Overview

Supervisor

Martin Gregor

Project description

Hepatocellular carcinoma (HCC) is an aggressive liver malignancy with limited therapeutic options. Tumor progression critically depends on cytoskeletal organization and mechanotransduction, processes in which the cytolinker protein plectin plays a central role. Plectin integrates intermediate filaments, actin filaments, and microtubules and has emerged as a promising therapeutic target in HCC.

This PhD project aims to elucidate the molecular and cellular mechanisms of plectin inhibition using plecstatin, a first-in-class high-affinity plectin inhibitor. The project combines structural characterization with functional analyses in clinically relevant human tumor models.

In the first part, the student will define the plecstatin binding site on the plectin molecule. This will involve in vitro expression and purification of selected plectin domains and engineered variants, followed by biophysical and structural analyses. Hydrogen-deuterium exchange mass spectrometry will be used to identify plecstatin-induced conformational changes and interaction interfaces. Selected plectin–plecstatin complexes will be further analyzed by cryo-electron microscopy to resolve the structural basis of inhibitor binding and its impact on plectin function.

In the second part, the student will investigate plecstatin’s mode of action in patient-derived HCC explants maintained ex vivo to preserve native tumor architecture. Using advanced microscopy approaches, the project will assess plecstatin-induced changes in cytoskeletal organization, cell mechanics, and tumor cell behavior. These analyses will be complemented by quantitative proteomics to identify plecstatin-dependent remodeling of cytoskeletal and signaling networks in human tumor tissue.

By integrating structural biology with functional analyses in patient-derived HCC models, this project will provide mechanistic insight into cytoskeletal targeting strategies in cancer and support the development of plectin-directed therapies.

Candidate profile

Master’s degree in molecular biology, cell biology, biochemistry, biophysics, or a related discipline
Strong interest in cancer biology, cytoskeleton, or structural biology
High motivation, independence, and ability to work in a multidisciplinary research environment
Excellent organizational skills and ability to plan and prioritize experimental work
Strong written and verbal communication skills in English
Prior experience with protein expression and purification, mass spectrometry, advanced microscopy, or work with primary human tissues will be considered an advantage but is not required. The candidate is expected to actively contribute to data analysis, manuscript preparation, and international collaborations.

Suggested reading

Outla Z, Oyman-Eyrilmez G, Korelova K, Prechova M, Frick L, Sarnova L, Bisht P, Novotna P, Kosla J, Bortel P, Borutzki Y, Bileck A, Gerner C, Rahbari M, Rahbari N, Birgin E, Kvasnicova B, Galisova A, Sulkova K, Bauer A, Jobe N, Tolde O, Sticova E, Rösel D, O’Connor T, Otahal M, Jirak D, Heikenwälder M, Wiche G, Meier-Menches SM, Gregor M: Plectin-mediated cytoskeletal crosstalk as a target for inhibition of hepatocellular carcinoma growth and metastasis. Elife 2025 13:RP102205.
Prechova M, Korelova K, Gregor M: Plectin. Curr Biol 2023 33(4): R128-R130.
Meier S. M., Kreutz D., Winter L., Klose M. H. M., Cseh K., Weiss T., Bileck A., Alte B., Mader J. C., Jana S., Chatterjee A., Bhattacharyya A., Hejl M., Jakupec M. A., Heffeter P., Berger W., Hartinger C. G., Keppler B. K., Wiche G., Gerner C.: An organoruthenium anticancer agent shows unexpected target selectivity for plectin. Angew Chem Int Ed Engl 56(28):8267-8271.
Prechova M, Adamova Z, Schweizer AL, Maninova M, Bauer A, Kah D, Meier-Menches SM, Wiche G, Fabry B, Gregor M: Plectin-mediated cytoskeletal crosstalk controls cell tension and cohesion in epithelial sheets. J Cell Biol 2022 221(3):e202105146.

APPLY HERE: https://www.img.cas.cz/phd