Diversité et plasticité des tumeurs de l’enfant

Publications de l’équipe

Année de publication : 2019

Angela Bellini, Nadia Bessoltane-Bentahar, Jaydutt Bhalshankar, Nathalie Clement, Virginie Raynal, Sylvain Baulande, Virginie Bernard, Adrien Danzon, Mathieu Chicard, Léo Colmet-Daage, Gaelle Pierron, Laura Le Roux, Julien Masliah Planchon, Valérie Combaret, Eve Lapouble, Nadège Corradini, Estelle Thebaud, Marion Gambart, Dominique Valteau-Couanet, Jean Michon, Caroline Louis-Brennetot, Isabelle Janoueix-Lerosey, Anne-Sophie Defachelles, Franck Bourdeaut, Olivier Delattre, Gudrun Schleiermacher (2019 Apr 25)

Study of chromatin remodeling genes implicates SMARCA4 as a putative player in oncogenesis in neuroblastoma.

International journal of cancer : DOI : 10.1002/ijc.32361 En savoir plus
Résumé

In neuroblastoma (NB), genetic alterations in chromatin remodeling (CRGs) and epigenetic modifier genes (EMGs) have been described. We sought to determine their frequency and clinical impact. Whole exome (WES)/whole genome sequencing (WGS) data and targeted sequencing (TSCA®) of exonic regions of 33 CRGs/EMGs were analyzed in tumor samples from 283 NB patients, with constitutional material available for 55 patients. The frequency of CRG/EMG variations in NB cases was then compared to the Genome Aggregation Database (gnomAD). The sequencing revealed SNVs/small InDels or focal CNAs of CRGs/EMGs in 20% (56/283) of all cases, occurring at a somatic level in 4 (7.2%), at a germline level in 12 (22%) cases, whereas for the remaining cases, only tumor material could be analyzed. The most frequently altered genes were ATRX (5%), SMARCA4 (2.5%), MLL3 (2.5%) and ARID1B (2.5%). Double events (SNVs/small InDels/CNAs associated with LOH) were observed in SMARCA4 (n=3), ATRX (n=1) and PBRM1 (n=1). Among the 60 variations, 24 (8.4%) targeted domains of functional importance for chromatin remodeling or highly conserved domains but of unknown function. Variations in SMARCA4 and ATRX occurred more frequently in the NB as compared to the gnomAD control cohort (OR=4.49, 95%CI:1.63-9.97, P=0.038; OR 3.44, 95%CI:1.46-6.91, P=0.043, respectively). Cases with CRG/EMG variations showed a poorer overall survival compared to cases without variations. Genetic variations of CRGs/EMGs with likely functional impact were observed in 8.4% (24/283) of NB. Our case-control approach suggests a role of SMARCA4 as a player of NB oncogenesis. This article is protected by copyright. All rights reserved.

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Année de publication : 2018

Mitchell J Machiela, Thomas G P Grünewald, Didier Surdez, Stephanie Reynaud, Olivier Mirabeau, Eric Karlins, Rebeca Alba Rubio, Sakina Zaidi, Sandrine Grossetete-Lalami, Stelly Ballet, Eve Lapouble, Valérie Laurence, Jean Michon, Gaelle Pierron, Heinrich Kovar, Nathalie Gaspar, Udo Kontny, Anna González-Neira, Piero Picci, Javier Alonso, Ana Patino-Garcia, Nadège Corradini, Perrine Marec Bérard, Neal D Freedman, Nathaniel Rothman, Casey L Dagnall, Laurie Burdett, Kristine Jones, Michelle Manning, Kathleen Wyatt, Weiyin Zhou, Meredith Yeager, David G Cox, Robert N Hoover, Javed Khan, Gregory T Armstrong, Wendy M Leisenring, Smita Bhatia, Leslie L Robison, Andreas E Kulozik, Jennifer Kriebel, Thomas Meitinger, Markus Metzler, Wolfgang Hartmann, Konstantin Strauch, Thomas Kirchner, Uta Dirksen, Lindsay M Morton, Lisa Mirabello, Margaret A Tucker, Franck Tirode, Stephen J Chanock, Olivier Delattre (2018 Aug 11)

Genome-wide association study identifies multiple new loci associated with Ewing sarcoma susceptibility.

