Targeting the cytoskeleton to direct pancreatic differentiation of human pluripotent stem cells.
Nathaniel J HogrebePunn AugsornworawatKristina G MaxwellLeonardo Velazco-CruzJeffrey R MillmanPublished in: Nature biotechnology (2020)
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.
Keyphrases
- pluripotent stem cells
- induced apoptosis
- single cell
- endothelial cells
- cell cycle arrest
- induced pluripotent stem cells
- type diabetes
- cardiovascular disease
- cell death
- signaling pathway
- rna seq
- transcription factor
- poor prognosis
- adipose tissue
- drug delivery
- cell proliferation
- deep learning
- pi k akt
- early onset
- artificial intelligence
- machine learning
- weight loss
- drug induced
- skeletal muscle
- cell therapy
- big data
- insulin resistance
- diabetic rats