Open AccessTargeting the cytoskeleton to direct pancreatic differentiation of human pluripotent stem cells
Author(s) -
Nathaniel J. Hogrebe,
Punn Augsornworawat,
Kristina G. Maxwell,
Leonardo Velazco-Cruz,
Jeffrey R. Millman
Publication year - 2020
Publication title -
nature biotechnology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 15.358
H-Index - 445
eISSN - 1546-1696
pISSN - 1087-0156
DOI - 10.1038/s41587-020-0430-6
Subject(s) - induced pluripotent stem cell , microbiology and biotechnology , biology , stem cell , cytoskeleton , cellular differentiation , islet , actin cytoskeleton , transplantation , actin , enteroendocrine cell , cell , embryonic stem cell , insulin , medicine , endocrinology , endocrine system , biochemistry , gene , hormone
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.