Identification Of Long Non Coding Rnas Associated To Mesendoderm Differentiation And Cardiovascular Commitment Of Early Precursors Generated By Naive Mouse Embryonic Stem Cells

Introduction: Nitric oxide (NO) negatively regulates the zinc-finger E-box binding homeobox 1 and 2 (Zeb1, 2) factors via upregulation of miR-200 family members. Although Zeb1, 2 are well known inducers of epithelial-to-mesenchymal transition, an important step to heart formation, their role in mesendoderm differentiation is still poorly understood. In this study, we provide evidence that, in mouse embryonic stem cells (mES), the Zeb factors are downregulated by NO with important consequences on the expression of crucial mesendodermal genes and on a subset of NO-dependent long non-coding RNAs (lncRNAs). Methods and Results: mES, classically cultured in the presence of serum and LIF (L), synthesize NO only late during differentiation. Instead, mES kept in the presence of three inhibitors (3i), namely L, GSK3i and ERKi realize the so called “naive state”, express rapidly the endothelial nitric oxide synthase (eNOS) and generate detectable NO levels very early after release from stemness. In fact, as early a...