NAMPT/SIRT2-mediated inhibition of the p53-p21 signaling pathway is indispensable for maintenance and hematopoietic differentiation of human iPS cells

Yun Xu,Masoud Nasri,Benjamin Dannenmann,Perihan Mir,Azadeh Zahabi,Karl Welte,Tatsuya Morishima,Julia Skokowa

doi:10.1186/s13287-021-02144-9

Yun Xu, Masoud Nasri + Show 6 more

Open Access

https://doi.org/10.1186/s13287-021-02144-9

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Abstract

BackgroundNicotinamide phosphoribosyltransferase (NAMPT) regulates cellular functions through the protein deacetylation activity of nicotinamide adenine dinucleotide (NAD+)-dependent sirtuins (SIRTs). SIRTs regulate functions of histones and none-histone proteins. The role of NAMPT/SIRT pathway in the regulation of maintenance and differentiation of human-induced pluripotent stem (iPS) cells is not fully elucidated.MethodsWe evaluated the effects of specific inhibitors of NAMPT or SIRT2 on the pluripotency, proliferation, survival, and hematopoietic differentiation of human iPS cells. We also studied the molecular mechanism downstream of NAMPT/SIRTs in iPS cells.ResultsWe demonstrated that NAMPT is indispensable for the maintenance, survival, and hematopoietic differentiation of iPS cells. We found that inhibition of NAMPT or SIRT2 in iPS cells induces p53 protein by promoting its lysine acetylation. This leads to activation of the p53 target, p21, with subsequent cell cycle arrest and induction of apoptosis in iPS cells. NAMPT and SIRT2 inhibition also affect hematopoietic differentiation of iPS cells in an embryoid body (EB)-based cell culture system.ConclusionsOur data demonstrate the essential role of the NAMPT/SIRT2/p53/p21 signaling axis in the maintenance and hematopoietic differentiation of iPS cells.

Highlights

Understanding the mechanisms underlying the maintenance and hematopoietic development of induced pluripotent stem cells is essential for the establishment of efficient protocols for iPS cell generation, ex vivo blood cell formation, and the identification of new treatment options for benign and oncogenic hematological disorders. protocols for iPS cell generation are well established, there is still room for improvement in terms of the efficient generation of high-quality iPS cells
Nicotinamide phosphoribosyltransferase (NAMPT) and SIRT2 are important for the embryoid body (EB)-based hematopoietic differentiation of iPS cells
We found that treatment with FK866 or AC93253 caused a concentration-dependent decrease in the absolute number of iPS cells (Fig. 1c)

Summary

Introduction

Understanding the mechanisms underlying the maintenance and hematopoietic development of induced pluripotent stem (iPS) cells is essential for the establishment of efficient protocols for iPS cell generation, ex vivo blood cell formation, and the identification of new treatment options for benign and oncogenic hematological disorders. protocols for iPS cell generation are well established, there is still room for improvement in terms of the efficient generation of high-quality iPS cells. Protein deacetylation triggered by NAMPT (nicotinamide phosphoribosyltransferase) and downstream NAD+-dependent sirtuins (SIRT) is important for myeloid differentiation and leukemogenic transformation of hematopoietic cells through regulation of the CCAAT/enhancer-binding proteins C/EBPα and C/EBPß, the serine/threonine kinase AKT, the tumor-suppressor p53, and the forkhead box transcription factor FOXO3 [8,9,10,11,12]. In these studies, SIRT1 and SIRT2, members of the SIRT family of NAD+dependent class III histone deacetylases [13], were found to activate target proteins in hematopoietic cells upon NAMP T activation. The role of NAMPT/SIRT pathway in the regulation of maintenance and differentiation of human-induced pluripotent stem (iPS) cells is not fully elucidated

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