Patient-Specific Human iPSCs for Modeling TSC Pathophysiology and Therapeutic Discovery
Abstract
Tuberous sclerosis complex (TSC) patients show symptoms in many organs that affect a variety of cell types in the body. Recent technical advances in the stem cell field has made it possible to culture patient-specific skin cells and then induce them to become pluripotent stem cells (also known as iPSCs). Because of their pluripotent nature, these iPSCs can then give rise to many different cell types in the body (such as neurons, kidney, or heart cells) under different culturing conditions. We have made tremendous progress in generating iPSCs directly from cultured skin cells of two unrelated TSC individuals. These iPSCs could then be converted into neural crest cells, which are thought to be the cells of origin affected in patients with TSC. Why do we need human cellular models when mouse models for TSC are available? Mouse models have certainly helped in identifying pathways affected in TSC as well as in drug studies; however, they have limitations since they do not mimic the human condition. In particular, many of the neurological symptoms and the tumor growth in many different organs are not seen in mouse models of TSC. We believe that the TSC proteins may play specific role(s) in human cell types, and we propose to understand this specificity by studying cultured human cells from two TSC patients (one with TSC1 and the other with TSC2). Importantly, we use the revolutionary and exciting new CRISPR/Cas9 technique to make genetically matched identical cell line sets, with and without TSC1 or TSC2 expression, to directly examine what abnormalities/altered mechanisms may occur in these patient-derived cell lines when TSC proteins are gone. Once we have these genetically matched sets in place, they will be invaluable in performing drug screening studies with the goal of identifying chemical compounds that are more effective for treating TSC patients than what is currently available. Ultimately, the research that we propose here will set the stage for better understanding of the underlying molecular mechanisms that take place in the brains of TSC patients as well as other organs. Although the nature of this project is basic research, the stem cells generated will be of great potential for immediate drug screening and other advances that may happen in the future. The experience we gain from this project will then allow us to take a similar approach and generate stem cells from additional TSC patients, ultimately leading to better understanding of differences in disease severity in individuals with TSC. Our proposed work addresses Fiscal Year 2014 Tuberous Sclerosis Complex Research Program focus areas such as "developmental origin of TSC manifestations" under the category Personalization of Care, as well as "continued refinement of cell-based and preclinical models" that falls under Optimization of Treatments. Therefore, we believe that our proposed work will have a significant and direct impact on the treatment and management of TSC.
Document Details
- Document Type
- DoD Grant Award
- Publication Date
- Mar 29, 2016
- Source ID
- W81XWH1510414
Entities
People
- Vijaya Ramesh
Organizations
- Massachusetts General Hospital
- United States Army