Targeting the cytoskeleton to direct pancreatic differentiation of human pluripotent stem cells

Nathaniel J. Hogrebe, Punn Augsornworawat, Kristina G. Maxwell, Leonardo Velazco-Cruz, Jeffrey R. Millman

Research output: Contribution to journalArticlepeer-review

91 Scopus citations

Abstract

Generation of pancreatic β cells from human pluripotent stem cells (hPSCs) holds promise as a cell replacement therapy for diabetes. In this study, we establish a link between the state of the actin cytoskeleton and the expression of pancreatic transcription factors that drive pancreatic lineage specification. Bulk and single-cell RNA sequencing demonstrated that different degrees of actin polymerization biased cells toward various endodermal lineages and that conditions favoring a polymerized cytoskeleton strongly inhibited neurogenin 3-induced endocrine differentiation. Using latrunculin A to depolymerize the cytoskeleton during endocrine induction, we developed a two-dimensional differentiation protocol for generating human pluripotent stem-cell-derived β (SC-β) cells with improved in vitro and in vivo function. SC-β cells differentiated from four hPSC lines exhibited first- and second-phase dynamic glucose-stimulated insulin secretion. Transplantation of islet-sized aggregates of these cells rapidly reversed severe preexisting diabetes in mice at a rate close to that of human islets and maintained normoglycemia for at least 9 months.

Original languageEnglish
Pages (from-to)460-470
Number of pages11
JournalNature Biotechnology
Volume38
Issue number4
DOIs
StatePublished - Apr 1 2020

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