A method for the generation of human stem cell-derived alpha cells

Quinn P Peterson,Maria Sörhede-Winzell,Björn Tyrberg,Adrian Veres,Douglas A Melton,Aleksey V Matveyenko,Quan Zhou,Michael Q Slama,Patrik Rorsman,Jennifer H R Kenty,Shaimaa Hassoun,Lihua Chen,Haiqiang Dou,Caden D Duffy,Matthew R Brown

doi:10.1038/s41467-020-16049-3

Abstract

The generation of pancreatic cell types from renewable cell sources holds promise for cell replacement therapies for diabetes. Although most effort has focused on generating pancreatic beta cells, considerable evidence indicates that glucagon secreting alpha cells are critically involved in disease progression and proper glucose control. Here we report on the generation of stem cell-derived human pancreatic alpha (SC-alpha) cells from pluripotent stem cells via a transient pre-alpha cell intermediate. These pre-alpha cells exhibit a transcriptional profile similar to mature alpha cells and although they produce proinsulin protein, they do not secrete significant amounts of processed insulin. Compound screening identified a protein kinase c activator that promotes maturation of pre-alpha cells into SC-alpha cells. The resulting SC-alpha cells do not express insulin, share an ultrastructure similar to cadaveric alpha cells, express and secrete glucagon in response to glucose and some glucagon secretagogues, and elevate blood glucose upon transplantation in mice.

Highlights

The generation of pancreatic cell types from renewable cell sources holds promise for cell replacement therapies for diabetes
Diabetes has been a leading candidate for this approach because a single-cell type, the beta cell, plays such a critical role in the disease. This led to a focus on the generation of pancreatic beta cells and much progress has been made, there are potential advantages in adding other cell types to treat diabetes
Paracrine signaling from alpha cells establishes a set point for insulin secretion[48], and glucagon is important for beta cell regeneration in a zebrafish model[49]

Summary

Introduction

The generation of pancreatic cell types from renewable cell sources holds promise for cell replacement therapies for diabetes. We report on the generation of stem cell-derived human pancreatic alpha (SC-alpha) cells from pluripotent stem cells via a transient pre-alpha cell intermediate These pre-alpha cells exhibit a transcriptional profile similar to mature alpha cells and they produce proinsulin protein, they do not secrete significant amounts of processed insulin. In 2011, Rezania et al.[15] reported a protocol for generating glucagon-positive cells that exhibited some glucagon secretion in vitro; upon transplantation of 1.9 million cells into mice, these cells had limited physiological effects. Despite these early efforts to generate glucagon-positive cells, the production of alpha cells has not been reproduced nor widely adopted by the field

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Conclusion