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）：来自肾脏疾病患者的诱导多能干细胞（IPSC）对于患者特异性疾病建模和免疫相容性组织替代疗法具有显着潜力。申请人本杰明·弗里德曼（Benjamin Freedman）博士在导师实验室的这些领域进行了开创性的研究，约瑟夫·邦文顿（Joseph Bonventre）博士。该应用程序的目标是将候选人的专业知识和发现进一步扩展到一个全面的独立研究计划中。 Freedman博士最近领导了建立IPSC多囊肾脏病（PKD）的研究，这是肾衰竭的主要原因。 PKD是由Polycystin-1（PC1），PC2和纤维细胞蛋白/聚糖素（FPC）中的突变引起的，这些突变在原发性纤毛上相互作用。与健康或ARPKD患者的同等培养物相比，发现纤毛PC2的减少是ADPKD IPSC和后代上皮细胞和肝母细胞中的常见特征。野生型PC1的过表达将PC2定位到纤毛中，这表明可能是一种治疗方法。最近在我们的实验室开发了方案，用于将IPSC的定向分化为表达肾脏谱系标记的肾脏祖细胞样细胞（KPC）。 Freedman博士利用现有和创新的PKD IPSC线，将检验以下假设：PKD疾病突变会导致2D培养中的去分化和细胞周期表型在3D培养中的异常囊肿发生。为了扩展体内这项工作，IPSC衍生的KPC将植入免疫缺陷的小鼠以形成组织生长，该组织将仔细检查，以确保肾脏分化和PKD特异性囊肿的组织学和免疫组织化学证据。这些实验将提高我们对PKD发病机理的理解，为未来的研究产生创新的细胞系和方法，并扩大Freedman博士的技术曲目，包括基因组修饰，3D培养和体内分化的新技能。 Freedman博士将100％的时间根据该奖项进行研究，而Brigham和妇女医院将把他晋升为教师职位。 Bonventre博士将每天继续指导Freedman博士，为他和他的研究助理提供办公室和板凳空间，并访问所有Bonventre设施，包括所有必要的设备来完成这些研究。 Freedman博士将在哈佛大学的PKD病理生理学和治疗方面的三名著名专家：Jing Zhou博士，Friedhelm Hildebrandt博士和Theodore Steinman博士共同征收。导师兼委托人将开会一次，以评估弗里德曼博士每六个月的进度。 Freedman博士将通过1）每周会议和专门针对干细胞，PKD和肾脏生理学的研讨会，2）国家干细胞和肾脏病学会议，以及3）负责任的研究课程进行研究。预计他将在奖励期间每年生产第一作者手稿，并在第三年结束之前获得独立研究奖的竞争。