Facilitation of Human Induced Pluripotent Stem (iPS) Cell Differentiation to Endoderm with a Novel Histone Deacetylase (HDAC) Inhibitor

Abstract

The use of patient iPS-derived cells for lung regeneration has emerged as a potential tissue engineering strategy for patients with end-stage lung diseases. The ability to efficiently derive lung cells from iPS cells would greatly facilitate the engineering process. Histone deacetylase inhibitors (HDACi) alter gene transcription by inhibiting the deacetylation of lysine residues and have been reported to induce differentiation of cancer stem cells (Botrugno, Santoro, & Minucci, 2009). We are evaluating the ability of a novel HDACi that is an analogue of suberoylanilide hydroxamic acid (SAHA), called SMAHA (Chen, Wilson, Jayaram, & Pankiewicz, 2007), to facilitate differentiation of iPS cells into definitive endoderm. Human iPS cells were generated as previously described (Hirai, Katoku-Kikyo, Karian, Firpo, & Kikyo, 2012). Definitive endoderm differentiation was induced using an established protocol (Mou et al., 2012a) with and without SMAHA at concentrations ranging between 0.0025 and 0.1 µM for 4 days of culture. Gene expression of the pluripotency markers NANOG and OCT-4, and endoderm markers FOXA2 and SOX 17 were assessed by RT-qPCR. The efficacy of SMAHA for inducing iPSCs into definitive endoderm is still under investigation. Preliminary results demonstrated some promise of endoderm induction for iPSCs exposed to SMAHA versus those that were not.