# Effects of Neurogenin 3 Induction on Endocrine Differentiation and Delamination in Adult Human Pancreatic Ductal Organoids

## TL;DR

This study explores how inducing Neurogenin 3 in adult human pancreatic organoids can lead to the formation of endocrine cells, offering insights into potential diabetes treatments.

## Contribution

The study demonstrates that NEUROG3 induction in adult pancreatic organoids can trigger endocrine differentiation and delamination, similar to embryonic development.

## Key findings

- Doxycycline-induced NEUROG3 expression leads to CHGA+ endocrine progenitor cells in organoids.
- Adding thyroid hormone T3 and AXL inhibitor R428 improves differentiation efficiency.
- Modifying Notch, YAP, and EGFR signaling increases NEUROG3 expression and endocrine cell formation.

## Abstract

Diabetes mellitus is characterized by the loss of pancreatic insulin-secreting β-cells in the Islets of Langerhans. Understanding the regenerative potential of human islet cells is relevant in the context of putative restoration of islet function after damage and novel islet cell replacement therapies. Adult human pancreatic tissue can be cultured as three-dimensional organoids with the capacity for long-term expansion and the promise of endocrine cell formation. Here, we characterize the endocrine differentiation potential of human adult pancreatic organoids. Because exocrine-to-endocrine differentiation is dependent on the expression of Neurogenin 3 (NEUROG3), we first generated NEUROG3-inducible organoid lines. We show that doxycycline-induced NEUROG3 expression in the organoids leads to the formation of chromogranin A positive (CHGA+) endocrine progenitor cells. The efficiency of this differentiation was improved with the addition of thyroid hormone T3 and the AXL inhibitor R428. Further, compound screening demonstrated that modifying the pivotal embryonic endocrine pancreas signalling pathways driven by Notch, YAP, and EGFR led to increased NEUROG3 expression in organoids. In a similar fashion to embryonic development, adult ductal cells delaminated from the organoids after NEUROG3 induction. Thus, mechanisms in islet (re)generation including the initiation of endocrine differentiation and delamination can be achieved by NEUROG3 induction.