Nature communications : 3184 : DOI : 10.1038/s41467-018-05537-2 En savoir plus
Résumé

Ewing sarcoma (EWS) is a pediatric cancer characterized by the EWSR1-FLI1 fusion. We performed a genome-wide association study of 733 EWS cases and 1346 unaffected individuals of European ancestry. Our study replicates previously reported susceptibility loci at 1p36.22, 10q21.3 and 15q15.1, and identifies new loci at 6p25.1, 20p11.22 and 20p11.23. Effect estimates exhibit odds ratios in excess of 1.7, which is high for cancer GWAS, and striking in light of the rarity of EWS cases in familial cancer syndromes. Expression quantitative trait locus (eQTL) analyses identify candidate genes at 6p25.1 (RREB1) and 20p11.23 (KIZ). The 20p11.22 locus is near NKX2-2, a highly overexpressed gene in EWS. Interestingly, most loci reside near GGAA repeat sequences and may disrupt binding of the EWSR1-FLI1 fusion protein. The high locus to case discovery ratio from 733 EWS cases suggests a genetic architecture in which moderate risk SNPs constitute a significant fraction of risk.

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Kathleen I Pishas, Christina D Drenberg, Cenny Taslim, Emily R Theisen, Kirsten M Johnson, Ranajeet S Saund, Ioana L Pop, Brian D Crompton, Elizabeth R Lawlor, Franck Tirode, Jaume Mora, Olivier Delattre, Mary C Beckerle, David F Callen, Sunil Sharma, Stephen L Lessnick (2018 Jul 13)

Therapeutic Targeting of KDM1A/LSD1 in Ewing Sarcoma with SP-2509 Engages the Endoplasmic Reticulum Stress Response.

Molecular cancer therapeutics : 1902-1916 : DOI : 10.1158/1535-7163.MCT-18-0373 En savoir plus
Résumé

Multi-agent chemotherapeutic regimes remain the cornerstone treatment for Ewing sarcoma, the second most common bone malignancy diagnosed in pediatric and young adolescent populations. We have reached a therapeutic ceiling with conventional cytotoxic agents, highlighting the need to adopt novel approaches that specifically target the drivers of Ewing sarcoma oncogenesis. As KDM1A/ysine-pecific emethylase 1 (LSD1) is highly expressed in Ewing sarcoma cell lines and tumors, with elevated expression levels associated with worse overall survival ( = 0.033), this study has examined biomarkers of sensitivity and mechanisms of cytotoxicity to targeted inhibition using SP-2509 (reversible inhibitor). We report, that innate resistance to SP-2509 was not observed in our Ewing sarcoma cell line cohort ( = 17; IC range, 81 -1,593 nmol/L), in contrast resistance to the next-generation irreversible inhibitor GSK-LSD1 was observed across multiple cell lines (IC > 300 μmol/L). Although status and basal KDM1A mRNA and protein levels did not correlate with SP-2509 response, induction of KDM1B following SP-2509 treatment was strongly associated with SP-2509 hypersensitivity. We show that the transcriptional profile driven by SP-2509 strongly mirrors genetic depletion. Mechanistically, RNA-seq analysis revealed that SP-2509 imparts robust apoptosis through engagement of the endoplasmic reticulum stress pathway. In addition, were specifically induced/repressed, respectively following SP-2509 treatment only in our hypersensitive cell lines. Together, our findings provide key insights into the mechanisms of SP-2509 cytotoxicity as well as biomarkers that can be used to predict inhibitor sensitivity in Ewing sarcoma. .

