Abstract
A generation of induced pluripotent stem cells (iPSC) by ectopic expression of OCT4, SOX2,
KLF4, and c-MYC has established promising opportunities for stem cell research, drug discovery,
and disease modeling. While this forced genetic expression represents an advantage, there will
always be an issue with genomic instability and transient pluripotency genes reactivation that
might preclude their clinical application. During the reprogramming process, a somatic cell must
undergo several epigenetic modifications to induce groups of genes capable of reactivating the
endogenous pluripotency core. Here, looking to increase the reprograming efficiency in somatic
cells, we evaluated the effect of epigenetic molecules 5-aza-20-deoxycytidine (5AZ) and valproic
acid (VPA) and two small molecules reported as reprogramming enhancers, CHIR99021 and A83-01,
on the expression of pluripotency genes and the methylation profile of the OCT4 promoter in a
human dermal fibroblasts cell strain. The addition of this cocktail to culture medium increased the
expression of OCT4, SOX2, and KLF4 expression by 2.1-fold, 8.5-fold, and 2-fold, respectively, with
respect to controls; concomitantly, a reduction in methylated CpG sites in OCT4 promoter region
was observed. The epigenetic cocktail also induced the expression of the metastasis-associated gene
S100A4. However, the epigenetic cocktail did not induce the morphological changes characteristic of
the reprogramming process. In summary, 5AZ, VPA, CHIR99021, and A83-01 induced the expression
of OCT4 and SOX2, two critical genes for iPSC. Future studies will allow us to precise the mechanisms
by which these compounds exert their reprogramming effects.
KLF4, and c-MYC has established promising opportunities for stem cell research, drug discovery,
and disease modeling. While this forced genetic expression represents an advantage, there will
always be an issue with genomic instability and transient pluripotency genes reactivation that
might preclude their clinical application. During the reprogramming process, a somatic cell must
undergo several epigenetic modifications to induce groups of genes capable of reactivating the
endogenous pluripotency core. Here, looking to increase the reprograming efficiency in somatic
cells, we evaluated the effect of epigenetic molecules 5-aza-20-deoxycytidine (5AZ) and valproic
acid (VPA) and two small molecules reported as reprogramming enhancers, CHIR99021 and A83-01,
on the expression of pluripotency genes and the methylation profile of the OCT4 promoter in a
human dermal fibroblasts cell strain. The addition of this cocktail to culture medium increased the
expression of OCT4, SOX2, and KLF4 expression by 2.1-fold, 8.5-fold, and 2-fold, respectively, with
respect to controls; concomitantly, a reduction in methylated CpG sites in OCT4 promoter region
was observed. The epigenetic cocktail also induced the expression of the metastasis-associated gene
S100A4. However, the epigenetic cocktail did not induce the morphological changes characteristic of
the reprogramming process. In summary, 5AZ, VPA, CHIR99021, and A83-01 induced the expression
of OCT4 and SOX2, two critical genes for iPSC. Future studies will allow us to precise the mechanisms
by which these compounds exert their reprogramming effects.
| Original language | English |
|---|---|
| Pages (from-to) | 1-12 |
| Number of pages | 12 |
| Journal | Molecules |
| Volume | 26 |
| Issue number | 1909 |
| DOIs | |
| Publication status | Published - 29 Mar 2021 |
