Induced pluripotent stem cells (iPSCs) developed from patients have enabled the investigation of disease mechanisms in the lab without dependence on animal models, which do not always mimic human disease conditions. As the cells differentiated from patient iPSCs also show the disease phenotype, they are very valuable in drug screening and testing. However, obtaining patient samples has issues of ethical concerns and accessibility. Further, the mutation from a specific patient may not represent the most prevalent disease variant.
Using genome editing, it is now possible to disrupt a gene function by introducing the mutation of interest and to correct a disease associated mutation. Therefore, to study a specific genetic disorder, the mutation of interest can be introduced in the target gene in a pluripotent stem cell derived from a normal donor. Once generated, the cell line can be differentiated to lineages to study disease pathogenesis or drug screening. Using the gene editing-based approach, we have successfully generated iPSC-based disease models for Pompe and ACADVL. In addition, using patient samples, iPSCs have been generated for the skeletal disorder Osteogenesis Imperfecta (OI). Currently, drug screening is being carried out using OI-iPSC-derived osteogenic cells.
Team
Vasanth Thamodaran
The medium used for culturing stem cells is prohibitively expensive with 80% of the cost attributed to cytokines (FLT-3L, SCF, TPO, IL-3, and IL-6). We aim to bring down the cost of cell-based therapy/gene therapy for hemoglobinopathies by in-house production of growth factors required for ex-vivo expansion of hematopoietic stem cells.
We have successfully expressed the growth factors IL3, IL6, SCF, and FLT3L in bacteria, refolded and purified. In addition, three growth factors – IL3, IL6, and SCF – were overexpressed in cytoplasm and purified. The growth factors were also validated using cell lines to check the growth-promoting activity. The validation showed that soluble cytoplasmic growth factors can promote cell proliferation severalfold.
Investigator: Vasanth Thamodaran