Background
Patients with antithrombin deficiency are at increased risk of thrombosis, and many develop thrombotic events even while receiving anticoagulation therapy. Antithrombin deficiency is caused by mutations in the gene encoding antithrombin, SERPINE1. Our group has developed a method to generate liver organoids from induced pluripotent stem cells (iPSCs) that recapitulate key physiological functions of the liver, including antithrombin production.
Aims of the project
The overall aim of the project is to determine whether correction of disease-causing mutations in SERPINE1 can restore antithrombin production in liver cells. Specifically, we will:
1. Correct a severe SERPINE1 mutation in iPSCs from a patient using CRISPR/Cas9 and evaluate the impact on antithrombin expression and function.
2. Differentiate patient-derived, gene-corrected iPSCs into liver organoids and assess whether antithrombin production is normalized.
Methods
In this project, blood samples have already been collected from patients with antithrombin deficiency and reprogrammed into iPSCs. The next steps involve gene correction, differentiating these iPSCs into liver organoids and characterizing antithrombin production and function.
The project offers hands-on training in:
- Stem cell culture and hepatic organoid differentiation
- Molecular biology techniques, including RNA isolation, qPCR, Western blotting, ELISA
- CRISPR/Cas9-based gene editing and validation of gene correction
About the group
The research group is part of the Department of Hematology at Oslo University Hospital (OUS) and shares laboratory and office facilities with five other research groups. The environment is inclusive and positive, with researchers ranging from master’s students to senior scientists. The group has extensive experience with iPSC technology, organoid models, and CRISPR/Cas9 gene editing, and has previously conducted similar studies using cells from patients with bleeding disorders. All members are highly collaborative and supportive, providing a strong framework for learning and professional development.
Supervisors
Maria Eugenia Chollet, Giacomo Roman, Benedicte Stavik. Internal supervisor Rafal Ciosk
References
1) Roman G, Lauritzen KH, Harrison SP, Bhattacharya A, Andresen MS, Mowinckel MC, Smolková B, Henriksson C, Glosli H, Iversen N, Sullivan GJ, Sandset PM, Stavik B, Chollet ME. Gene editing in patient derived 3D liver organoids to address severe coagulation factor VII deficiency. Haematologica, 2025;111:941-954.
2) Harrison Sean P, Siller R, Tanaka Y, Chollet, ME, de la Morena-Barrio, ME, Xiang Y, Patterson B, Andersen E, Bravo-Pérez C, Kempf H, Åsrud KS, Lunov O, Dejneka A, Mowinckel M-C, Stavik B, Sandset PM, Melum E, Baumgarten S, Bonanini F, Kurek D, Mathapati S, Almaas R, Sharma K, Wilson SR, Skottvoll FS, Boger IC, Bogen IL, Nyman TA, Wu JJ, Bezrouk A, Cizkova D, Corral J, Mokry J, Zweigerdt R, Park I-H, Sullivan GJ. Scalable production of tissue-like vascularized liver organoids from human PSCs. Experimental and Molecular Medicine. 2023;55(9):2005-2024.
3) Roman G, Stavik B, Lauritzen KH, Sandset PM, Harrison SP, Sullivan GJ and Chollet ME. iPSC-derived liver organoids and inherited bleeding disorders: Potential and future perspectives. Frontiers in Physiology. 2023;14:1094249.