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Thomas G P Grünewald, Florencia Cidre-Aranaz, Didier Surdez, Eleni M Tomazou, Enrique de Álava, Heinrich Kovar, Poul H Sorensen, Olivier Delattre, Uta Dirksen (2018 Jul 7)

Ewing sarcoma.

Nature reviews. Disease primers : 5 : DOI : 10.1038/s41572-018-0003-x En savoir plus
Résumé

Ewing sarcoma is the second most frequent bone tumour of childhood and adolescence that can also arise in soft tissue. Ewing sarcoma is a highly aggressive cancer, with a survival of 70-80% for patients with standard-risk and localized disease and ~30% for those with metastatic disease. Treatment comprises local surgery, radiotherapy and polychemotherapy, which are associated with acute and chronic adverse effects that may compromise quality of life in survivors. Histologically, Ewing sarcomas are composed of small round cells expressing high levels of CD99. Genetically, they are characterized by balanced chromosomal translocations in which a member of the FET gene family is fused with an ETS transcription factor, with the most common fusion being EWSR1-FLI1 (85% of cases). Ewing sarcoma breakpoint region 1 protein (EWSR1)-Friend leukaemia integration 1 transcription factor (FLI1) is a tumour-specific chimeric transcription factor (EWSR1-FLI1) with neomorphic effects that massively rewires the transcriptome. Additionally, EWSR1-FLI1 reprogrammes the epigenome by inducing de novo enhancers at GGAA microsatellites and by altering the state of gene regulatory elements, creating a unique epigenetic signature. Additional mutations at diagnosis are rare and mainly involve STAG2, TP53 and CDKN2A deletions. Emerging studies on the molecular mechanisms of Ewing sarcoma hold promise for improvements in early detection, disease monitoring, lower treatment-related toxicity, overall survival and quality of life.

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Irene Jiménez, Mathieu Chicard, Léo Colmet-Daage, Nathalie Clément, Adrien Danzon, Eve Lapouble, Gaelle Pierron, Mylène Bohec, Sylvain Baulande, Dominique Berrebi, Paul Fréneaux, Aurore Coulomb, Louise Galmiche-Rolland, Sabine Sarnacki, Georges Audry, Pascale Philippe-Chomette, Hervé J Brisse, François Doz, Jean Michon, Olivier Delattre, Gudrun Schleiermacher (2018 Jun 21)

Circulating tumor DNA analysis enables molecular characterization of pediatric renal tumors at diagnosis.

International journal of cancer : DOI : 10.1002/ijc.31620 En savoir plus
Résumé

Circulating tumor DNA (ctDNA) is a powerful tool for the molecular characterization of cancer. The most frequent pediatric kidney tumors (KT) are Wilms’ tumors (WT), but other diagnoses may occur. According to the SIOP strategy, in most countries pediatric KT have a presumptive diagnosis of WT if they are clinically and radiologically compatible. The histologic confirmation is established after post-chemotherapy nephrectomy. Thus, there is a risk for a small fraction of patients to receive neoadjuvant chemotherapy that is not adapted to the disease. The aim of this work is to perform molecular diagnosis of pediatric KT by tumor genetic characterization based on the analysis of ctDNA. We analyzed ctDNA extracted from plasma samples of 18 pediatric patients with KT by whole-exome sequencing and compared the results to their matched tumor and germline DNA. Copy number alterations (CNAs) and single nucleotide variations (SNVs) were analyzed. We were able to detect tumor cell specific genetic alterations-CNAs, SNVs or both-in ctDNA in all patients except in one (for whom the plasma sample was obtained long after nephrectomy). These results open the door to new applications for the study of ctDNA with regards to the molecular diagnosis of KT, with a possibility of its usefulness for adapting the treatment early after diagnosis, but also for disease monitoring and follow up.

