Deciphering & Drugging DNA Repair 

The Deciphering & drugging DNA Repair (DDR) lab aims at deciphering how human cells respond to DNA damage to identify and validate new druggable complexes for cancer treatment. Our landmark discoveries include alternative End-Joining (2004), mechanisms antagonizing Ku association to some DNA ends (2016, 2020), a new function for BRCA1 at centromeres (2014, 2021), the characterization of Ku interactions with DNA repair factors (2018, 2023) and the discovery of the mechanism of action of several bioactive small molecules (2017, 2021, 2022).

Through a multidisciplinary approach, we tackle an important problem: how DNA repair works and how can we modulate it through the use of small molecules.

DNA repair controls the outcome of several anticancer therapies. As such there is a huge interest for novel small molecules modulating DNA repair. In the DDR lab, we decipher how cells respond to DNA damage, more specifically to the most harmful type, DNA Double-Strand Breaks (DSBs). We use the resulting knowledge to design and perform targeted and phenotypical based screens to identify new modulators of specific aspects of the DNA damage response. To achieve our goals, we rely on high- and super-resolution imaging, molecular and cell biology, genomics, small molecules screening, chemical probes and on multiple collaborations with structural biologists and chemists. We recently implemented in the lab several complementary approaches to decipher how biologically active small molecules act (Bombarde et al. Mol Cancer Ther 2017; Bossaert, Pipier et al. 2021; Demange, Joly, Marcoux et al. eLife 2022).

The interplay between DNA repair mechanisms

Two principal DSB repair mechanisms co-exist in human cells: Non- Homologous End Joining (NHEJ) and Homologous Recombination (HR). Using a new method for imaging NHEJ proteins, we recently discovered the main mechanism antagonizing NHEJ proteins association to special DSBs, thereby allowing HR to proceed. We established that these mechanisms are druggable and that their inhibition triggers toxic DNA repair events in response to some anticancer agents (Britton et al. 2013 J Cell Biol; Chanut, Britton et al. 2016 Nat Commun; Britton, Chanut et al. 2020 Nucleic Acids Res).

How DNA repair proteins associate on damaged chromatin

We recently characterized how the repair factors APLF and XLF associate with the NHEJ core protein Ku at DNA damage, thereby characterizing two new interaction interfaces at the functional and structural levels (Nemoz, Ropars, Frit et al. 2018 Nat Struct Mol Biol; Seif-El-Dahan, Kefala-Stavridi, Frit, Hardwick et al. 2023). We also investigate how the genomic context, such as heterochromatin and transcription, affect DNA repair and genome stability. Using laser micro-irradiation and live imaging, we discovered a novel mechanism promoting the exclusion of a large number of RNA-binding proteins from DNA damage sites, thereby preventing the local accumulation of R-loops (Britton, Dernoncourt et al. Nucleic Acids Res 2014).

How stabilizing specific DNA structures triggers DNA damage

Small molecules stabilizing secondary DNA structures, such as G-quadruplexes (G4) ligands, trigger DNA damage (Zell et al. 2020 RSC Chem Biol). Through an unbiased genomic approach, we recently discovered that DNA topoisomerase 2 alpha is responsible for the production of DSB by several G4 ligands (Bossaert, Pipier et al. 2021 eLife), including CX-5461 a molecule undergoing clinical trials in oncology. We also contributed to develop novel ligands of another secondary structure, the Three-way Junction, and established that they also trigger DNA damage by a mechanism that we are currently investigating (Duskova 2019 J Med Chem; Duskova et al. 2020 J Am Chem Soc; Zell et al. Nucleic Acids Res 2021).

Team members

Research Scientists

Nadia Barboule (CNRS)
Sébastien Britton (CNRS)
Patrick Calsou (Inserm)
Philippe Frit (CNRS)
Dennis Gomez (CNRS)
Florence Larminat (CNRS)
Marie-Jeanne Pillaire (Inserm)

Research Engineers

Maëlle Caroff
Antonio Peixoto (CNRS)
Nadège Preuilh
Carine Racca (CNRS)
Maria Fidelia Susanto

Postdoctoral Fellow

Léa Marie

PhD Students

Madeleine Bossaert
Margaux Bossuat
Jeanne Chauvat
Maryam Salim

Our research projects

Demange, Joly, Marcoux, et al (2022) SDR enzymes oxidize specific lipidic alkynylcarbinols into cytotoxic protein-reactive species. eLife

Bossaert, Pipier, et al (2021) Transcription-associated topoisomerase 2alpha (TOP2A) activity is a major effector of cytotoxicity induced by G-quadruplex ligands. eLife

Racca et al (2021) BRCA1 prevents R-loop-associated centromeric instability. Cell death & disease

Britton, Chanut et al (2020) ATM antagonizes NHEJ proteins assembly and DNA-ends synapsis at single-ended DNA double strand breaks. Nucleic Acids Res 

Zell et al (2021) Dual targeting of higher-order DNA structures by azacryptands induces DNA junction-mediated DNA damage in cancer cells. Nucleic Acids Res

Nemoz, Ropars, Frit et al (2018) XLF and APLF bind Ku80 at two remote sites to ensure DNA repair by non-homologous end joining. Nat Struct Mol Biol

High-resolution imaging of DNA Double-Strand Breaks (DSBs) in human cells treated with ionizing radiations in presence of a DDR inhibitor. Red: DSBs visualized with gammaH2AX; Green: Actin cytoskeleton; Blue: DNA.
© Sébastien Britton.