UMR9187 / U1196 – Chimie et Modélisation pour la Biologie du Cancer (CMBC)

Publications de l’unité

Année de publication : 2018

Chen, Su; Poyer, Florent; Garcia, Guillaume; Fiorini-Debuisschert, Céline; Rosilio, Véronique and Maillard, Philippe (2018 Feb 14)

Amphiphilic Glycoconjugated Porphyrin Heterodimers as Two-Photon Excitable Photosensitizers: Design, Synthesis, Photophysical and Photobiological Studies

ChemistrySelect : 3 : 1887-1897 : DOI : 10.1002/slct.201703013 En savoir plus
Résumé

A new family of amphiphilic glycoconjugated porphyrin dimers usable in two-photon photodynamic therapy was designed, prepared and characterized. The physicochemical and biological data were presented and discussed. Their two-photon absorption cross section was determined, revealing quite good performances of the compounds (δ ≈ 300 GM at 880 nm for 2 and 3 and δ ≈ 1000 GM at 840 nm for 5 and 6). Despite their promising photophysical properties, porphyrin dimers are penalized by their low solubility and an insufficient penetration into tumour cells. Incorporation into nanocarriers such as lipid vesicles could be profitable, provided that the positioning and degree of freedom of the saccharide moieties are optimal to target the receptors of the cells of interest.

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Marius Mamone, Jessy Aziz, Julie Le Bescont, Sandrine Piguel (2018 Jan 18)

Aminocarbonylation of N-Containing Heterocycles with Aromatic Amines Using Mo(CO)6

Synthesis : 50 : 1521-1526 : DOI : 10.1055/s-0037-1609152 En savoir plus
Résumé

We describe herein the palladium-catalyzed aminocarbonylation of nitrogen-containing heterocycles with aniline derivatives using molybdenum hexacarbonyl as a CO solid source, expanding the scope of the limited examples. This method is compatible with a variety of substitutions on the aniline moiety. The simple reaction conditions include easily available Pd(dppf)Cl2 catalyst, DBU as base in DMF at 120 °C for 3 hours in sealed tube thereby leading to the isolation of 21 compounds with yields ranging from 18 to 82%. We also show that double aminocarbonylation reactions are possible in satisfactory yields regarding both coupling partners.

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Hammerer F., Poyer F., Fourmois L., Chen S., Garcia G., Teulade-Fichou M.P., Maillard P., Mahuteau-Betzer F. (2018 Jan 1)

Mitochondria-targeted cationic porphyrin-triphenylamine hybrids for enhanced two-photon photodynamic therapy

Bioorganic & Medicinal Chemistry : 26 : 107-118 : DOI : 10.1016/j.bmc.2017.11.024 En savoir plus
Résumé

The proof of concept for two-photon activated photodynamic therapy has already been achieved for cancer treatment but the efficiency of this approach still heavily relies on the availability of photosensitizers combining high two-photon absorption and biocompatibility. In this line we recently reported on a series of porphyrin-triphenylamine hybrids which exhibit high singlet oxygen production quantum yield as well as high two-photon absorption cross-sections but with a very poor cellular internalization. We present herein new photosensitizers of the same porphyrin-triphenylamine hybrid series but bearing cationic charges which led to strongly enhanced water solubility and thus cellular penetration. In addition the new compounds have been found localized in mitochondria that are preferential target organelles for photodynamic therapy. Altogether the strongly improved properties of the new series combined with their specific mitochondrial localization lead to a significantly enhanced two-photon activated photodynamic therapy efficiency.

Mitochondria-targeted cationic porphyrin-triphenylamine hybrids for enhanced two-photon photodynamic therapy

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Année de publication : 2017

Ludivine Guyon, Marc Pirrotta, Katerina Duskova, Anton Granzhan, Marie-Paule Teulade-Fichou, David Monchaud (2017 Nov 15)

TWJ-Screen: an isothermal screening assay to assess ligand/DNA junction interactions in vitro

Nucleic Acids Research : 46 : e16 : DOI : 10.1093/nar/gkx1118 En savoir plus
Résumé

The quest for chemicals able to operate at selected genomic loci in a spatiotemporally controlled manner is desirable to create manageable DNA damages. Mounting evidence now shows that alternative DNA structures, including G-quadruplexes and branched DNA (or DNA junctions), might hamper proper progression of replication fork, thus triggering DNA damages and genomic instability. Therefore, small molecules that stabilize these DNA structures are currently scrutinized as a promising way to create genomic defects that cannot be dealt with properly by cancer cells. While much emphasis has been recently given to G-quadruplexes and related ligands, we report herein on three-way DNA junctions (TWJ) and related ligands. We first highlight the biological implications of TWJ and their strategic relevance as triggers for replicative stress. Then, we describe a new in vitro high-throughput screening assay, TWJ-Screen, which allows for identifying TWJ ligands with both high affinity and selectivity for TWJ over other DNA structures (duplexes and quadruplexes), in a convenient and unbiased manner as demonstrated by the screening of a library of 25 compounds from different chemical families. TWJ-Screen thus represents a reliable mean to uncover molecular tools able to foster replicative stress through an innovative approach, thus providing new strategic opportunities to combat cancers.