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Genevieve C Kendall, Sarah Watson, Lin Xu, Collette A LaVigne, Whitney Murchison, Dinesh Rakheja, Stephen X Skapek, Franck Tirode, Olivier Delattre, James F Amatruda (2018 Jun 6)

transgenic zebrafish models identify as a mediator of rhabdomyosarcoma tumorigenesis.

eLife : DOI : 10.7554/eLife.33800 En savoir plus
Résumé

Alveolar rhabdomyosarcoma is a pediatric soft-tissue sarcoma caused by fusion oncogenes and is characterized by impaired skeletal muscle development. We developed human -driven zebrafish models of tumorigenesis and found that exhibits discrete cell lineage susceptibility and transformation. Tumors developed by 1.6-19 months and were primitive neuroectodermal tumors or rhabdomyosarcoma. We applied this transgenic zebrafish model to study how leverages early developmental pathways for oncogenesis and found that is a unique target. Ectopic expression of the human ortholog, , inhibits myogenesis in zebrafish and mammalian cells, recapitulating the arrested muscle development characteristic of rhabdomyosarcoma. In patients, is overexpressed in fusion-positive versus fusion-negative tumors. Finally, overexpression is associated with reduced survival in patients in the context of the fusion. Our novel zebrafish rhabdomyosarcoma model identifies a new target, /, that contributes to impaired myogenic differentiation and has prognostic significance in human disease.

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Deveau, P., Colmet Daage, L., Oldridge, D., Bernard, V., Bellini, A., Chicard, M., Clement, N., Lapouble, E., Combaret, V., Boland, A., Meyer, V., Deleuze, J.-F., Janoueix-Lerosey, I., Barillot, E., Delattre, O., Maris, J.M., Schleiermacher, G., and Boeva, V (2018 Jun 1)

QuantumClone: clonal assessment of functional mutations in cancer based on a genotype-aware method for clonal reconstruction

Bioinformatics : 34 : 1808,1816 : DOI : 10.1093/bioinformatics/bty016. En savoir plus
Résumé

Motivation:

In cancer, clonalevolution is assessed basedon information coming from single nucleotide variants and copy number alterations. Nonetheless, existing methods often fail to accurately combine information from both sources to truthfully reconstruct clonalpopulations in a given tumor sample or in a set of tumor samples coming from the same patient. Moreover, previously published methods detect clones from a single set of variants. As a result, compromises have to be done between stringent variant filtering [reducing dispersion in variant allele frequency estimates (VAFs)] and using all biologically relevant variants.

Results:

We present a framework for defining cancerclones using most reliable variants of high depth of coverage and assigning functionalmutationsto the detected clones. The key element of our framework is QuantumClone, a methodfor variant clustering into clones basedon VAFs, genotypes of corresponding regions and information about tumor purity. We validated QuantumCloneand our framework on simulated data. We then applied our framework to whole genome sequencing data for 19 neuroblastoma trios each including constitutional, diagnosis and relapse samples. We confirmed an enrichment of damaging variants within such pathways as MAPK (mitogen-activated protein kinases), neuritogenesis, epithelial-mesenchymal transition, cell survival and DNA repair. Most pathways had more damaging variants in the expanding clones compared to shrinking ones, which can be explained by the increased total number of variants between these two populations. Functionalmutational rate varied for ancestral clones and clones shrinking or expanding upon treatment, suggesting changes in clone selection mechanisms at different time points of tumor evolution.

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Franck Bourdeaut, Olivier Delattre (2018 May 14)

Genetic predisposition to medulloblastomas: just follow the tumour genome.

