Insulin mutations impair beta-cell development in a patient-derived iPSC model of neonatal diabetes
- University of Helsinki, Finland
- University of Eastern Finland, Finland
Abstract
Insulin gene mutations are a leading cause of neonatal diabetes. They can lead to proinsulin misfolding and its retention in endoplasmic reticulum (ER). This results in increased ER-stress suggested to trigger beta-cell apoptosis. In humans, the mechanisms underlying beta-cell failure remain unclear. Here we show that misfolded proinsulin impairs developing beta-cell proliferation without increasing apoptosis. We generated induced pluripotent stem cells (iPSCs) from people carrying insulin (INS) mutations, engineered isogenic CRISPR-Cas9 mutation-corrected lines and differentiated them to beta-like cells. Single-cell RNA-sequencing analysis showed increased ER-stress and reduced proliferation in INS-mutant beta-like cells compared with corrected controls. Upon transplantation into mice, INS-mutant grafts presented reduced insulin secretion and aggravated ER-stress. Cell size, mTORC1 signaling, and respiratory chain subunits expression were all reduced in INS-mutant beta-like cells, yet apoptosis was not increased at any stage. Our results demonstrate that neonatal diabetes-associated INS-mutations lead to defective beta-cell mass expansion, contributing to diabetes development.
Data availability
Single cell RNA sequencing raw data was deposited in GEO under GSE115257Source data for single cell RNA sequencing as well as code scripts for analysis have been provided.
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The raw single-cell RNA sequencing data used in the studyNCBI Gene Expression Omnibus, GSE115257.
Article and author information
Author details
Funding
Suomen Akatemia
- Timo Otonkoski
Sigrid Juselius Foundation
- Timo Otonkoski
Novo Nordisk Foundation
- Timo Otonkoski
EU 7FP Integrated project BETACURE
- Timo Otonkoski
Diabetes Research Foundation
- Timo Otonkoski
Diabetes Wellness Foundation
- Diego Balboa
- Jonna Saarimäki-Vire
Biomedicum Helsinki Foundation
- Diego Balboa
INNODIA
- Timo Otonkoski
The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Ethics
Animal experimentation: Animal care and experiments were conducted as approved by the National Animal Experiment Board in Finland (ESAVI/9978/04.10.07/2014).
Human subjects: The Coordinating Ethics Committee of the Helsinki and Uusimaa Hospital District (no. 423/13/03/00/08) approved the patient informed consent for the derivation of the hiPSC lines used in this study: HEL71.4 and HEL107.2.
Reviewing Editor
- Anna L Gloyn, University of Oxford, United Kingdom
Publication history
- Received: May 20, 2018
- Accepted: November 6, 2018
- Accepted Manuscript published: November 9, 2018 (version 1)
- Version of Record published: December 14, 2018 (version 2)
Copyright
© 2018, Balboa et al.
This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.
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Insulin mutations impair beta-cell development in a patient-derived iPSC model of neonatal diabetes
eLife 7:e38519.
https://doi.org/10.7554/eLife.38519
Further reading
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- Stem Cells and Regenerative Medicine
Exploring how proliferation and maturation of beta-cells can be impaired after birth will shed light on the origins of various forms of diabetes.
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- Stem Cells and Regenerative Medicine
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