Inhibition of DYRK1A disrupts neural lineage specificationin human pluripotent stem cells.

Stephanie F Bellmaine,Spencer J Williams,Dmitry A Ovchinnikov,Andrew G Elefanty,Martin Pera,David T Manallack,Claire E Cuddy,Edouard G Stanley,Ernst J Wolvetang

doi:10.7554/elife.24502

Abstract

Genetic analysis has revealed that the dual specificity protein kinase DYRK1A has multiple roles in the development of the central nervous system. Increased DYRK1A gene dosage, such as occurs in Down syndrome, is known to affect neural progenitor cell differentiation, while haploinsufficiency of DYRK1A is associated with severe microcephaly. Using a set of known and newly synthesized DYRK1A inhibitors, along with CRISPR-mediated gene activation and shRNA knockdown of DYRK1A, we show here that chemical inhibition or genetic knockdown of DYRK1A interferes with neural specification of human pluripotent stem cells, a process equating to the earliest stage of human brain development. Specifically, DYRK1A inhibition insulates the self-renewing subpopulation of human pluripotent stem cells from powerful signals that drive neural induction. Our results suggest a novel mechanism for the disruptive effects of the absence or haploinsufficiency of DYRK1A on early mammalian development, and reveal a requirement for DYRK1A in the acquisition of competence for differentiation in human pluripotent stem cells.

Highlights

The dual-specificity tyrosine-phosphorylation-regulated (Dyrk) kinases belong to a family collectively referred to as CMGC kinases (Aranda et al, 2011) that includes cyclin dependent kinases, mitogen associated protein kinases, glycogen synthase kinases, and cyclin dependent-like kinases The distinguishing biochemical features of Dyrk kinases are their ability to phosphorylate serine, threonine and tyrosine residues, and the autoregulation of their kinase activity through tyrosine phosphorylation
We further showed that stable knockdown of DYRK1A and DYRK2 caused a modest increase in the plating efficiency of human embryonic stem cells (hESC), but we did not establish whether this effect was related to enhancement of attachment and survival, or to inhibition of differentiation
We present evidence that the indole compound ID-8 and a series of related molecules act to inhibit the neural specification of hESC through inhibition of DYRK1A

Summary

Introduction

The dual-specificity tyrosine-phosphorylation-regulated (Dyrk) kinases belong to a family collectively referred to as CMGC kinases (Aranda et al, 2011) that includes cyclin dependent kinases, mitogen associated protein kinases, glycogen synthase kinases, and cyclin dependent-like kinases The distinguishing biochemical features of Dyrk kinases are their ability to phosphorylate serine, threonine and tyrosine residues, and the autoregulation of their kinase activity through tyrosine phosphorylation. The Class 1 kinase DYRK1A has been implicated in a diverse variety of biological processes, including central nervous system. Developmental Biology and Stem Cells development, Down syndrome, cancer, beta cell proliferation and diabetes, and Alzheimer’s disease, and the discovery of novel DYRK1A inhibitors has been a goal of many recent studies (Abbassi et al, 2015; Aranda et al, 2011; Shen et al, 2015; Smith et al, 2012; Stotani et al, 2016). DYRK1A has multiple roles in central nervous system development (Tejedor and Hammerle, 2011). DYRK1A gene dosage affects later stages of neurogenesis, including neuronal dendritogenesis (Benavides-Piccione et al, 2005; Gockler et al, 2009). DYRK1A has been implicated in tau protein phosphorylation in the pathogenesis of Alzheimer’s disease (Coutadeur et al, 2015)

Methods

Results

Conclusion