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HNF1A-MODY – Disease modeling in-a-dish

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 HNF1A-MODY) causes about 70% of all MODY cases. Diabetes is induced by lowering the amount of insulin. However, the clinical expression in HNF1A-MODY 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 HNF1A-MODY often develop diabetic microvascular complications, related to the endothelial dysfunction, 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 endothelial dysfunction in HNF1A-MODY could provide new approaches for patient management. In the current project we are using two of the most impactful techniques for disease modeling – induced pluripotent stem cells (iPSCs) and CRISPR/Cas9 gene editing
Our work is based on the usage of so-called isogenic lines that differ exclusively at the disease-causing mutation. Thus the individual variances could be diminished and subtle disease-relevant differences in monogenic diseases could be revealed. 

 

 

 

 

 

 

 

Using isogenic iPSCs lines we found that cells with heterozygous mutation in HNF1A gene have increased vascular permeability after stimulation with pro-inflammatory cytokine (Kachamakova-Trojanowska et al., 2019). And this effect was even more pronounced in the cells with biallelic mutation. Currently, we would like to dissect the molecular background of the observed increase of the vascular permeability and to reveal its possible relation to HNF1A mutation.

Research techniques:
Derivation of iPSCs from human somatic cells
Differentiation of iPSCs toward endothelial cells
CRISPR/Cas9 gene editing techniques
Western blot
Immunostaining 
Culture of cells under flow conditions (ibidi pump system)
Live imaging (Nanolive)
Gene expression
Flow cytometry analysis
Cell sorting techniques

 

Publications:
1. Skoczek et al., Maturity Onset Diabetes of the Young-New Approaches for Disease Modelling, Int J Mol Sci. 2021.
2. Kachamakova-Trojanowska et al., HIF-1 stabilization exerts anticancer effects in breast cancer cells in vitro and in vivo. Biochem Pharmacol. 2020 
3. Białopiotrowicz et al., Serine Biosynthesis Pathway Supports MYC-miR-494-EZH2 Feed-Forward Circuit Necessary to Maintain Metabolic and Epigenetic Reprogramming of Burkitt Lymphoma Cells. Cancers (Basel). 2020
4. Szade et al., Heme oxygenase-1 deficiency triggers exhaustion of hematopoietic stem cells. EMBO Rep. 2020
5. Rozga et al., Novel engineered TRAIL-based chimeric protein strongly inhibits tumor growth and bypasses TRAIL resistance. Int J Cancer. 2019
6. Kachamakova-Trojanowska et al., Human iPSCs-Derived Endothelial Cells with Mutation in HNF1A as a Model of Maturity-Onset Diabetes of the Young, Cells. 2019
7. Szade et al., Cobalt protoporphyrin IX increases endogenous G-CSF and mobilizes HSC and granulocytes to the blood. EMBO Mol Med. 2019
8. Nowak et al., Atorvastatin and Conditioned Media from Atorvastatin-Treated Human Hematopoietic Stem/Progenitor-Derived Cells Show Proangiogenic Activity In Vitro but Not In Vivo. Mediators Inflamm. 2019
9. Mucha et al., Development and characterization of a new inhibitor of heme oxygenase activity for cancer treatment. Arch Biochem Biophys. 2019
10. Šmíd et al., Heme Oxygenase-1 May Affect Cell Signalling via Modulation of Ganglioside Composition. Oxid Med Cell Longev. 2018.
11. Kachamakova-Trojanowska et al., Molecular profiling of regulatory T cells in pulmonary sarcoidosis. J Autoimmun. 2018
12. Mucha et al., Pharmacological versus genetic inhibition of heme oxygenase-1 - the comparison of metalloporphyrins, shRNA and CRISPR/Cas9 system. Acta Biochim Pol. 2018
13. Stepniewski et al., Heme oxygenase-1 affects generation and spontaneous cardiac differentiation of induced pluripotent stem cells. IUBMB Life. 2018

 

Funding:
2017-2021 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)” 

2021-2025 Project Sonata-Bis headed by Dr. Neli Kachamakova-Trojanowska: “Molecular background of microvascular complications in HNF1A-MODY patients: induced pluripotent stem cells as disease modeling tool

Open positions:
We are currently looking for a talented and enthusiastic postdoctoral researcher to join our group. Interested? 
Contact dr. Neli Kachamakova-Trojanowska (neli.kachamakova-trojanowska@uj.edu.pl).