Réparation, Radiations et Thérapies innovantes anticancer

Publications de l’équipe

Année de publication : 2014

L Sancey, F Lux, S Kotb, S Roux, S Dufort, A Bianchi, Y Crémillieux, P Fries, J-L Coll, C Rodriguez-Lafrasse, M Janier, M Dutreix, M Barberi-Heyob, F Boschetti, F Denat, C Louis, E Porcel, S Lacombe, G Le Duc, E Deutsch, J-L Perfettini, A Detappe, C Verry, R Berbeco, K T Butterworth, S J McMahon, K M Prise, P Perriat, O Tillement (2014 Jul 4)

The use of theranostic gadolinium-based nanoprobes to improve radiotherapy efficacy.

The British journal of radiology : 20140134 : DOI : 10.1259/bjr.20140134 En savoir plus
Résumé

A new efficient type of gadolinium-based theranostic agent (AGuIX®) has recently been developed for MRI-guided radiotherapy (RT). These new particles consist of a polysiloxane network surrounded by a number of gadolinium chelates, usually 10. Owing to their small size (<5 nm), AGuIX typically exhibit biodistributions that are almost ideal for diagnostic and therapeutic purposes. For example, although a significant proportion of these particles accumulate in tumours, the remainder is rapidly eliminated by the renal route. In addition, in the absence of irradiation, the nanoparticles are well tolerated even at very high dose (10 times more than the dose used for mouse treatment). AGuIX particles have been proven to act as efficient radiosensitizers in a large variety of experimental in vitro scenarios, including different radioresistant cell lines, irradiation energies and radiation sources (sensitizing enhancement ratio ranging from 1.1 to 2.5). Pre-clinical studies have also demonstrated the impact of these particles on different heterotopic and orthotopic tumours, with both intratumoural or intravenous injection routes. A significant therapeutical effect has been observed in all contexts. Furthermore, MRI monitoring was proven to efficiently aid in determining a RT protocol and assessing tumour evolution following treatment. The usual theoretical models, based on energy attenuation and macroscopic dose enhancement, cannot account for all the results that have been obtained. Only theoretical models, which take into account the Auger electron cascades that occur between the different atoms constituting the particle and the related high radical concentrations in the vicinity of the particle, provide an explanation for the complex cell damage and death observed.

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Flavien Devun, Julian Biau, Michel Huerre, Amélie Croset, Jian-Sheng Sun, Alban Denys, Marie Dutreix (2014 Apr 14)

Colorectal cancer metastasis: the DNA repair inhibitor Dbait increases sensitivity to hyperthermia and improves efficacy of radiofrequency ablation.

Radiology : 736-46 : DOI : 10.1148/radiol.13130805 En savoir plus
Résumé

To assess the usefulness of combining hyperthermia with a DNA repair inhibitor (double-strand break bait [Dbait]) and its potential application to radiofrequency ablation (RFA) in a preclinical model of human colorectal cancer.

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Imen Miladi, Christophe Alric, Sandrine Dufort, Pierre Mowat, Aurélie Dutour, Céline Mandon, Gautier Laurent, Elke Bräuer-Krisch, Nirmitha Herath, Jean-Luc Coll, Marie Dutreix, François Lux, Rana Bazzi, Claire Billotey, Marc Janier, Pascal Perriat, Géraldine Le Duc, Stéphane Roux, Olivier Tillement (2014 Mar 25)

The in vivo radiosensitizing effect of gold nanoparticles based MRI contrast agents.

