DiMeN Doctoral Training Partnership: The role of mitochondrial dynamics and metabolism in preserving genomic integrity during reprogramming to induced pluripotent stem cells

Regenerative medicine aims to cure degenerative diseases by replacing the damaged or missing cells. Human pluripotent stem cells represent an essential pillar of regenerative medicine efforts as they provide a potentially unlimited source of progenitors or differentiated cells for use in cell replacement therapies and personalised medicine. Pluripotent stem cells can be obtained by reprogramming from adult somatic cells and we have recently developed a micro-carrier (MC)-based reprogramming platform that offers automatic, high-throughput, and large-scale production of induced pluripotent stem cells (iPSCs). Preservation of genomic integrity of cells during reprogramming is pivotal for their clinical applications since the genetic aberrations may affect the differentiation ability of iPSCs and/or functional characteristics of iPSC-derived differentiated cells. Indeed, a major concern in the field is that genetically variant iPSCs-derived cells may develop tumorigenic phenotypes upon transplantation in vivo.

Although genetically aberrant cells have been detected following reprogramming, it is not clear how the reprogramming process leads to genetically abnormal cells and whether reprogrammed cells may be compromised in their genomic stability during their subsequent expansion and differentiation. Mitochondrial function is now recognised as an important determinant of genomic stability. In particular, the necessity to re-programme mitochondria back to an ‘embryonic’ state if not controlled properly is likely to put metabolic stresses on cells and thus be an important factor in whether cells maintain their genetic integrity during re-programming.

Here, we will focus on determining the contribution of mitochondrial dynamics and metabolism in genome stability during the reprogramming process and whether metabolic stresses can be abrogated during reprogramming by careful manipulation of the culture conditions etc. Techniques will include somatic cell reprogramming using a high-throughput microcarrier-based platform, iPSC culture and the use of bioreactors, CRISPR/Cas9 genome editing, high-resolution and time-lapse imaging and general molecular biology and biochemistry techniques.