Epigenetic repression of CHCHD2 enhances survival from single cell dissociation through attenuated Rho A kinase activity.
Jumee KimEun-Ji KwonYun-Jeong KimDayeon KimYoon-Ze ShinDayeon GilJung-Hyun KimHyoung Doo ShinLyoung Hyo KimMi-Ok LeeYoung-Hyun GoHyuk-Jin ChaPublished in: Cellular and molecular life sciences : CMLS (2024)
During in vitro culture, human pluripotent stem cells (hPSCs) often acquire survival advantages characterized by decreased susceptibility to mitochondrial cell death, known as "culture adaptation." This adaptation is associated with genetic and epigenetic abnormalities, including TP53 mutations, copy number variations, trisomy, and methylation changes. Understanding the molecular mechanisms underlying this acquired survival advantage is crucial for safe hPSC-based cell therapies. Through transcriptome and methylome analysis, we discovered that the epigenetic repression of CHCHD2, a mitochondrial protein, is a common occurrence during in vitro culture using enzymatic dissociation. We confirmed this finding through genetic perturbation and reconstitution experiments in normal human embryonic stem cells (hESCs). Loss of CHCHD2 expression conferred resistance to single cell dissociation-induced cell death, a common stress encountered during in vitro culture. Importantly, we found that the downregulation of CHCHD2 significantly attenuates the activity of Rho-associated protein kinase (ROCK), which is responsible for inducing single cell death in hESCs. This suggests that hESCs may survive routine enzyme-based cell dissociation by downregulating CHCHD2 and thereby attenuating ROCK activity. These findings provide insights into the mechanisms by which hPSCs acquire survival advantages and adapt to in vitro culture conditions.
Keyphrases
- single cell
- cell death
- copy number
- pluripotent stem cells
- dna methylation
- genome wide
- protein kinase
- rna seq
- gene expression
- endothelial cells
- mitochondrial dna
- high throughput
- electron transfer
- embryonic stem cells
- oxidative stress
- risk assessment
- cell proliferation
- poor prognosis
- cell therapy
- binding protein
- high glucose
- hydrogen peroxide
- diabetic rats
- stem cells
- smooth muscle
- single molecule
- drug induced
- stress induced
- mesenchymal stem cells
- tyrosine kinase