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Chiara Mauriello-Jimenez, Maxime Henry, Dina Aggad, Laurence Raehm, Xavier Cattoën, Michel Wong Chi Man, Clarence Charnay, Serkan Alpugan, Vefa Ahsen, Deniz Kutlu Tarakci, Philippe Maillard, Marie Maynadier, Marcel Garcia, Fabienne Dumoulin, Magali Gary-Bobo, Jean-Luc Coll, Véronique Josserand, Jean-Olivier Durand (2017 Oct 31)

Porphyrin- or phthalocyanine-bridged silsesquioxane nanoparticles for two-photon photodynamic therapy or photoacoustic imaging.

Nanoscale : 9 : 16622-16626 : DOI : 10.1039/c7nr04677d En savoir plus
Résumé

Porphyrin- or phthalocyanine-bridged silsesquioxane nanoparticles (BSPOR and BSPHT) were prepared. Their endocytosis in MCF-7 cancer cells was shown with two-photon excited fluorescence (TPEF) imaging. With two-photon excited photodynamic therapy (TPE-PDT), BSPOR was more phototoxic than BSPHT, which in contrast displayed a very high signal for photoacoustic imaging in mice.

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Michelle Newman, Rym Sfaxi, Abhijit Saha, David Monchaud, Marie-Paule Teulade-Fichou, Stéphan Vagner (2017 Oct 27)

The G-Quadruplex-Specific RNA Helicase DHX36 Regulates p53 Pre-mRNA 3′-End Processing Following UV-Induced DNA Damage.

Journal of Molecular Biology : 429 : 3121-3131 : DOI : 10.1016/j.jmb.2016.11.033 En savoir plus
Résumé

Pre-mRNA 3′-end processing, the process through which almost all eukaryotic mRNAs acquire a poly(A) tail is generally inhibited during the cellular DNA damage response leading to a profound impact on the level of protein expression since unprocessed transcripts at the 3′-end will be degraded or unable to be transported to the cytoplasm. However, a compensatory mechanism involving the binding of the hnRNP H/F family of RNA binding proteins to an RNA G-quadruplex (G4) structure located in the vicinity of a polyadenylation site has previously been described to allow the transcript encoding the p53 tumour suppressor protein to be properly processed during DNA damage and to provide the cells with a way to react to DNA damage. Here we report that the DEAH (Asp-Glu-Ala-His) box RNA helicase DHX36/RHAU/G4R1, which specifically binds to and resolves parallel-stranded G4, is necessary to maintain p53 pre-mRNA 3′-end processing following UV-induced DNA damage. DHX36 binds to the p53 RNA G4, while mutation of the G4 impairs the ability of DHX36 to maintain pre-mRNA 3′-end processing. Stabilization of the p53 RNA G4 with two different G4 ligands ((PNA)DOTASQ and PhenDC3), which is expected from previous studies to prevent DHX36 from binding and unwinding G4s, also impairs p53 pre-mRNA 3′-end processing following UV. Our work identifies DHX36 as a new actor in the compensatory mechanisms that are in place to ensure that the mRNAs encoding p53 are still processed following UV.

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Pabon-Martinez Y.V., Xu Y., Villa A., Lundin K.E., Geny S., Nguyen C.H., Pedersen E.B., Jorgensen P.T., Wengel J., Nilsson L., Smith C.I.E., Zain R. (2017 Sep 8)

LNA effects on DNA binding and conformation: from single strand to duplex and triplex structures

Scientific Reports : 7 : 11043 : DOI : 10.1038/s41598-017-09147-8 En savoir plus
Résumé

