Comparing the Effects of Small Molecules Bix-01294, Bay K8644, Rg-108 and Valproic Acid, and Their Different Combinations on Induction of Pluripotency Marker-Genes By Oct4 in the Mouse Brain

Objective: every cell type is characterized by a specific transcriptional profile togetherwith a unique epigenetic landscape. reprogramming factors such as oct4, klf4, sox2and c-myc enable somatic cells to change their transcriptional profile and convert them topluripotent cells. small molecules such as bix-01294, bay k8644, rg-108 and valproicacid (vpa) are reported as effective molecules for enhancing induction of pluripotencyin vitro, however, their effects during in vivo reprogramming are addressed in this experimentalstudy.materials and methods: in this experimental study, oct4 expressing lentiviral particlesand small molecules bix-01294, bay k8644 and rg-108 were injected into the right ventricleof mice brain and vpa was systematically administered as oral gavages. animalstreated with different combinations of small molecules for 7 or 14 days in concomitant withoct4 exogenous expression were compared for expression of pluripotency markers. totalrna was isolated from the rims of the injected ventricle and quantitative polymerase chainreaction (pcr) was performed to evaluate the expression of endogenous oct4, nanog,c-myc, klf4 and sox2 as pluripotency markers, and pax6 and sox1 as neural stem cell(nsc) markers.results: results showed that oct4 exogenous expression for 7 days induced pluripotencyslightly as it was detected by significant enhancement in expression of nanog (p < 0.05).combinatorial administration of oct4 expressing vector and bix-01294, bay k8644 andrg-108 did not affect the expression of pluripotency and nsc markers, but vpa treatmentalong with oct4 exogenous expression induced nanog, klf4 and c-myc (p < 0.001). vpatreatment before the induction of exogenous oct4 was more effective and significantlyincreased the expression of endogenous oct4, nanog, klf4, c-myc (p < 0.01), pax6 andsox1 (p < 0.001).conclusion: these results suggest vpa as the best enhancer of pluripotency among thechemicals tested, especially when applied prior to pluripotency induction by oct4.