Modeling Polycystic Kidney Disease Using Human Induced Pluripotent Stem Cells

使用人类诱导多能干细胞模拟多囊肾病

• 批准号: 8754901
• 负责人: Benjamin Solomon Freedman
• 金额: $ 15.8万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 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 2015-07-01
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8754901
• 关键词: Adult; Anatomy; Animal Model; Animals; Apoptosis; Architecture; Area; Autosomal Dominant Polycystic Kidney; Autosomal Recessive Polycystic Kidney; Award; Biological Assay; Blood Circulation; Cell Cycle; Cell Cycle Arrest; Cell Line; Cell model; Cells; Cilia; Clinical Trials; Complement; Complex; Cues; Cyst; Cystic kidney; Data; Defect; Disease; Disease model; Education; Epithelial; Epithelial Cells; Epithelium; Equipment; Exhibits; Faculty; Fibrosis; Functional disorder; Genes; Genome; Goals; Growth; Heterozygote; Hospitals; Human; Immunodeficient Mouse; Implant; In Vitro; Kidney; Kidney Diseases; Kidney Failure; Laboratories; Life; Manuscripts; Membrane; Mentors; Methodology; Modeling; Modification; Molecular; Mus; Mutation; Nephrology; Organelles; Organoids; Pathogenesis; Patients; Phenocopy; Phenotype; Physiology; Polycystic Kidney Diseases; Positioning Attribute; Pre-Clinical Model; Process; Proprotein Convertase 2; Protocols documentation; Replacement Therapy; Research; Resources; Sensory; Signal Transduction; Somatic Cell; Source; Stem cells; System; Teratoma; Testing; Therapeutic; Time; Tissues; Tube; Woman; Work; advanced disease; base; cell dedifferentiation; cell growth; cell type; cohort; disease characteristic; disease phenotype; embryonic stem cell; human disease; in vivo; induced pluripotent stem cell; innovation; insight; kidney cell; knockout animal; loss of function mutation; meetings; novel; overexpression; patient population; polycystic kidney disease 1 protein; progenitor; programs; public health relevance; repaired; research study; responsible research conduct; scaffold; self-renewal; skills; symposium; tool

DESCRIPTION (provided by applicant): Induced pluripotent stem cells (iPSCs) from patients with kidney disease have significant potential for patient-specific disease modeling and immunocompatible tissue replacement therapy. The applicant, Dr. Benjamin Freedman, has performed pioneering studies in these areas in the laboratory of the mentor, Dr. Joseph Bonventre. This application's goal is to further expand the candidate's expertise and findings in this novel research area into a well-rounded, independent research program. Dr. Freedman recently led research establishing iPSC models for polycystic kidney disease (PKD), a leading cause of kidney failure. PKD is caused by mutations in polycystin-1 (PC1), PC2, and fibrocystin/polyductin (FPC), which interact at the primary cilium. Reduced ciliary PC2 was found to be a common feature in ADPKD iPSCs and descendant epithelial cells and hepatoblasts, when compared to equivalent cultures from healthy or ARPKD patients. Overexpression of wild-type PC1 rescued PC2 localization to cilia, suggesting a possible therapeutic approach. Protocols have recently been developed in our laboratory for directed differentiation of iPSCs into kidney progenitor-like cells (KPCs) expressing markers of the renal lineage. Utilizing existing and innovative PKD iPSC lines, Dr. Freedman will test the hypothesis that PKD disease mutations result in dedifferentiation and cell cycle phenotypes in 2D culture and aberrant cystogenesis in 3D culture. To extend this work in vivo, iPSC-derived KPCs will be implanted into immunodeficient mice to form tissue growths, which will be carefully examined for histological and immunohistochemical evidence supporting kidney differentiation and PKD-specific cystogenesis. These experiments will advance our understanding of PKD pathogenesis, produce innovative cell lines and methodologies for future research, and expand Dr. Freedman's technical repertoire to include new skills of genome modification, 3D culture, and in vivo differentiation. Dr. Freedman will devote 100 % of his time to research under this award and Brigham and Women's Hospital will promote him to a faculty position. Dr. Bonventre will continue to mentor Dr. Freedman on a daily basis, providing office and bench space to him and his research assistants and access to all of the Bonventre facilities including all the necessary equipment to complete these studies. Dr. Freedman will be co-mentored by three renowned experts in PKD pathophysiology and treatment at Harvard: Dr. Jing Zhou, Dr. Friedhelm Hildebrandt, and Dr. Theodore Steinman. The mentor and co-mentors will meet to evaluate Dr. Freedman's progress every six months. Dr. Freedman will supplement his education with 1) weekly meetings and seminars devoted to stem cells, PKD, and kidney physiology, 2) national stem cell and nephrology conferences, and 3) responsible conduct of research courses. He is expected to produce first author manuscripts on an annual basis during the award period and will be competitive for independent research awards by the end of the third year.

描述（由申请人提供）：来自肾脏疾病患者的诱导多能干细胞（iPSC）具有用于患者特异性疾病建模和免疫相容性组织替代治疗的显著潜力。申请人Benjamin Freedman博士在导师Joseph Bonventre博士的实验室中进行了这些领域的开创性研究。该应用程序的目标是进一步扩大候选人在这个新的研究领域的专业知识和发现，成为一个全面的，独立的研究计划。Freedman博士最近领导了一项研究，为多囊肾病（PKD）建立iPSC模型，这是肾衰竭的主要原因。PKD是由多囊蛋白-1（PC 1）、PC 2和纤维囊蛋白/多导管蛋白（FPC）的突变引起的，它们在初级纤毛处相互作用。当与来自健康或ARPKD患者的等同培养物相比时，发现减少的纤毛PC 2是ADPKD iPSC和后代上皮细胞和成肝细胞中的共同特征。野生型PC 1的过表达拯救了PC 2定位于纤毛，这表明了一种可能的治疗方法。最近在我们的实验室中开发了用于将iPSC定向分化为表达肾谱系标志物的肾祖细胞样细胞（KPC）的方案。利用现有的和创新的PKD iPSC细胞系，Freedman博士将测试PKD疾病突变导致2D培养中的去分化和细胞周期表型以及3D培养中的异常囊肿发生的假设。为了在体内扩展这项工作，将iPSC衍生的KPC植入免疫缺陷小鼠中以形成组织生长，将仔细检查支持肾分化和PKD特异性囊肿形成的组织学和免疫组织化学证据。这些实验将促进我们对PKD发病机制的理解，为未来的研究产生创新的细胞系和方法，并扩大Freedman博士的技术储备，包括基因组修饰，3D培养和体内分化的新技能。弗里德曼博士将致力于100%的时间在这个奖项下的研究和布里格姆妇女医院将促进他的教师职位。Bonventre博士将继续每天指导Freedman博士，为他和他的研究助理提供办公室和工作台空间，并使用Bonventre的所有设施，包括完成这些研究所需的所有设备。Freedman博士将由哈佛三位著名的PKD病理生理学和治疗专家共同指导：Jing Zhou博士，Friedhelm Hildebrandt博士和西奥多斯坦曼博士。导师和共同导师将每六个月开会评估弗里德曼博士的进展。弗里德曼将补充他的教育与1）每周会议和研讨会，致力于干细胞，PKD和肾脏生理学，2）国家干细胞和肾脏学会议，和3）负责任的研究课程的行为。预计他将在获奖期间每年产生第一作者手稿，并将在第三年年底前竞争独立研究奖。