The anti-gene strategy is based on sequence-specific recognition of double-strand DNA by triplex forming (TFOs) or DNA strand invading oligonucleotides to modulate gene expression. To be efficient, the oligonucleotides (ONs) should target DNA selectively, with high affinity. Here we combined hybridization analysis and electrophoretic mobility shift assay with molecular dynamics (MD) simulations to better understand the underlying structural features of modified ONs in stabilizing duplex-and triplex structures. Particularly, we investigated the role played by the position and number of locked nucleic acid (LNA) substitutions in the ON when targeting a c-MYC or FXN (Frataxin) sequence. We found that LNA-containing single strand TFOs are conformationally pre-organized for major groove binding. Reduced content of LNA at consecutive positions at the 3′-end of a TFO destabilizes the triplex structure, whereas the presence of Twisted Intercalating Nucleic Acid (TINA) at the 3′-end of the TFO increases the rate and extent of triplex formation. A triplex-specific intercalating benzoquinoquinoxaline (BQQ) compound highly stabilizes LNA-containing triplex structures. Moreover, LNA-substitution in the duplex pyrimidine strand alters the double helix structure, affecting x-displacement, slide and twist favoring triplex formation through enhanced TFO major groove accommodation. Collectively, these findings should facilitate the design of potent anti-gene ONs.

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Pauline Gilson, Fernando Josa-Prado, Claire Beauvineau, Delphine Naud-Martin, Laetitia Vanwonterghem, Florence Mahuteau-Betzer, Alexis Moreno, Pierre Falson, Laurence Lafanechère, Véronique Frachet, Jean-Luc Coll, Jose Fernando Díaz, Amandine Hurbin, Benoit Busser (2017 Sep 2)

Identification of pyrrolopyrimidine derivative PP-13 as a novel microtubule-destabilizing agent with promising anticancer properties.

Scientific reports : 7 : 10209 : DOI : 10.1038/s41598-017-09491-9 En savoir plus
Résumé

Despite the emergence of targeted therapies and immunotherapy, chemotherapy remains the gold-standard for the treatment of most patients with solid malignancies. Spindle poisons that interfere with microtubule dynamics are commonly used in chemotherapy drug combinations. However, their troublesome side effects and the emergence of chemoresistance highlight the need for identifying alternative agents. We performed a high throughput cell-based screening and selected a pyrrolopyrimidine molecule (named PP-13). In the present study, we evaluated its anticancer properties in vitro and in vivo. We showed that PP-13 exerted cytotoxic effects on various cancer cells, including those resistant to current targeted therapies and chemotherapies. PP-13 induced a transient mitotic blockade by interfering with both mitotic spindle organization and microtubule dynamics and finally led to mitotic slippage, aneuploidy and direct apoptotic death. PP-13 was identified as a microtubule-targeting agent that binds directly to the colchicine site in β-tubulin. Interestingly, PP-13 overcame the multidrug-resistant cancer cell phenotype and significantly reduced tumour growth and metastatic invasiveness without any noticeable toxicity for the chicken embryo in vivo. Overall, PP-13 appears to be a novel synthetic microtubule inhibitor with interesting anticancer properties and could be further investigated as a potent alternative for the management of malignancies including chemoresistant ones.

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Foy J.P., Bazire L., Ortiz-Cuaran S., Deneuve S., Kielbassa J., Thomas E., Viari A., Puisieux A., Goudot P., Bertolus C., Foray N., Kirova Y., Verrelle P., Saintigny P. (2017 Sep 1)

A 13-gene expression-based radioresistance score highlights the heterogeneity in the response to radiation therapy across HPV-negative HNSCC molecular subtypes

BMC MEDICINE : 15 : 165 : DOI : 10.1186/s12916-017-0929-y En savoir plus
Résumé

Background: Radiotherapy for head and neck squamous cell carcinomas (HNSCC) is associated with a substantial morbidity and inconsistent efficacy. Human papillomavirus (HPV)-positive status is recognized as a marker of increased radiosensitivity. Our goal was to identify molecular markers associated with benefit to radiotherapy in patients with HPV-negative disease.

Methods: Gene expression profiles from public repositories were downloaded for data mining. Training sets included 421 HPV-negative HNSCC tumors from The Cancer Genome Atlas (TCGA) and 32 HNSCC cell lines with available radiosensitivity data (GSE79368). A radioresistance (RadR) score was computed using the single sample Gene Set Enrichment Analysis tool. The validation sets included two panels of cell lines (NCI-60 and GSE21644) and HPV-negative HNSCC tumor datasets, including 44 (GSE6631), 82 (GSE39366), and 179 (GSE65858) patients, respectively. We finally performed an integrated analysis of the RadR score with known recurrent genomic alterations in HNSCC, patterns of protein expression, biological hallmarks, and patterns of drug sensitivity using TCGA and the E-MTAB-3610 dataset (659 pancancer cell lines, 140 drugs).

