FOXO1 inhibition yields functional insulin-producing cells in human gut organoid cultures

Abstract

Generation of surrogate sources of insulin-producing β-cells remains a goal of diabetes therapy. While most efforts have been directed at differentiating embryonic or induced pluripotent stem (iPS) cells into β-like-cells through endodermal progenitors, we have shown that gut endocrine progenitor cells of mice can be differentiated into glucose-responsive, insulin-producing cells by ablation of transcription factor Foxo1. Here we show that FOXO1 is present in human gut endocrine progenitor and serotonin-producing cells. Using gut organoids derived from human iPS cells, we show that FOXO1 inhibition using a dominant-negative mutant or lentivirus-encoded shRNA promotes generation of insulin-positive cells that express all markers of mature pancreatic β-cells, release C-peptide in response to secretagogues, and survive in vivo following transplantation into mice. The findings raise the possibility of using gut-targeted FOXO1 inhibition or gut organoids as a source of insulin-producing cells to treat human diabetes.

Advances in β cell replacement and regeneration strategies for treating diabetes

“…Similarly, ablation of the transcription factor FOXO1 in endocrine progenitors of the intestine results in the generation of insulin-expressing cells that are able to reverse hyperglycemia in mice (36). This finding has been reproduced in human gut organoids (23), suggesting that FOXO1 inhibition in gut organoids could offer a source of insulin-producing cells to treat human diabetes. As human gastrointestinal tissue is readily accessible by noninvasive techniques, ex vivo reprogramming of gastrointestinal cells could be a source of autologous β cells for implantation in patients with diabetes.…”

Reprogrammed Stomach Tissue as a Renewable Source of Functional β Cells for Blood Glucose Regulation

“…FoxO1 deletion also leads to formation of insulin + cells in the intestine, suggesting a therapeutic path toward inducing insulin + cells in situ (Bouchi et al, 2014; Talchai et al, 2012a). Our approach offers several advantages.…”

“…Development of gut enteroendocrine and pancreatic endocrine cells also depends on common critical factors, such as Ngn3 (also known as Neurog3 ) (Gu et al, 2002; Jenny et al, 2002; Lee et al, 2002). Recent studies showed that intestinal cells could be converted into insulin + cells with either endocrine-specific deletion of FoxO1 or ubiquitous expression of NPM reprogramming factors ( Ngn3 , Pdx1 , and Mafa ) (Bouchi et al, 2014; Chen et al, 2014; Talchai et al, 2012a). Although these studies revealed the feasibility of deriving β-like cells from the intestine, critical barriers remain in developing these approaches into future regenerative therapies.…”

Human Enteroids/Colonoids and Intestinal Organoids Functionally Recapitulate Normal Intestinal Physiology and Pathophysiology

“…Modifications to established protocols are likely to uncover new conditions required for specific aspects of division and differentiation of these preparations and reveal new translational applications. A recent example is the demonstration that inhibition of FOXO1 in human gut organoid cultures yields functional insulin-producing cells (60). Detailed documentation and comparisons will be necessary to properly define the genetic, biochemical, and physiologic aspects of the resulting enteroids and organoids.…”