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Cited by 373Pancreatic function and diabetesPluripotent Stem Cells ResearchDiabetes and associated disorders

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

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Hogrebe, Nathaniel J., Augsornworawat, Punn, Maxwell, Kristina G., Velazco-Cruz, Leonardo, Millman, Jeffrey R.·2020·Nature Biotechnology
DOI 10.1038/s41587-020-0430-6PMC PMC7274216
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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.

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  • A beta-like cell with behavior that fully recapitulates the physiology of a normal beta cell is yet to be realized.

    “While a beta-like cell with behavior that fully recapitulates the physiology of a normal beta cell is yet to be realized”

    From 100 years of insulin: celebrating the past, present and future of diabetes therapy (2021) · cites this paper as [136]

    The Results (A planar SC-β cell differentiation method; page not available) show that the authors’ cells differed from human islets: MAFA and UCN3 expression remained low, and transplanted cells restored glucose levels slightly more slowly than human islet transplants. This supports the narrower point that their cells did not fully match normal β cells, but the paper does not establish that no β-like cell has yet done so. (LLM, conf=0.82)

    Experimental — accuracy not yet measured

  • There has been a steady series of improvements to directed differentiation strategies for generating beta-like cells over the past 20 years.

    “there has been a steady series of improvements to directed differentiation strategies over the past 20 years135–138.”

    From 100 years of insulin: celebrating the past, present and future of diabetes therapy (2021) · cites this paper as [136]

    The Introduction describes earlier stepwise strategies for differentiating stem cells toward pancreatic β cells, and the Results report improved performance with this paper’s planar protocol. These passages support improvements to differentiation strategies, but they do not establish a steady series of improvements over the claimed 20-year period. (LLM, conf=0.85)

    Experimental — accuracy not yet measured

  • With less optimized protocols or cell lines, there is potential to retain progenitor cells in the final islet clusters as well as produce other endodermal cell types, such as those of hepatic origin.

    “With less optimized protocols or cell lines, there is also the potential to retain progenitor cells in the final islet clusters as well as produce other endodermal cell types, such as those of hepatic origin.39,58”

    From Developments in stem cell-derived islet replacement therapy for treating type 1 diabetes (2023) · cites this paper as [39]

    The Introduction (page unavailable) says endodermal progenitors can produce non-pancreatic lineages, including hepatocytes. The Results section on cytoskeletal fate selection (page unavailable) also finds pancreatic progenitors and undefined endoderm under some differentiation conditions. These passages support the possibility of alternative cell fates, but they do not establish that less-optimized protocols or cell lines retain progenitors or produce hepatic cells in final islet clusters. (LLM, conf=0.84)

    Experimental — accuracy not yet measured

  • These SC-β cells have been shown to improve glycemic control in diabetic mice.

    “These cells have been shown to improve glycemic control in diabetic mice, highlighting their potential utility in vivo.36,37,39–44”

    From Developments in stem cell-derived islet replacement therapy for treating type 1 diabetes (2023) · cites this paper as [39]

  • Significant progress has been made at improving the function of SC-islets when compared to the original protocols.

    “To this end, significant progress has been made at improving the function of SC-islets when compared to the original protocols,39,40,44,48,49”

    From Developments in stem cell-derived islet replacement therapy for treating type 1 diabetes (2023) · cites this paper as [39]

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Research works printed but not linked: 51

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Cited by

Sorted by citations

Authors
Hogrebe, N. J., Augsornworawat, P., Maxwell, K. G., Velazco-Cruz, L., & Millman, J. R.
Affiliations
Washington University in St. Louis (US)
Corresponding author
Millman, Jeffrey R.
ORCID
Venue
Nature Biotechnology
Publisher
Springer Science and Business Media LLC
Published
2020-02-24
Volume
38
Issue
4
Pages
460-470
ISSN
  • 0733-222X (ISSN-L)
  • 1087-0156
  • 1546-1696
DOI
10.1038/s41587-020-0430-6
PMC
PMC7274216
OpenAlex
W3007716699
Cited by
373
References
13
Citations by year
2026: 42 · 2025: 58 · 2024: 68 · 2023: 57 · 2022: 60 · 2021: 56 · 2020: 32
FWCI
46.84
Type
journal article
Language
en
Open access
Licence
Locations
  • Nature Biotechnology (publishedVersion)
  • PubMed (publishedVersion)
  • PubMed Central (acceptedVersion)
Funders
  • U.S. Department of Health & Human Services | NIH | Center for Information Technology (T32DK007120, T32DK108742, R25GM103757)
  • JDRF (5-CDA-2017-391-A-N)
  • U.S. Department of Health & Human Services | NIH | Office of Extramural Research, National Institutes of Health (5R01DK114233)
Topics
  • Health Sciences › Medicine › Surgery › Pancreatic function and diabetes
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Pluripotent Stem Cells Research
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Diabetes and associated disorders
Keywords
pancreatic beta cells, human pluripotent stem cells, actin cytoskeleton, actin polymerization, latrunculin A, endocrine differentiation, neurogenin 3, pancreatic transcription factors, glucose-stimulated insulin secretion, islet transplantation, single-cell RNA sequencing
MeSH terms
Actins, Animals, Cell Differentiation, Cells, Cultured, Cytoskeleton, Diabetes Mellitus, Endoderm, Humans, Nerve Tissue Proteins, Trans-Activators, Homeodomain Proteins, Cell Lineage, Bridged Bicyclo Compounds, Heterocyclic, Pluripotent Stem Cells, Tubulin Modulators, Insulin-Secreting Cells, Mice, Basic Helix-Loop-Helix Transcription Factors, Basic Helix-Loop-Helix Proteins, Thiazolidines, Cell Engineering, Cell- and Tissue-Based Therapy

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