The Lancet. Oncology : 722-723 : DOI : S1470-2045(18)30289-4 En savoir plus
Résumé

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Janoueix-Lerosey, I., Lopez-Delisle, L., Delattre, O., and Rohrer, H (2018 May 2)

The ALK receptor in sympathetic neuron development and neuroblastoma

Cell and Tissue Research : 325, 337 : DOI : https://doi.org/10.1007/s00441-017-2784-8 En savoir plus
Résumé

The ALKgene encodes a tyrosine kinase receptorcharacterized by an expression pattern mainly restricted to the developing central and peripheral nervous systems. In 2008, the discovery of ALKactivating mutations in neuroblastoma, a tumor of the sympatheticnervous system, represented a breakthrough in the understanding of the pathogenesis of this pediatric cancer and established mutated ALKas a tractable therapeutic target for precision medicine. Subsequent studies addressed the identity of ALKligands, as well as its physiological function in the sympathoadrenal lineage, its role in neuroblastomadevelopmentand the signaling pathways triggered by mutated ALK. This review focuses on these different aspects of the ALKbiology and summarizes the various therapeutic strategies relying on ALKinhibition in neuroblastoma, either as monotherapies or combinatory treatments.

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Julien Masliah-Planchon, Dominique Lévy, Delphine Héron, Fabienne Giuliano, Catherine Badens, Paul Fréneaux, Louise Galmiche, Jean-Marc Guinebretierre, Cécile Cellier, Joshua J Waterfall, Khadija Aït-Raïs, Gaëlle Pierron, Christophe Glorion, Isabelle Desguerre, Christine Soler, Anne Deville, Olivier Delattre, Jean Michon, Franck Bourdeaut (2018 May 1)

Does ATRX germline variation predispose to osteosarcoma? Three additional cases of osteosarcoma in two ATR-X syndrome patients.

European journal of human genetics : EJHG : 1217-1221 : DOI : 10.1038/s41431-018-0147-x En savoir plus
Résumé

Osteosarcoma is the most common malignant bone tumor in adolescents and young adults. Most osteosarcomas are sporadic but the risk of osteosarcoma is also increased by germline variants in TP53, RB1 and RECQL4 genes. ATRX germline variations are responsible for the rare genetic disorder X-linked alpha-thalassemia mental retardation (ATR-X) syndrome characterized by severe developmental delay and alpha-thalassemia but no obvious increased risk of cancer. Here we report two children with ATR-X syndrome who developed osteosarcoma. Notably, one of the children developed two osteosarcomas separated by 10 years. Those two cases raise the possibility that ATRX germline variant could be associated with an increased risk of osteosarcoma.

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Watson, S., Perrin, V., Guillemot, D., Reynaud, S., Coindre, J.-M., Karanian, M., Guinebretière, J.-M., Freneaux, P., Le Loarer, F., Bouvet, M., Galmiche-Rolland, L., Larousserie, F., Longchampt, E., Ranchere-Vince, D., Pierron, G., Delattre, O., and Tirode, F (2018 Mar 30)

Transcriptomic definition of molecular subgroups of small round cell sarcomas

The Journal of Pathology : 245 : 29, 40 : DOI : doi.org/10.1002/path.5053 En savoir plus
Résumé

Sarcoma represents a highly heterogeneous group of tumours. We report here the first unbiased and systematic search for gene fusions combined with unsupervised expression analysis of a series of 184 small round cell sarcomas. Fusion genes were detected in 59% of samples, with half of them being observed recurrently. We identified biologically homogeneous groups of tumours such as the CIC‐fused (to DUX4, FOXO4or NUTM1) and BCOR‐rearranged (BCORCCNB3, BCORMAML3, ZC3H7B–BCOR, and BCORinternal duplication) tumour groups. VGLL2‐fused tumours represented a more biologically and pathologically heterogeneous group. This study also refined the characteristics of some entities such as EWSR1–PATZ1spindle cell sarcoma or FUS–NFATC2bone tumours that are different from EWSR1–NFATC2tumours and transcriptionally resemble CIC‐fused tumour entities. We also describe a completely novel group of epithelioid and spindle‐cell rhabdomyosarcomas characterized by EWSR1–or FUS–TFCP2fusions. Finally, expression data identified some potentially new therapeutic targets or pathways. Copyright © 2018 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.