Results: We identified 13 genes differentially expressed between tumor and normal head and neck mucosa that were associated with radioresistance in vitro and in patients. The 13-gene expression-based RadR score was associated with recurrence in patients treated with surgery and adjuvant radiotherapy but not with surgery alone. It was significantly different among different molecular subtypes of HPV-negative HNSCC and was significantly lower in the «  atypical » molecular subtype. An integrated analysis of RadR score with genomic alterations, protein expression, biological hallmarks and patterns of drug sensitivity showed a significant association with CCND1 amplification, fibronectin expression, seven hallmarks (including epithelial-to-mesenchymal transition and unfolded protein response), and increased sensitivity to elesclomol, an HSP90 inhibitor.

Conclusions: Our study highlights the clinical relevance of the molecular classification of HNSCC and the RadR score to refine radiation strategies in HPV-negative disease.

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Lista M.J., Martins R.P., Angrand G., Quillevere A., Daskalogianni C., Voisset C., Teulade-Fichou M.P., Fahraeus R., Blondel M. (2017 Sep 1)

A yeast model for the mechanism of the Epstein-Barr virus immune evasion identifies a new therapeutic target to interfere with the virus stealthiness

MICROBIAL CELL : 4 : 305-307 : DOI : 10.15698/mic2017.09.590 En savoir plus
Résumé

The oncogenic Epstein-Barr virus (EBV) evades the immune system but has an Achilles heel: its genome maintenance protein EBNA1. Indeed, EBNA1 is essential for viral genome replication and maintenance but also highly antigenic. Hence, EBV evolved a system in which the glycine-alanine repeat (GAr) of EBNA1 limits the translation of its own mRNA at a minimal level to ensure its essential function thereby, at the same time, minimizing immune recognition. Defining intervention points where to interfere with EBNA1 immune evasion is an important step to trigger an immune response against EBV-carrying cancers. Thanks to a yeast-based assay that recapitulates all the aspects of EBNA1 self-limitation of expression, a recent study by Lista et al. [Nature Communications (2017) 7, 435-444] has uncovered the role of the host cell nucleolin (NCL) in this process via a direct interaction of this protein with G-quadruplexes (G4) formed in GAr-encoding sequence of EBNA1 mRNA. In addition, the G4 ligand PhenDC3 prevents NCL binding on EBNA1 mRNA and reverses GAr-mediated repression of translation and antigen presentation. This shows that the NCL-EBNA1 mRNA interaction is a relevant therapeutic target to unveil EBV-carrying cancers to the immune system and that the yeast model can be successfully used for uncovering drugs and host factors that interfere with EBV stealthiness.

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Buchieri Maria V., Cimino Mena , Rebollo-Ramirez Sonia, Beauvineau Claire, Cascioferro Alessandro, Favre-Rochex Sandrine, Helynck Olivier, Naud-Martin Delphine, Larrouy-Maumus Gerald, Munier-Lehmann Hélène, Gicquel Brigitte (2017 Aug 28)

Nitazoxanide Analogs Require Nitroreduction for Antimicrobial Activity in Mycobacterium smegmatis

Journal of Medicinal Chemistry : 60 : 7425-7433 : DOI : 10.1021/acs.jmedchem.7b00726 En savoir plus
Résumé

In this study, we aimed to decipher the natural resistance mechanisms of mycobacteria against novel compounds isolated by whole-cell-based high-throughput screening (HTS). We identified active compounds using Mycobacterium aurum. Further analyses were performed to determine the resistance mechanism of M. smegmatis against one hit, 3-bromo-N-(5-nitrothiazol-2-yl)-4-propoxybenzamide (3), which turned out to be an analog of the drug nitazoxanide (1). We found that the repression of the gene nfnB coding for the nitroreductase NfnB was responsible for the natural resistance of M. smegmatis against 3. The overexpression of nfnB resulted in sensitivity of M. smegmatis to 3. This compound must be metabolized into hydroxylamine intermediate for exhibiting antibacterial activity. Thus, we describe, for the first time, the activity of a mycobacterial nitroreductase against 1 analogs, highlighting the differences in the metabolism of nitro compounds among mycobacterial species and emphasizing the potential of nitro drugs as antibacterials in various bacterial species.

Nitazoxanide-fig-abstract

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Aziz Jessy, Piguel Sandrine (2017 Aug 25)

An update on direct C–H bond functionalization of nitrogen-containing fused heterocycles

Synthesis : 49 : 4562-4585 : DOI : 10.1055/s-0036-1590859 En savoir plus
Résumé

This report highlights the recent advances in direct C–H bond functionalization of 5,5- and 6,5-fused heterocycles containing at least two nitrogen atoms. Besides C–C bond formation, C–N, C–S, C–P, and C–Si bonds can also be created via a metal-catalyzed process. Some examples, where a C–H functionalization approach was applied for the synthesis of drug candidates, will be presented as well.

