A 3D microperfusion model of autosomal dominant polycystic kidney disease

常染色体显性多囊肾病的 3D 微灌注模型

• 批准号: 8779922
• 负责人: Erica Palma Kimmerling
• 金额: $ 4.27万
• 依托单位: TUFTS UNIVERSITY MEDFORD
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-09-01 至 2016-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8779922
• 关键词: Accounting; Adenylate Cyclase; Affect; Autosomal Dominant Polycystic Kidney; Bilateral; Biocompatible Materials; Bioreactors; Cell Count; Cell Proliferation; Cell Survival; Cell-Matrix Junction; Cells; Cellular biology; Cilia; Cisplatin; Clinical; Collagen Type I; Cues; Culture Media; Custom; Cyst; Development; Dialysis procedure; Disease; Disease Outcome; Disease model; Doxycycline; Dyes; Elements; End stage renal failure; Engineering; Environment; Epithelial; Epithelial Cells; Gene Mutation; Goals; Growth; Growth Factor; Hereditary Disease; Histology; Human; Hypertrophy; Image; In Vitro; Injury; Interleukin-10; Ischemia; Kidney; Kidney Failure; Kidney Transplantation; Laboratories; Measures; Mechanics; Methodology; Methods; Microfluidic Microchips; Microfluidics; Modeling; Molecular Biology; Mutation; Nephrotoxic; Normal tissue morphology; Outcome; Pathogenesis; Pathway interactions; Patients; Perfusion; Phenotype; Polycystic Kidney Diseases; Porosity; Process; Renal Replacement Therapy; Renal dialysis; Renal tubule structure; Research; STAT6 gene; Scaffolding Protein; Scientist; Severity of illness; Silk; Simulate; Sodium Azide; Source; Structure; System; Testing; Tissue Engineering; Tissue Model; Tissues; Training; Transplantation; United States; base; cell growth; cytokine; design; experience; fluid flow; human FRAP1 protein; injury and repair; kidney epithelial cell; kidney repair; macrophage; matrigel; mimetics; monolayer; novel strategies; polycystic kidney disease 1 protein; public health relevance; repaired; response; scaffold; shear stress; simulation; skills; three-dimensional modeling; tissue repair

DESCRIPTION (provided by applicant): Autosomal dominant polycystic kidney disease (ADPKD) is a monogenic disorder that causes the development of bilateral focal cysts which ultimately result in renal failure and the need for renal replacement therapy such as dialysis or transplantation. Considering the disease affects over 600,000 people in the United States patients with ADPKD account for approximately 4% of all renal replacement therapy. Although the disease is associated with a mutation of either PKD1 (85% of cases) or PKD2 (15% of cases), there is a high level of variability between patients with respect to onset of cyst formation and disease severity. Due to a limited understanding of the disease pathogenesis there are no specific treatments for ADPKD and there is currently a lack of in vitro tissue models capable of elucidating the mechanisms behind cyst development. The goal of this project is to develop a 3D microperfusion model of ADPKD as a completely novel approach for investigating cytogenesis. The proposed methodology is a unique combination of tissue engineering and microfluidics which enables studies of cyst formation in response to mechanosensory cues, such as fluid flow, that is unattainable in current approaches. Fluid flow within this system can be used to evaluate the response of the tissue to changes in flow associated with renal injury and to introduce conditions mimicking renal repair. To achieve these goals a custom 3D perfusion system consisting of a microscale channel in a porous silk protein scaffold will be developed to provide the appropriate cell environment (aim 1a). A 3D in vitro human kidney tubule ADPKD disease model developed to have a controllable knockdown of PKD1 will be incorporated into the perfusion system for a comparison of normal and diseased tissues under perfusion and static conditions (aim 1b). The response of the normal and diseased tissues to injury based changes in fluid flow and subsequent repair stimulation will be characterized (aim 2). It is hypothesized that the forces mimicking injury will ultimately result in increased cell proliferation and cyst formation in the ADPKD model as the result of aberrant activation of affiliated pathways such as mTOR and STAT6. An increased understanding of the cellular pathways and external forces associated with cyst formation will ultimately assist in the development of targeted treatments for the disease. The structure of this proposal requires concurrent training in a diverse skill set including cell and molecular biology, biomaterials desig and engineering and bioreactor design and implementation and imaging. The diversity of research within the Kaplan lab provides the appropriate environment for pursuing the above proposal and desired training.

描述（由申请人提供）：常染色体显性多囊肾病（ADPKD）是一种单基因疾病，会导致双侧局灶性囊肿的发展，最终导致肾功能衰竭，并需要进行肾脏替代治疗，例如透析或移植。考虑到该疾病影响了美国超过 600,000 人，ADPKD 患者约占所有肾脏替代治疗患者的 4%。尽管该疾病与 PKD1（85% 的病例）或 PKD2（15% 的病例）突变有关，但患者之间在囊肿形成和疾病严重程度方面存在很大差异。由于对该疾病发病机制的了解有限，目前尚无针对 ADPKD 的特异性治疗方法，并且目前缺乏能够阐明囊肿发育背后机制的体外组织模型。该项目的目标是开发 ADPKD 的 3D 微灌注模型，作为研究细胞发生的全新方法。所提出的方法是组织工程和微流体的独特组合，使得能够研究响应机械感觉线索（例如流体流动）的囊肿形成，这在当前方法中是无法实现的。该系统内的流体流动可用于评估组织对与肾损伤相关的流动变化的反应，并引入模拟肾脏修复的条件。为了实现这些目标，将开发由多孔丝蛋白支架中的微通道组成的定制 3D 灌注系统，以提供适当的细胞环境（目标 1a）。为实现 PKD1 可控敲低而开发的 3D 体外人肾小管 ADPKD 疾病模型将被纳入灌注系统中，以比较灌注和静态条件下的正常组织和患病组织（目标 1b）。将表征正常组织和患病组织对基于损伤的液体流动变化和随后的修复刺激的反应（目标 2）。据推测，由于 mTOR 和 STAT6 等附属途径的异常激活，模拟损伤的力最终将导致 ADPKD 模型中细胞增殖和囊肿形成增加。加深对与囊肿形成相关的细胞途径和外力的了解将最终有助于开发针对该疾病的靶向治疗方法。该提案的结构需要同时进行多种技能的培训，包括细胞和分子生物学、生物材料设计和工程以及生物反应器设计和实施以及成像。卡普兰实验室内研究的多样性为实现上述建议和所需的培训提供了适当的环境。