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Gregorio J Petrirena, Julien Masliah-Planchon, Quentin Sala, Bertrand Pourroy, Didier Frappaz, Emeline Tabouret, Thomas Graillon, Jean-Claude Gentet, Olivier Delattre, Olivier Chinot, Laetitia Padovani (2018 Mar 9)

Recurrent extraneural sonic hedgehog medulloblastoma exhibiting sustained response to vismodegib and temozolomide monotherapies and inter-metastatic molecular heterogeneity at progression.

Oncotarget : 10175-10183 : DOI : 10.18632/oncotarget.23699 En savoir plus
Résumé

Response to targeting and non-targeting agents is variable and molecular information remains poorly described in patients with recurrent sonic-hedgehog-driven medulloblastoma (SHH-MB).

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Veronica Ferrucci, Pasqualino de Antonellis, Francesco Paolo Pennino, Fatemeh Asadzadeh, Antonella Virgilio, Donatella Montanaro, Aldo Galeone, Iolanda Boffa, Ida Pisano, Iolanda Scognamiglio, Luigi Navas, Donatella Diana, Emilia Pedone, Sara Gargiulo, Matteo Gramanzini, Arturo Brunetti, Laura Danielson, Marianeve Carotenuto, Lucia Liguori, Antonio Verrico, Lucia Quaglietta, Maria Elena Errico, Valentina Del Monaco, Valeria D'Argenio, Felice Tirone, Angela Mastronuzzi, Vittoria Donofrio, Felice Giangaspero, Daniel Picard, Marc Remke, Livia Garzia, Craig Daniels, Olivier Delattre, Fredrik J Swartling, William A Weiss, Francesco Salvatore, Roberto Fattorusso, Louis Chesler, Michael D Taylor, Giuseppe Cinalli, Massimo Zollo (2018 Mar 1)

Metastatic group 3 medulloblastoma is driven by PRUNE1 targeting NME1-TGF-β-OTX2-SNAIL via PTEN inhibition.

Brain : a journal of neurology : 1300-1319 : DOI : 10.1093/brain/awy039 En savoir plus
Résumé

Genetic modifications during development of paediatric groups 3 and 4 medulloblastoma are responsible for their highly metastatic properties and poor patient survival rates. PRUNE1 is highly expressed in metastatic medulloblastoma group 3, which is characterized by TGF-β signalling activation, c-MYC amplification, and OTX2 expression. We describe the process of activation of the PRUNE1 signalling pathway that includes its binding to NME1, TGF-β activation, OTX2 upregulation, SNAIL (SNAI1) upregulation, and PTEN inhibition. The newly identified small molecule pyrimido-pyrimidine derivative AA7.1 enhances PRUNE1 degradation, inhibits this activation network, and augments PTEN expression. Both AA7.1 and a competitive permeable peptide that impairs PRUNE1/NME1 complex formation, impair tumour growth and metastatic dissemination in orthotopic xenograft models with a metastatic medulloblastoma group 3 cell line (D425-Med cells). Using whole exome sequencing technology in metastatic medulloblastoma primary tumour cells, we also define 23 common ‘non-synonymous homozygous’ deleterious gene variants as part of the protein molecular network of relevance for metastatic processes. This PRUNE1/TGF-β/OTX2/PTEN axis, together with the medulloblastoma-driver mutations, is of relevance for future rational and targeted therapies for metastatic medulloblastoma group 3.10.1093/brain/awy039_video1awy039media15742053534001.

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Lopez-Delisle, L., Pierre-Eugène, C., Louis-Brennetot, C., Surdez, D., Raynal, V., Baulande, S., Boeva, V., Grossetête-Lalami, S., Combaret, V., Peuchmaur, M., Delattre, O., and Janoueix-Lerosey, I (2018 Jan 11)

Activated ALK signals through the ERK-ETV5-RET pathway to drive neuroblastoma oncogenesis

Oncogene : 37 : 1417, 1429 : DOI : doi.org/10.1038/s41388-017-0039-5 En savoir plus
Résumé

