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.
We aim to generate pluripotent stem cell models for lysosomal disorders. Using the CRISPR-Cas gene editing tool, mutations that are widely prevalent in India will be introduced in pluripotent stem cells derived from normal donors. The cell lines carrying the disease associated mutation will then be differentiated to lineages affected by this disorder to study the disease pathogenesis. These disease models can also be used for testing biotherapeutics and drug screening.
The medium used for culturing these cells is prohibitively expensive and 80% of the cost is that of 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.
In collaboration with JSS Medical college (that provides a ready source of cord blood cells from which HSCs are produced), 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.
Investigator: Vasanth Thamodaran
JSS Medical college, Mysuru
The goal of the project is to generate pluripotent stem cell models for rare genetic disorders for disease modelling and testing biotherapeutics. Currently we are working on the following
- Lysosomal storage disorders (LSDs) prevalent in India by gene editing
- Skeletal myopathies by gene editing
- Osteogenesis Imperfecta (OI) from patient cells
Investigator: Vasanth Thamodoran
DBT – Institute for Stem Cell Science and Regenerative Medicine (DBT-InStem)
Christian Medical College (CMC), Vellore