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Claire Beauvineau, Corinne Guetta, Marie-Paule Teulade-Fichou, Florence Mahuteau-Betzer (2017 Aug 14)

PhenDV, a turn-off fluorescent quadruplex DNA probe for improving the sensitivity of drug screening assays

Organic & Biomolecular Chemistry : 15 : 7117-7121 : DOI : 10.1039/c7ob01705g En savoir plus
Résumé

We report a new turn-off fluorescent probe, PhenDV, for the identification of high affinity quadruplex (G4) DNA ligands. This push–pull fluorophore displays a high fluorescence quantum yield in water (ΦF = 0.21) and is a selective and strong quadruplex DNA binder. We describe its use as a fluorescent indicator for the G4 Fluorescent Intercalator Displacement (FID) assay as its fluorescence is strongly quenched when bound to G4 DNA and fully restored when displaced by ligand. This probe improves the sensitivity of the G4-FID assay, as the read out relies on increased fluorescence instead of quenching observed with classical on/off probes

PhenDV-Mahuteau-Betzer

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Jiyeon Choi, Mai Xu, Matthew M Makowski, Tongwu Zhang, Matthew H Law, Michael A Kovacs, Anton Granzhan, Wendy J Kim, Hemang Parikh, Michael Gartside, Jeffrey M Trent, Marie-Paule Teulade-Fichou, Mark M Iles, Julia A Newton-Bishop, D Timothy Bishop, Stuart MacGregor, Nicholas K Hayward, Michiel Vermeulen, Kevin M Brown (2017 Aug 1)

A common intronic variant of PARP1 confers melanoma risk and mediates melanocyte growth via regulation of MITF.

Nature genetics : 49 : 1326-1335 : DOI : 10.1038/ng.3927 En savoir plus
Résumé

Previous genome-wide association studies have identified a melanoma-associated locus at 1q42.1 that encompasses a ∼100-kb region spanning the PARP1 gene. Expression quantitative trait locus (eQTL) analysis in multiple cell types of the melanocytic lineage consistently demonstrated that the 1q42.1 melanoma risk allele (rs3219090[G]) is correlated with higher PARP1 levels. In silico fine-mapping and functional validation identified a common intronic indel, rs144361550 (-/GGGCCC; r(2) = 0.947 with rs3219090), as displaying allele-specific transcriptional activity. A proteomic screen identified RECQL as binding to rs144361550 in an allele-preferential manner. In human primary melanocytes, PARP1 promoted cell proliferation and rescued BRAF(V600E)-induced senescence phenotypes in a PARylation-independent manner. PARP1 also transformed TERT-immortalized melanocytes expressing BRAF(V600E). PARP1-mediated senescence rescue was accompanied by transcriptional activation of the melanocyte-lineage survival oncogene MITF, highlighting a new role for PARP1 in melanomagenesis.

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Verga D., Hamon F., Nicoleau C., Guetta C., Wu T.-D., Guerquin-Kern J.-L., Marco S. & Teulade-Fichou M.-P. (2017 Jul 27)

Chemical Imaging by NanoSIMS Provides High- Resolution Localization of the G-Quadruplex Interactive Drug (Br)-PhenDC3 on Human Chromosomes

Journal of Molecular Biology and Molecular Imaging : 4 : 1029 En savoir plus
Résumé

Determining the distribution of biologically active compounds within cells is a major issue to understand their mechanism of action and to optimize their properties. Over the past decade DNA secondary structures called G-quadruplexes (G4) have been identified as key modulators of genomic functions. This very active research field has led to the development of G4-targeted molecular probes that are used to track quadruplex forming domains in cells, which is achieved, in most cases, by conventional fluorescence microcopy. However, the intrinsic low resolution of fluorescence microcopy as well as the necessity to tag the drugs with fluorophores represent strong limitations. Here we present the use of secondary ion mass spectroscopy imaging (nanoSIMS) for mapping within metaphase human chromosomes the distribution of a bromobisquinolinium phenanthroline derivative (Br- PhenDC3) used as G-quadruplex probe. In addition a statistical approach to increase the accuracy and the spatial resolution of the nanoSIMS imaging was implemented as a plug in for the image analysis software ImageJ. The results demonstrate the presence of Br-PhenDC3both at terminal and interstitial regions of chromosomes and constitute a demonstration of the effectiveness of nanoSIMS imaging as an alternative method for accurate genome-wide mapping of DNA interactive drugs.

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