Project XPF-ERCC1

XPF-ERCC1 – Structural and kinetic studies of XPF/ERCC1-DNA complex for drug discovery

ABSTRACT

Cancer chemotherapeutic drugs, such as platinum agents, work by producing DNA damage that causes cell-cycle arrest and cell death. However, DNA repair pathways can enable tumor cells to survive DNA damage induced by chemotherapeutic treatments. Platinum resistance is a major clinical problem in cancer treatment and there is a considerable interest in inhibitors of DNA repair pathways that can increase efficiency of chemotherapy, when administered together.
An attractive target to circumvent platinum resistance is XPF/ERCC1 heterodimer. XPF/ERCC1 is a structure specific endonuclease that cleaves double- to single-stranded DNA junctions, and its function is directly linked to repair of DNA cross-links induced by platinum agents. Here, we propose a multi-disciplinary methodology that combines structural studies by Nuclear Magnetic Resonance Spectroscopy with kinetic studies by Kinetic Capillary Electrophoresis to elucidate the mechanisms of XPF/ERCC1 function and modulate its activity for potential therapeutic benefit. Structural and molecular insights of DNA binding will be coupled with kinetic rates of DNA cleavage and enzyme/substrate interactions. The combined information will be used to develop XPF/ERCC1 inhibitors by in silico screening.
Currently structure-kinetics relationship is not well explored in drug discovery, although the residence lifetime of a drug determines the duration of a pharmacological effect. The ability to combine structural and kinetic parameters in validating computational hits should identify compounds with desired pharmacological properties at early stages and facilitate faster optimization phase. The research proposed here is highly beneficial to public health, because it would allow to develop combination therapies for cancer treatment.

PROJECT AIMS

1. Understanding how DNA binding properties of XPF/ERCC1 heterodimer regulate catalytic activity.
2. Design and evaluation of small molecule inhibitors to modulate XPF/ERCC1 activity.

OBJECTIVES FOR TRANSFER OF KNOWLEDGE

1. Unique expertise in kinetic analysis of protein-DNA and protein-protein interactions and understanding of research methodology.
2. Participation in group discussions and meetings with other researchers and students at CEITEC.
3. Dissemination of obtained results through publications in peer-reviewed journals, presentation at international conferences in the form of posters and lectures, internal seminars, and discussion sessions with students and colleagues.
4. Participation in and organizing of outreach activities, such as CEITEC Open Day, Workshop day, Publications in popular-science magazines, Research-related communications resources etc.