Pancreatic Differentiation of Stem Cells Reveals Pathogenesis of a Syndrome of Ketosis-Prone Diabetes

Authors

Diane Yang
Sanjeet Patel
Wojciech J Szlachcic
Jolanta Chmielowiec
Diane Scaduto
Nagireddy Putluri
Arun Sreekumar
James Suliburk
Michael Metzker
Ashok Balasubramanyam
Malgorzata Borowiak

Publication Date

10-1-2021

Journal

Diabetes

DOI

10.2337/db20-1293

PMID

34344789

PMCID

PMC8576504

PubMedCentral® Posted Date

8-3-2021

PubMedCentral® Full Text Version

Post-print

Published Open-Access

yes

Keywords

Adult, Cell Differentiation, Cells, Cultured, DNA Mutational Analysis, Diabetes Mellitus, Type 1, Gene Expression Regulation, Homeodomain Proteins, Humans, Induced Pluripotent Stem Cells, Insulin-Secreting Cells, Male, Pancreas, Syndrome, Trans-Activators

Abstract

Genetic analysis of an adult patient with an unusual course of ketosis-prone diabetes (KPD) and lacking islet autoantibodies demonstrated a nucleotide variant in the 5′-untranslated region (UTR) of PDX1, a β-cell development gene. When differentiated to the pancreatic lineage, his induced pluripotent stem cells stalled at the definitive endoderm (DE) stage. Metabolomics analysis of the cells revealed that this was associated with leucine hypersensitivity during transition from the DE to the pancreatic progenitor (PP) stage, and RNA sequencing showed that defects in leucine-sensitive mTOR pathways contribute to the differentiation deficiency. CRISPR/Cas9 manipulation of the PDX1 variant demonstrated that it is necessary and sufficient to confer leucine sensitivity and the differentiation block, likely due to disruption of binding of the transcriptional regulator NFY to the PDX1 5′-UTR, leading to decreased PDX1 expression at the early PP stage. Thus, the combination of an underlying defect in leucine catabolism characteristic of KPD with a functionally relevant heterozygous variant in a critical β-cell gene that confers increased leucine sensitivity and inhibits endocrine cell differentiation resulted in the phenotype of late-onset β-cell failure in this patient. We define the molecular pathogenesis of a diabetes syndrome and demonstrate the power of multiomics analysis of patient-specific stem cells for clinical discovery.

Comments

This article has been corrected. See Diabetes. 2023 Apr 17;72(6):821.