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Related Experiment Video

Updated: Aug 22, 2025

Mixed Primary Cultures of Murine Small Intestine Intended for the Study of Gut Hormone Secretion and Live Cell Imaging of Enteroendocrine Cells
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Mapping prohormone processing by proteases in human enteroendocrine cells using genetically engineered organoid

Joep Beumer1, Julia Bauzá-Martinez2, Tim S Veth2

  • 1Hubrecht Institute, Royal Netherlands Academy of Arts and Sciences (KNAW) and University Medical Center (UMC) Utrecht, Oncode Institute, 3584 CT Utrecht, The Netherlands.

Proceedings of the National Academy of Sciences of the United States of America
|November 7, 2022
PubMed
Summary

This study uses human gut organoids to investigate how enteroendocrine cell (EEC) hormones are processed. Researchers identified key proteases involved in generating bioactive peptides, advancing our understanding of hormone production and related diseases.

Keywords:
CRISPR-Cas9enteroendocrine cellsintestinal organoidspeptidomicsprohormone processing

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Area of Science:

  • Gastroenterology and Endocrinology
  • Molecular Biology
  • Proteomics

Background:

  • Enteroendocrine cells (EECs) produce hormones regulating appetite and insulin release.
  • Hormone production involves proteolytic processing of proproteins into bioactive peptides.
  • Mutations in EEC proteases are linked to endocrinopathies, but studying these cells is difficult due to their rarity and lack of in vitro models.

Purpose of the Study:

  • To develop and utilize an in vitro model for studying intestinal prohormone processing in human EECs.
  • To comprehensively map peptide hormones produced by human EECs and the roles of specific proteases in their generation.

Main Methods:

  • Human gut organoids were engineered using CRISPR-Cas9 to modify EEC-expressed endopeptidase and exopeptidase genes.
  • Mass spectrometry-based analyses were employed to monitor peptide processing.
  • Glucagon production in EECs was identified and stimulated via bone morphogenic protein (BMP) signaling.

Main Results:

  • The study successfully mapped substrates and products of major EEC endo- and exopeptidases.
  • Specific proteases' roles in generating bioactive peptide hormones were defined.
  • Glucagon production in intestinal EECs was confirmed and linked to BMP signaling.

Conclusions:

  • Human gut organoids provide a viable model for studying EEC prohormone processing.
  • This research offers a comprehensive description of peptide hormones produced by human EECs.
  • The study elucidates the critical functions of specific proteases in the generation of these vital hormones.