Small (Weinheim an der Bergstrasse, Germany) : 1116-24 : DOI : 10.1002/smll.201470036 En savoir plus
Résumé

Owing to the high atomic number (Z) of gold element, the gold nanoparticles appear as very promising radiosensitizing agents. This character can be exploited for improving the selectivity of radiotherapy. However, such an improvement is possible only if irradiation is performed when the gold content is high in the tumor and low in the surrounding healthy tissue. As a result, the beneficial action of irradiation (the eradication of the tumor) should occur while the deleterious side effects of radiotherapy should be limited by sparing the healthy tissue. The location of the radiosensitizers is therefore required to initiate the radiotherapy. Designing gold nanoparticles for monitoring their distribution by magnetic resonance imaging (MRI) is an asset due to the high resolution of MRI which permits the accurate location of particles and therefore the determination of the optimal time for the irradiation. We recently demonstrated that ultrasmall gold nanoparticles coated by gadolinium chelates (Au@DTDTPA-Gd) can be followed up by MRI after intravenous injection. Herein, Au@DTDTPA and Au@DTDTPA-Gd were prepared in order to evaluate their potential for radiosensitization. Comet assays and in vivo experiments suggest that these particles appear well suited for improving the selectivity of the radiotherapy. The dose which is used for inducing similar levels of DNA alteration is divided by two when cells are incubated with the gold nanoparticles prior to the irradiation. Moreover, the increase in the lifespan of tumor bearing rats is more important when the irradiation is performed after the injection of the gold nanoparticles. In the case of treatment of rats with a brain tumor (9L gliosarcoma, a radio-resistant tumor in a radiosensitive organ), the delay between the intravenous injection and the irradiation was determined by MRI.

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Année de publication : 2013

Ewa Kotula, Wolfgang Faigle, Nathalie Berthault, Florent Dingli, Damarys Loew, Jian-Sheng Sun, Marie Dutreix, Maria Quanz (2013 Nov 28)

DNA-PK target identification reveals novel links between DNA repair signaling and cytoskeletal regulation.

PloS one : e80313 : DOI : 10.1371/journal.pone.0080313 En savoir plus
Résumé

The DNA-dependent protein kinase (DNA-PK) may function as a key signaling kinase in various cellular pathways other than DNA repair. Using a two-dimensional gel electrophoresis approach and stable DNA double-strand break-mimicking molecules (Dbait32Hc) to activate DNA-PK in the nucleus and cytoplasm, we identified 26 proteins that were highly phosphorylated following DNA-PK activation. Most of these proteins are involved in protein stability and degradation, cell signaling and the cytoskeleton. We investigated the relationship between DNA-PK and the cytoskeleton and found that the intermediate filament (IF) vimentin was a target of DNA-PK in vitro and in cells. Vimentin was phosphorylated at Ser459, by DNA-PK, in cells transfected with Dbait32Hc. We produced specific antibodies and showed that Ser459-P-vimentin was mostly located at cell protrusions. In migratory cells, the vimentin phosphorylation induced by Dbait32Hc was associated with a lower cellular adhesion and migration capacity. Thus, this approach led to the identification of downstream cytoplasmic targets of DNA-PK and revealed a connection between DNA damage signaling and the cytoskeleton.

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Amelie Croset, Fabrice P Cordelières, Nathalie Berthault, Cyril Buhler, Jian-Sheng Sun, Maria Quanz, Marie Dutreix (2013 Jun 12)

Inhibition of DNA damage repair by artificial activation of PARP with siDNA.

Nucleic acids research : 7344-55 : DOI : 10.1093/nar/gkt522 En savoir plus
Résumé

One of the major early steps of repair is the recruitment of repair proteins at the damage site, and this is coordinated by a cascade of modifications controlled by phosphatidylinositol 3-kinase-related kinases and/or poly (ADP-ribose) polymerase (PARP). We used short interfering DNA molecules mimicking double-strand breaks (called Dbait) or single-strand breaks (called Pbait) to promote DNA-dependent protein kinase (DNA-PK) and PARP activation. Dbait bound and induced both PARP and DNA-PK activities, whereas Pbait acts only on PARP. Therefore, comparative study of the two molecules allows analysis of the respective roles of the two signaling pathways: both recruit proteins involved in single-strand break repair (PARP, XRCC1 and PCNA) and prevent their recruitment at chromosomal damage. Dbait, but not Pbait, also inhibits recruitment of proteins involved in double-strand break repair (53BP1, NBS1, RAD51 and DNA-PK). By these ways, Pbait and Dbait disorganize DNA repair, thereby sensitizing cells to various treatments. Single-strand breaks repair inhibition depends on direct trapping of the main proteins on both molecules. Double-strand breaks repair inhibition may be indirect, resulting from the phosphorylation of double-strand breaks repair proteins and chromatin targets by activated DNA-PK. The DNA repair inhibition by both molecules is confirmed by their synthetic lethality with BRCA mutations.

