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

被引:228
作者
Hogrebe, Nathaniel J. [1 ]
Augsornworawat, Punn [1 ,2 ]
Maxwell, Kristina G. [1 ]
Velazco-Cruz, Leonardo [1 ]
Millman, Jeffrey R. [1 ,2 ]
机构
[1] Washington Univ, Sch Med, Div Endocrinol Metab & Lipid Res, St Louis, MO USA
[2] Washington Univ, Dept Biomed Engn, St Louis, MO 63110 USA
基金
美国国家卫生研究院;
关键词
IN-VITRO; EXPRESSION ANALYSIS; ACTIN DYNAMICS; HUMAN LIVER; PROGENITORS; LINEAGE; GENERATION; MATURATION; TRANSCRIPTION; DISEASE;
D O I
10.1038/s41587-020-0430-6
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Generation of pancreatic beta 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 beta (SC-beta) cells with improved in vitro and in vivo function. SC-beta 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. Generation of pancreatic beta cells from stem cells is enhanced by manipulating the cytoskeleton.
引用
收藏
页码:460 / +
页数:15
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