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Depolymerizing F-actin accelerates the exit from pluripotency to enhance stem cell-derived islet differentiation.

Nathaniel J Hogrebe¹, Mason D Schmidt¹, Punn Augsornworawat²

¹Division of Endocrinology, Metabolism and Lipid Research, Washington University School of Medicine, MSC 8127-057-08, 660 South Euclid Avenue, St. Louis, MO 63110, USA.

Biorxiv : the Preprint Server for Biology

|November 1, 2024

Summary

Cytoskeletal state influences human pluripotent stem cell differentiation. Disrupting actin polymerization with latrunculin A improves pancreatic islet development and beta cell function.

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Area of Science:

Stem cell biology
Developmental biology
Cellular signaling

Background:

Cytoskeletal dynamics play a role in cell fate decisions.
Understanding early differentiation cues is crucial for regenerative medicine.

Purpose of the Study:

To investigate the role of cytoskeletal state in human pluripotent stem cell (hPSC) differentiation.
To determine if manipulating the actin cytoskeleton can improve stem cell-derived islet (SC-islet) generation.

Main Methods:

hPSCs were subjected to directed differentiation protocols.
Latrunculin A (latA) was used to depolymerize F-actin during definitive endoderm formation.
Signaling pathway dynamics (Activin/Nodal, BMP, JNK-JUN, WNT) were analyzed.
SC-islets were generated and characterized for beta cell content, maturation, and function.

Main Results:

Depolymerizing F-actin with latA accelerated hPSC exit from pluripotency.
LatA treatment modulated key developmental signaling pathways.
This approach enhanced pancreatic progenitor identity and beta cell yield.
SC-islets showed improved insulin secretion and in vivo glucose control.
LatA treatment corrected differentiation failures in certain hPSC lines.

Conclusions:

Cytoskeletal state at the start of differentiation is critical for hPSC lineage specification.
Targeting actin dynamics offers a novel strategy to enhance SC-islet production.
This method improves beta cell function and corrects differentiation variability.