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Francesca Farina, Paolo Pierobon, Cédric Delevoye, Jordan Monnet, Florent Dingli, Damarys Loew, Maria Quanz, Marie Dutreix, Giovanni Cappello (2013 Apr 2)

Kinesin KIFC1 actively transports bare double-stranded DNA.

Nucleic acids research : 4926-37 : DOI : 10.1093/nar/gkt204 En savoir plus
Résumé

During the past years, exogenous DNA molecules have been used in gene and molecular therapy. At present, it is not known how these DNA molecules reach the cell nucleus. We used an in cell single-molecule approach to observe the motion of exogenous short DNA molecules in the cytoplasm of eukaryotic cells. Our observations suggest an active transport of the DNA along the cytoskeleton filaments. We used an in vitro motility assay, in which the motion of single-DNA molecules along cytoskeleton filaments in cell extracts is monitored; we demonstrate that microtubule-associated motors are involved in this transport. Precipitation of DNA-bound proteins and mass spectrometry analyses reveal the preferential binding of the kinesin KIFC1 on DNA. Cell extract depletion of kinesin KIFC1 significantly decreases DNA motion, confirming the active implication of this molecular motor in the intracellular DNA transport.

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Année de publication : 2012

Marie Dutreix (2012 Jul 26)

Safety control for apoptotic irreversibility.

Proceedings of the National Academy of Sciences of the United States of America : 12844-5 : DOI : 10.1073/pnas.1210736109 En savoir plus
Résumé

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Nicolas Coquery, Nicolas Pannetier, Régine Farion, Aurélie Herbette, Leire Azurmendi, Didier Clarencon, Stéphane Bauge, Véronique Josserand, Claire Rome, Jean-Luc Coll, Jian-Sheng Sun, Emmanuel L Barbier, Marie Dutreix, Chantal C Remy (2012 Jul 21)

Distribution and radiosensitizing effect of cholesterol-coupled Dbait molecule in rat model of glioblastoma.

PloS one : e40567 : DOI : 10.1371/journal.pone.0040567 En savoir plus
Résumé

Glioma is the most aggressive tumor of the brain and the most efficient treatments are based on radiotherapy. However, tumors are often resistant to radiotherapy due to an enhanced DNA repair activity. Short and stabilized DNA molecules (Dbait) have recently been proposed as an efficient strategy to inhibit DNA repair in tumor.

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Maria Quanz, Aurélie Herbette, Mano Sayarath, Leanne de Koning, Thierry Dubois, Jian-Sheng Sun, Marie Dutreix (2012 Jan 23)

Heat shock protein 90α (Hsp90α) is phosphorylated in response to DNA damage and accumulates in repair foci.

The Journal of biological chemistry : 8803-15 : DOI : 10.1074/jbc.M111.320887 En savoir plus
Résumé

DNA damage triggers a complex signaling cascade involving a multitude of phosphorylation events. We found that the threonine 7 (Thr-7) residue of heat shock protein 90α (Hsp90α) was phosphorylated immediately after DNA damage. The phosphorylated Hsp90α then accumulated at sites of DNA double strand breaks and formed repair foci with slow kinetics, matching the repair kinetics of complex DNA damage. The phosphorylation of Hsp90α was dependent on phosphatidylinositol 3-kinase-like kinases, including the DNA-dependent protein kinase (DNA-PK) in particular. DNA-PK plays an essential role in the repair of DNA double strand breaks by nonhomologous end-joining and in the signaling of DNA damage. It is also present in the cytoplasm of the cell and has been suggested to play a role in cytoplasmic signaling pathways. Using stabilized double-stranded DNA molecules to activate DNA-PK, we showed that an active DNA-PK complex could be assembled in the cytoplasm, resulting in phosphorylation of the cytoplasmic pool of Hsp90α. In vivo, reverse phase protein array data for tumors revealed that basal levels of Thr-7-phosphorylated Hsp90α were correlated with phosphorylated histone H2AX levels. The Thr-7 phosphorylation of the ubiquitously produced and secreted Hsp90α may therefore serve as a surrogate biomarker of DNA damage. These findings shed light on the interplay between central DNA repair enzymes and an essential molecular chaperone.

