Using Human Induced Pluripotent Stem Cell Derived Organoids to Identify New Pathologies in Patients with PDX1 Mutations

Mansa Krishnamurthy, Division of Endocrinology; Cincinnati Children's Hospital Medical Center, University of Cincinnati College of Medicine, Cincinnati, OH; Center for Stem Cell & Organoid Medicine (CuSTOM); Cincinnati Children's Hospital Medical Center, University of Cincinnati College of Medicine, Cincinnati, OH.
Daniel O. Kechele, Division of Developmental Biology; Cincinnati Children's Hospital Medical Center, University of Cincinnati College of Medicine, Cincinnati, OH; Center for Stem Cell & Organoid Medicine (CuSTOM); Cincinnati Children's Hospital Medical Center, University of Cincinnati College of Medicine, Cincinnati, OH.
Taylor Broda, Division of Developmental Biology; Cincinnati Children's Hospital Medical Center, University of Cincinnati College of Medicine, Cincinnati, OH; Center for Stem Cell & Organoid Medicine (CuSTOM); Cincinnati Children's Hospital Medical Center, University of Cincinnati College of Medicine, Cincinnati, OH.
Xinghao Zhang, Division of Developmental Biology; Cincinnati Children's Hospital Medical Center, University of Cincinnati College of Medicine, Cincinnati, OH; Center for Stem Cell & Organoid Medicine (CuSTOM); Cincinnati Children's Hospital Medical Center, University of Cincinnati College of Medicine, Cincinnati, OH.
Jacob R. Enriquez, Division of Developmental Biology; Cincinnati Children's Hospital Medical Center, University of Cincinnati College of Medicine, Cincinnati, OH; Center for Stem Cell & Organoid Medicine (CuSTOM); Cincinnati Children's Hospital Medical Center, University of Cincinnati College of Medicine, Cincinnati, OH.
Heather A. McCauley, Division of Developmental Biology; Cincinnati Children's Hospital Medical Center, University of Cincinnati College of Medicine, Cincinnati, OH; Center for Stem Cell & Organoid Medicine (CuSTOM); Cincinnati Children's Hospital Medical Center, University of Cincinnati College of Medicine, Cincinnati, OH.
J Guillermo Sanchez, Division of Developmental Biology; Cincinnati Children's Hospital Medical Center, University of Cincinnati College of Medicine, Cincinnati, OH; Center for Stem Cell & Organoid Medicine (CuSTOM); Cincinnati Children's Hospital Medical Center, University of Cincinnati College of Medicine, Cincinnati, OH.
Kyle McCracken, Division of Developmental Biology; Cincinnati Children's Hospital Medical Center, University of Cincinnati College of Medicine, Cincinnati, OH; Center for Stem Cell & Organoid Medicine (CuSTOM); Cincinnati Children's Hospital Medical Center, University of Cincinnati College of Medicine, Cincinnati, OH.
Joseph Palermo, Division of Gastroenterology; Cincinnati Children's Hospital Medical Center, University of Cincinnati College of Medicine, Cincinnati, OH.
Anas Bernieh, Division of Pathology; Cincinnati Children's Hospital Medical Center, University of Cincinnati College of Medicine, Cincinnati, OH.
Margaret H. Collins, Division of Pathology; Cincinnati Children's Hospital Medical Center, University of Cincinnati College of Medicine, Cincinnati, OH.
Inas H. Thomas, Division of Pediatric Endocrinology; University of Michigan, Ann Arbor, MI.
Haley C. Neef, Division of Pediatric Gastroenterology; University of Michigan, Ann Arbor, MI.
Amer Heider, Division of Pathology; University of Michigan, Ann Arbor, MI.
Andrew Dauber, Division of Endocrinology, Children's National Hospital; Department of Pediatrics, The George Washington University School of Medicine and Health Sciences; Washington, DC.
James M. Wells, Division of Endocrinology; Cincinnati Children's Hospital Medical Center, University of Cincinnati College of Medicine, Cincinnati, OH; Division of Developmental Biology; Cincinnati Children's Hospital Medical Center, University of Cincinnati College of Medicine, Cincinnati, OH; Center for Stem Cell & Organoid Medicine (CuSTOM); Cincinnati Children's Hospital Medical Center, University of Cincinnati College of Medicine, Cincinnati, OH.

Document Type

Journal Article

Publication Date

7-5-2022

Journal

Gastroenterology

DOI

10.1053/j.gastro.2022.06.083

Keywords

CRISPR/Cas9; PDX1; metaplasia; organoids

Abstract

BACKGROUND AIMS: Two patients with homozygous mutations in PDX1 presented with pancreatic agenesis, chronic diarrhea and poor weight gain, the causes of which were not identified through routine clinical testing. We aim to perform a deep analysis of the stomach and intestine using organoids derived from induced pluripotent stem cells from PDX1 patients. METHODS: Gastric fundic, antral and duodenal organoids were generated using iPSC lines from a PDX1 patient and an isogenic iPSC line where the PDX1 point mutation was corrected. RESULTS: Patient-derived PDX1 antral organoids exhibited an intestinal phenotype, while intestinal organoids underwent gastric metaplasia with significant reduction in enteroendocrine cells. This prompted a re-examination of gastric and intestinal biopsies from both PDX1 patients, which recapitulated the organoid phenotypes. Moreover, antral biopsies also demonstrated increased parietal cells and lacked G-cells suggesting loss of antral identity. All organoid pathologies were reversed upon CRISPR-mediated correction of the mutation. CONCLUSION: These patients will now be monitored for the progression of metaplasia and gastrointestinal complications that might be related to the reduced gastric and intestinal endocrine cells. This study demonstrates the utility of organoids in diagnosing uncovered pathologies.

Department

Pediatrics

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