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HNF1A-MODY - Disease-in-a-dish

Diabetes is a global health issue affecting children, adolescents, and adults. According to the World Health Organization, approximately 180 million people worldwide currently have type 2 diabetes and this number is estimated to double by 2030 year. With up to 75% of mortality in diabetic patients arising from vascular diseases, this is an issue of utmost social importance. Many studies have established that endothelial dysfunction (ED) is not only initiator but could also be an important factor in the progression of vascular diseases. ED could develop as diabetes consequence or as recently shown could precede the development of diabetes.

Monogenic forms of diabetes are invaluable "human models" that have contributed to our understanding of the pathophysiological basis of common diabetes type 1 and type 2 (T1D and T2D). Maturity onset diabetes of the young (MODY) is caused by a mutation in a single gene and leads to diabetes under the age of 25. The mutations are inheritable, and any child have 50% chance of getting the same mutation as a parent. The mutations in HNF1A gene (leading to MODY3) causes about 70% of MODY cases. Diabetes is induced by lowering the amount of insulin. However, the clinical expression in MODY3 patients is quite variable even within the same family. Some can develop hyperglycemia, whereas others can be normoglycemic at the same age. It was shown that patients with MODY3 have diabetic microvascular complications, related to ED, however it is not clear whether these complications are result of the hyperglycemia or due to the genetic mutation in HNF1A gene.

Therefore, an improved understanding of the mechanisms and causes of ED could provide new approaches for MODY3 patient management. In the current project we intend to use two of the most impactful techniques for disease modeling – generation of induced pluripotent stem cells (iPSCs) and CRISPR/Cas9 gene editing method. The generation of isogenic pairs of disease-specific and control iPSC lines that differ exclusively at the disease-causing mutation would be used to control the individual variances and define the subtle disease-relevant differences in monogenic diseases.

It is worth mentioning that the combination of iPSCs and genome editing provides an unprecedented opportunity to study the fundamental principles of cell biology. iPSCs provide well-defined source of tissue-specific cells and are invaluable disease modeling tools. The derived iPSC lines, together with their respective isogenic controls will be further differentiated toward endothelial cells (ECs) and the functionality of these cells would be checked using multiple techniques. The expression of different endothelial markers on gene and protein level together with the production of important for the endothelial function compounds like nitric oxide, endothelin-1, reactive oxygen species, cytokines could be evaluated. Moreover, the iPSC-derived ECs could be cultured in shear stress conditions in order to recapitulate the in vivo blood flow stimulation. Under normal physiological conditions the endothelium is in a quiescent and anti-inflammatory state, however in the presence of risk factors it can be activated to express adhesion molecules like vascular cell adhesion molecule-1 and intracellular adhesion molecule-1, so the “activated” status of the cells could be evaluated in normal and pro-inflammatory conditions.


2018-2021 Project OPUS headed by Dr. Neli Kachamakova-Trojanowska:“Editing of HNF1A gene in human induced pluripotent stem cells with CRISPR/Cas9 for disease modeling of endothelial (dys)function in maturity onset diabetes of the young (MODY)”