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Année de publication : 2011

P Mowat, A Mignot, W Rima, F Lux, O Tillement, C Roulin, M Dutreix, D Bechet, S Huger, L Humbert, M Barberi-Heyob, M T Aloy, E Armandy, C Rodriguez-Lafrasse, G Le Duc, S Roux, P Perriat (2011 Nov 22)

In vitro radiosensitizing effects of ultrasmall gadolinium based particles on tumour cells.

Journal of nanoscience and nanotechnology : 7833-9 En savoir plus
Résumé

Since radiotherapy is widely used in cancer treatment, it is essential to develop strategies which lower the irradiation burden while increasing efficacy and become efficient even in radio resistant tumors. Our new strategy is relying on the development of solid hybrid nanoparticles based on rare-earth such as gadolinium. In this paper, we then evidenced that gadolinium-based particles can be designed to enter efficiently into the human glioblastoma cell line U87 in quantities that can be tuned by modifying the incubation conditions. These sub-5 nm particles consist in a core of gadolinium oxide, a shell of polysiloxane and are functionalized by diethylenetriaminepentaacetic acid (DTPA). Although photoelectric effect is maximal in the [10-100 keV] range, such particles were found to possess efficient in-vitro radiosensitizing properties at an energy of 660 keV by using the « single-cell gel electrophoresis comet assay, » an assay that measures the number of DNA damage that occurs during irradiation. Even more interesting, the particles have been evidenced by MTT assays to be also efficient radiosensitizers at an energy of 6 MeV for doses comprised between 2 and 8 Gy. The properties of the gadolinium-based particles give promising opening to a particle-assisted radio-therapy by using irradiation systems already installed in the majority of hospitals.

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Wiebke Solass, Aurélie Herbette, Tina Schwarz, Alexander Hetzel, Jian-Sheng Sun, Marie Dutreix, Marc A Reymond (2011 Nov 2)

Therapeutic approach of human peritoneal carcinomatosis with Dbait in combination with capnoperitoneum: proof of concept.

Surgical endoscopy : 847-52 : DOI : 10.1007/s00464-011-1964-y En savoir plus
Résumé

Peritoneal carcinomatosis is an unmet medical need. Laparoscopy offers a unique opportunity to control and to steer the operating environment during surgery by loading carbon dioxide with a therapeutic substance and creating the so-called therapeutic capnoperitoneum. We have treated a human sample of peritoneal carcinomatosis from an endometrial adenocarcinoma ex vivo just after surgery.

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Flavie Luciani, Delphine Champeval, Aurélie Herbette, Laurence Denat, Bouchra Aylaj, Silvia Martinozzi, Robert Ballotti, Rolf Kemler, Colin R Goding, Florian De Vuyst, Lionel Larue, Véronique Delmas (2011 Aug 25)

Biological and mathematical modeling of melanocyte development.

