Department Molecular Genetics
Principal investigator Hannes Lans
E-mail address w.lans@erasmusmc.nl
Website https://lanslab.eu/
Overcoming pancreatic ductal adenocarcinoma chemoresistance by targeting DNA repair
Supervisor: Romy Paap, r.paap@erasmusmc.nl
Resistance to the widely used anti-cancer drug FOLFIRINOX is a prominent issue in pancreatic ductal adenocarcinoma (PDAC) treatment. FOLFIRINOX derives its cytotoxicity from its ability to induce DNA damage; however, the DNA repair deficiencies found in many pancreatic tumors are often not investigated. Using patient derived PDAC organoids, individual tumor therapy responses can be analyzed both functionally and molecularly. In this project, we will test and develop DNA repair-targeting therapies, together with FOLFIRINOX, to overcome PDAC chemoresistance. We will use different genomic analyses, genetics, and functional testing in PDAC organoids to analyze DNA repair capacity of individual PDAC tumors and genetically and chemically inhibit various DNA damage response (DDR) pathways to investigate therapeutic opportunities for tailored treatment.
Techniques
- Cell/organoid culture
- Survival assays
- DNA repair assays
- Microscopy
Further reading
Organoids Derived from Neoadjuvant FOLFIRINOX Patients Recapitulate Therapy Resistance in Pancreatic Ductal Adenocarcinoma. Farshadi et al. Clin Cancer Res. 2021
Detection of oxaliplatin- and cisplatin-DNA lesions requires different global genome repair mechanisms that affect their clinical efficacy. Slyskova et al. NAR Cancer. 2023
Understanding nucleotide excision repair and its roles in cancer and ageing. Marteijn et al. Nat Rev Mol Cell Biol. 2014
Deciphering Disease Mechanisms in TFIIH Mutations: From DNA Repair to Clinical Symptoms
Supervisor: David Häckes/Hannes Lans, d.hackes@erasmusmc.nl/w.lans@erasmusmc.nl
This project investigates the molecular mechanisms behind diverse diseases caused by mutations in TFIIH, a critical protein complex in DNA repair and transcription initiation. Our goal is to understand how specific TFIIH defects lead to disorders such as Xeroderma pigmentosum, Cockayne syndrome, and Trichothiodystrophy. Using advanced imaging techniques, we examine TFIIH function in human cells and explore its impact on DNA repair and transcription. Additionally, C. elegans serves as a model to study TFIIH’s role in differentiated tissues, with a focus on neuronal degradation. This research offers students hands-on experience in molecular biology, using a combination of innovative methodologies.
Techniques
- Various imaging-based assays
- Functional assays to evaluate DNA repair efficiency
- C. elegans as a model organism
Further reading
Arjan F. Theil, David Häckes, Hannes Lans TFIIH central activity in nucleotide excision repair to prevent disease. DNA Repair 132, (2024). https://doi.org/10.1016/j.dnarep.2023.103568
Muniesa-Vargas, A., Davó-Martínez, C., Ribeiro-Silva, C. et al. Persistent TFIIH binding to non-excised DNA damage causes cell and developmental failure. Nat Commun 15, 3490 (2024). https://doi.org/10.1038/s41467-024-47935-9
Genome maintenance mechanisms that control ERCC1-XPF endonuclease activity
Supervisor: Ulkem Usla Kaynak, u.kaynak@erasmusmc.nl
The endonuclease ERCC1-XPF protects cancer cells against DNA damaging chemotherapeutic agents due to its vital role in nucleotide excision repair (NER) and inter-strand crosslink repair (ICLR). The regulation of ERCC1-XPF in NER is known; however, how ERCC1-XPF activity is regulated in ICLR is not well understood, especially in non-replicating cells. To study ERCC1-XPF function and identify the proteins and mechanisms that control its activity in replication-independent ICLR, we developed a new imaging platform, with which we can monitor the localization of ERCC1-XPF to UVA laser-induced psoralen-DNA crosslinks in living cells. We also perform immunoprecipitation to determine the interactors of ERCC1-XPF during ICLR. Through our studies, we hope to unravel the exact mechanism by which ICLs are repaired by ERCC1-XPF.
Techniques
- Cell culture
- Transfection
- Confocal microscopy
- Immunoblotting
- Immunoprecipitation
- DNA repair assays
Further reading
Sabatella, M., Pines, A., Slyskova, J., Vermeulen, W., & Lans, H. (2020). ERCC1-XPF targeting to psoralen-DNA crosslinks depends on XPA and FANCD2. Cellular and molecular life sciences : CMLS, 77(10), 2005–2016. https://doi.org/10.1007/s00018-019-03264-5
Muniesa-Vargas, A., Davó-Martínez, C., Ribeiro-Silva, C., van der Woude, M., Thijssen, K. L., Haspels, B., Häckes, D., Kaynak, Ü. U., Kanaar, R., Marteijn, J. A., Theil, A. F., Kuijten, M. M. P., Vermeulen, W., & Lans, H. (2024). Persistent TFIIH binding to non-excised DNA damage causes cell and developmental failure. Nature communications, 15(1), 3490. https://doi.org/10.1038/s41467-024-47935-9