构和功能的影响 在具体目标 2 中，我们将创建 3D 功能性肾组织。 由这些优化的肾脏 OBB 组成，这些 OBB 具有通过生物打印产生的嵌入式大通道，可用于 作为血管和尿液输出导管，我们将首先产生由以下物质组成的致密细胞组织基质。 肾脏 OBB 有助于嵌入式宏通道的生物打印，然后我们将建立连接。 嵌入此 OBB 负载基质中的印刷大通道与自组装微血管之间 最后，我们将评估肾小球滤过、肾小管。 如果成功，我们提出的项目将实现这些 3D 肾组织的成熟和原始尿液生成。 为潜在的肾脏治疗应用提供了肾脏器官工程的基础性进展。