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

基本信息

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
项目类别：
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

项目摘要

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）：来自肾病患者的诱导多能干细胞（iPSC）在患者特异性疾病建模和免疫相容性组织替代治疗方面具有巨大潜力。申请人Benjamin Freedman博士在导师Joseph Bonventre博士的实验室中在这些领域进行了开创性研究。该申请的目标是进一步扩展候选人在这一新颖研究领域的专业知识和发现，使其成为全面、独立的研究项目。 Freedman 博士最近领导了针对多囊肾病 (PKD)（肾衰竭的主要原因）建立 iPSC 模型的研究。 PKD 是由多囊蛋白-1 (PC1)、PC2 和纤维囊蛋白/多导蛋白 (FPC) 突变引起的，它们在初级纤毛上相互作用。与健康或 ARPKD 患者的同等培养物相比，发现纤毛 PC2 减少是 ADPKD iPSC 以及后代上皮细胞和成肝细胞的共同特征。野生型 PC1 的过度表达挽救了 PC2 向纤毛的定位，这表明了一种可能的治疗方法。我们的实验室最近开发了用于将 iPSC 定向分化为表达肾谱系标记物的肾祖样细胞 (KPC) 的方案。利用现有和创新的 PKD iPSC 系，Freedman 博士将测试以下假设：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。为了在体内扩展这项工作，iPSC 衍生的 KPC 将被植入免疫缺陷小鼠体内以形成组织生长，并仔细检查其组织学和免疫组织化学证据，以支持肾脏分化和 PKD 特异性囊肿发生。这些实验将增进我们对 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，并扩展 Freedman 博士的技术库，包括基因组修饰、3D 培养和体内分化的新技能。 Freedman 博士将在该奖项下投入 100% 的时间进行研究，布莱根妇女医院将晋升他为教职。 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，并可使用 Bonventre 的所有设施，包括完成这些研究所需的所有设备。 Freedman 博士将得到哈佛大学 PKD 病理生理学和治疗领域三位著名专家的共同指导：Jing Zhou 博士、Friedhelm Hildebrandt 博士和 Theodore Steinman 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。 Freedman 博士将通过以下方式补充他的教育：1) 每周专门讨论干细胞、PKD 和肾脏生理学的会议和研讨会，2) 全国干细胞和肾脏病学会议，以及 3) 负责开展研究课程。预计他将在获奖期间每年撰写第一作者手稿，并在第三年年底前竞争独立研究奖。