Studies of monogenic diabetes are particularly useful because we can gain insight into the molecular events of pancreatic β-cell failure. Maturity-onset diabetes of the young 1 (MODY1) is a form of monogenic diabetes caused by a mutation in the HNF4A gene. Human-induced pluripotent stem cells (hiPSCs) provide an excellent tool for disease modeling by subsequently directing differentiation toward desired pancreatic islet cells, but cellular phenotypes in terminally differentiated cells are notoriously difficult to detect. Re-creating a spatial (three-dimensional [3D]) environment may facilitate phenotype detection. We studied MODY1 by using hiPSC-derived pancreatic β-like patient and isogenic control cell lines in two different 3D contexts. Using size-adjusted cell aggregates and alginate capsules, we show that the 3D context is critical to facilitating the detection of mutation-specific phenotypes. In 3D cell aggregates, we identified irregular cell clusters and lower levels of structural proteins by proteome analysis, whereas in 3D alginate capsules, we identified altered levels of glycolytic proteins in the glucose sensing apparatus by proteome analysis. Our study provides novel knowledge on normal and abnormal function of HNF4A, paving the way for translational studies of new drug targets that can be used in precision diabetes medicine in MODY.
The two chosen 3D environments challenge the differentiating pancreatic β-like cells in unique ways, including the levels of oxygen delivery, nutrient supply, and cell-to-cell contact. The 3D cell aggregation/AggreWell context is preferred by several investigators in generating pancreatic β-like cells because it mimics the in vivo pancreatic islets in size, level of cell-to-cell contact, and the self-organized compact spheroid arrangement. In the 3D alginate encapsulation context, the cells are immobilized and lack cell-to-cell contact; however, the alginate gel-pore network still allows transport of oxygen and diffusion of nutrients and waste products. Furthermore, alginate encapsulation is suitable for future therapeutic transplantation and can also protect the cells in bioreactor culture systems.
In conclusion, our findings warrants a careful consideration and selection of an appropriate 3D context when studying stem cell models. Our study also provides a patient-specific diabetes model that can be further explored for potential new drug targets that can be used in precision diabetes medicine.
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Spatial Environment Affects HNF4A Mutation-Specific Proteome Signatures and Cellular Morphology in hiPSC-Derived β-Like Cells - PubMed (nih.gov)
Diabetes. 2022 Apr 1;71(4):862-869.
Manuel Carrasco, Chencheng Wang, Anne M Søviknes, Yngvild Bjørlykke, Shadab Abadpour, Joao A Paulo, Erling Tjora, Pål Njølstad, Jonas Ghabayen, Ingrid Nermoen, Valeriya Lyssenko, Simona Chera, Luiza M Ghila, Marc Vaudel, Hanne Scholz, Helge Ræder