Activating mutations of the ALKreceptor occur in a subset of neuroblastomatumors. We previously demonstrated that Alkmutations cooperate with MYCN overexpression to induce neuroblastomain mice and identified Ret as being strongly upregulated in MYCN/Alkmuttumors. By a genetic approach in vivo, we now document an oncogenic cooperation between activatedRet and MYCN overexpression in neuroblastomaformation. We show that MYCN/RetM919Ttumors exhibit histological features and expression profiles close to MYCN/Alkmuttumors. We show that RET transcript levels decrease precedes RET protein levels decrease upon ALKinhibition in neuroblastomacell lines. Etv5 was identified as a candidate transcription factor regulating Ret expression from murine MYCN/Alkmuttumor transcriptomic data. We demonstrate that ETV5 is regulated both at the protein and mRNA levels upon ALKactivation or inhibition in neuroblastomacell lines and that this regulation precedes RET modulation. We document that ALKactivation induces ETV5 protein upregulation through stabilization in a MEK/ERK-dependent manner. We show that RNAi-mediated inhibition of ETV5 decreases RET expression. Reporter assays indicate that ETV5 is able todriveRET gene transcription. ChIP-seq analysis confirmed ETV5 binding on the RET promoter and identified an enhancer upstream of the promoter. Finally, we demonstrate that combining RET and ALKinhibitors reduces tumor growth more efficiently than each single agent in MYCN and AlkF1178L-driven murine neuroblastoma. Altogether, these results define the ERK-ETV5-RETpathwayas a critical axis driving neuroblastomaoncogenesisdownstream of activatedALK.

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Année de publication : 2017

Mathieu Chicard, Leo Colmet-Daage, Nathalie Clement, Adrien Danzon, Mylène Bohec, Virginie Bernard, Sylvain Baulande, Angela Bellini, Paul Deveau, Gaëlle Pierron, Eve Lapouble, Isabelle Janoueix-Lerosey, Michel Peuchmaur, Nadège Corradini, Anne Sophie Defachelles, Dominique Valteau-Couanet, Jean Michon, Valérie Combaret, Olivier Delattre, Gudrun Schleiermacher (2017 Dec 2)

Whole-Exome Sequencing of Cell-Free DNA Reveals Temporo-spatial Heterogeneity and Identifies Treatment-Resistant Clones in Neuroblastoma.

Clinical cancer research : an official journal of the American Association for Cancer Research : 939-949 : DOI : 10.1158/1078-0432.CCR-17-1586 En savoir plus
Résumé

Neuroblastoma displays important clinical and genetic heterogeneity, with emergence of new mutations at tumor progression. To study clonal evolution during treatment and follow-up, an innovative method based on circulating cell-free DNA (cfDNA) analysis by whole-exome sequencing (WES) paired with target sequencing was realized in sequential liquid biopsy samples of 19 neuroblastoma patients. WES of the primary tumor and cfDNA at diagnosis showed overlap of single-nucleotide variants (SNV) and copy number alterations, with 41% and 93% of all detected alterations common to the primary neuroblastoma and cfDNA. CfDNA WES at a second time point indicated a mean of 22 new SNVs for patients with progressive disease. Relapse-specific alterations included genes of the MAPK pathway and targeted the protein kinase A signaling pathway. Deep coverage target sequencing of intermediate time points during treatment and follow-up identified distinct subclones. For 17 seemingly relapse-specific SNVs detected by cfDNA WES at relapse but not tumor or cfDNA WES at diagnosis, deep coverage target sequencing detected these alterations in minor subclones, with relapse-emerging SNVs targeting genes of neuritogenesis and cell cycle. Furthermore a persisting, resistant clone with concomitant disappearance of other clones was identified by a mutation in the ubiquitin protein ligase Modelization of mutated allele fractions in cfDNA indicated distinct patterns of clonal evolution, with either a minor, treatment-resistant clone expanding to a major clone at relapse, or minor clones collaborating toward tumor progression. Identification of treatment-resistant clones will enable development of more efficient treatment strategies. .

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