Development (Cambridge, England) : 3943-54 : DOI : 10.1242/dev.067447 En savoir plus
Résumé

We aim to evaluate environmental and genetic effects on the expansion/proliferation of committed single cells during embryonic development, using melanoblasts as a paradigm to model this phenomenon. Melanoblasts are a specific type of cell that display extensive cellular proliferation during development. However, the events controlling melanoblast expansion are still poorly understood due to insufficient knowledge concerning their number and distribution in the various skin compartments. We show that melanoblast expansion is tightly controlled both spatially and temporally, with little variation between embryos. We established a mathematical model reflecting the main cellular mechanisms involved in melanoblast expansion, including proliferation and migration from the dermis to epidermis. In association with biological information, the model allows the calculation of doubling times for melanoblasts, revealing that dermal and epidermal melanoblasts have short but different doubling times. Moreover, the number of trunk founder melanoblasts at E8.5 was estimated to be 16, a population impossible to count by classical biological approaches. We also assessed the importance of the genetic background by studying gain- and loss-of-function β-catenin mutants in the melanocyte lineage. We found that any alteration of β-catenin activity, whether positive or negative, reduced both dermal and epidermal melanoblast proliferation. Finally, we determined that the pool of dermal melanoblasts remains constant in wild-type and mutant embryos during development, implying that specific control mechanisms associated with cell division ensure half of the cells at each cell division to migrate from the dermis to the epidermis. Modeling melanoblast expansion revealed novel links between cell division, cell localization within the embryo and appropriate feedback control through β-catenin.

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N Berthault, B Maury, C Agrario, A Herbette, J-S Sun, N Peyrieras, M Dutreix (2011 Jul 30)

Comparison of distribution and activity of nanoparticles with short interfering DNA (Dbait) in various living systems.

Cancer gene therapy : 695-706 : DOI : 10.1038/cgt.2011.39 En savoir plus
Résumé

Introducing small DNA molecules (Dbait) impairs the repair of damaged chromosomes and provides a new method for enhancing the efficiency of radiotherapy in radio-resistant tumors. The radiosensitizing activity is dependent upon the efficient delivery of Dbait molecules into the tumor cells. Different strategies have been compared, to improve this key step. We developed a pipeline of assays to select the most efficient nanoparticles and administration protocols before preclinical assays: (i) molecular analyses of complexes formed with Dbait molecules, (ii) cellular tests for Dbait uptake and activity, (iii) live zebrafish embryo confocal microscopy monitoring for in vivo distribution and biological activity of the nanoparticles and (iv) tumor growth and survival measurement on mice with xenografted tumors. Two classes of nanoparticles were compared, polycationic polymers with linear or branched polyethylenimine (PEI) and covalently attached cholesterol (coDbait). The most efficient Dbait transfection was observed with linear PEI complexes, in vitro and in vivo. Doses of coDbait ten-fold higher than PEI/Dbait nanoparticles, and pretreatment with chloroquine, were required to obtain the same antitumoral effect on xenografted melanoma. However, with a 22-fold lower ‘efficacy dose/toxicity dose’ ratio as compared with Dbait/PEI, coDbait was selected for clinical trials.

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Judith Miné-Hattab, Geneviève Fleury, Chantal Prevost, Marie Dutreix, Jean-Louis Viovy (2011 Apr 13)

Optimizing the design of oligonucleotides for homology directed gene targeting.

PloS one : e14795 : DOI : 10.1371/journal.pone.0014795 En savoir plus
Résumé

Gene targeting depends on the ability of cells to use homologous recombination to integrate exogenous DNA into their own genome. A robust mechanistic model of homologous recombination is necessary to fully exploit gene targeting for therapeutic benefit.

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Flavien Devun, Guilhem Bousquet, Julian Biau, Aurélie Herbette, Christophe Roulin, Frédérique Berger, Jian-Sheng Sun, Sylvie Robine, Marie Dutreix (2011 Apr 10)

Preclinical study of the DNA repair inhibitor Dbait in combination with chemotherapy in colorectal cancer.

Journal of gastroenterology : 266-75 : DOI : 10.1007/s00535-011-0483-x En savoir plus
Résumé

Dbait molecules are a new class of DNA repair inhibitors triggering false DNA damage signaling in cancer cells. Dbait has already been shown to be effective in combination with radiotherapy. The aim of this study was to assess the adjuvant impact of Dbait on chemotherapy in vitro and in mouse models of colorectal cancer.

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