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A High-content In Vitro Pancreatic Islet β-cell Replication Discovery Platform

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Published on: July 16, 2016

A Pdx-1-Regulated Soluble Factor Activates Rat and Human Islet Cell Proliferation.

Heather L Hayes^1,2, Lu Zhang^1,2, Thomas C Becker^1,3

¹Sarah W. Stedman Nutrition and Metabolism Center and Duke Molecular Physiology Institute, Duke University Medical Center, Durham, North Carolina, USA.

Molecular and Cellular Biology

|September 14, 2016

Summary

The homeodomain transcription factor Pdx-1 promotes islet cell proliferation through secreted factors. Pdx-1 induces inhibin beta-B, which stimulates both rat and human islet cell growth.

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Area of Science:

Endocrinology
Molecular Biology
Developmental Biology

Background:

Pdx-1 is crucial for pancreas and islet development, β-cell function, and survival.
Previous studies indicated Pdx-1 overexpression stimulates islet cell proliferation, but the underlying mechanism was unknown.

Purpose of the Study:

To elucidate the mechanism by which Pdx-1 overexpression stimulates islet cell proliferation.
To identify the specific factors and pathways involved in Pdx-1-mediated islet cell growth.

Main Methods:

Overexpression of Pdx-1 in rat and human islets using cell-specific promoters (e.g., rat insulin promoter [RIP]).
Transwell co-culture experiments to assess non-cell-autonomous effects.
Microarray and gene ontology (GO) analysis to identify Pdx-1-responsive genes.
Overexpression of identified genes (e.g., inhibin beta-B [Inhbb]) and addition of recombinant proteins (e.g., activin B).
Assessment of activin receptor involvement (RIIA and RIIB).

Main Results:

Pdx-1 overexpression triggers islet cell proliferation via a non-cell-autonomous mechanism mediated by soluble factors.
Inhibin beta-B (Inhbb) was identified as a Pdx-1-responsive gene.
Overexpression of Inhbb or addition of activin B stimulated proliferation of rat and human islet cells, including β cells.
Activin receptors RIIA and RIIB were necessary for the full proliferative effects of Pdx-1 in rat islets.
In human islets, Inhbb overexpression enhanced Pdx-1-stimulated proliferation.

Conclusions:

Pdx-1 induces the secretion of soluble factors, such as Inhbb, that are sufficient to stimulate islet cell proliferation.
This study reveals a novel mechanism involving Pdx-1, Inhbb, and activin signaling in regulating islet cell growth in both rat and human models.
The findings highlight a non-cell-autonomous role for Pdx-1 in promoting islet regeneration or expansion.