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A Metabolic Function of FGFR3-TACC3 Gene Fusions In A metabolic function of FGFR3-TACC3 gene fusions in cancer Véronique Frattini, Stefano Pagnotta, Dr Tala, Jerry Fan, Marco Russo, Sang Lee, Luciano Garofano, Jing Zhang, Peiguo Shi, Genevieve Lewis, et al. To cite this version: Véronique Frattini, Stefano Pagnotta, Dr Tala, Jerry Fan, Marco Russo, et al.. A metabolic function of FGFR3-TACC3 gene fusions in cancer. Nature, Nature Publishing Group, 2018, 553 (7687), pp.222- 227. 10.1038/nature25171. hal-01975294 HAL Id: hal-01975294 https://hal.sorbonne-universite.fr/hal-01975294 Submitted on 29 Aug 2019 HAL is a multi-disciplinary open access L’archive ouverte pluridisciplinaire HAL, est archive for the deposit and dissemination of sci- destinée au dépôt et à la diffusion de documents entific research documents, whether they are pub- scientifiques de niveau recherche, publiés ou non, lished or not. The documents may come from émanant des établissements d’enseignement et de teaching and research institutions in France or recherche français ou étrangers, des laboratoires abroad, or from public or private research centers. publics ou privés. A metabolic function of FGFR3-TACC3 gene fusions in cancer Véronique Frattini1*, Stefano M. Pagnotta1,2*, Tala1, Jerry J. Fan3,4, Marco V. Russo1, Sang Bae Lee1, Luciano Garofano1,2,5, Jing Zhang1, Peiguo Shi1, Genevieve Lewis1, Heloise Sanson1, Vanessa Frederick1, Angelica M. Castano1, Luigi Cerulo2,5, Delphine C. M. Rolland6, Raghvendra Mall7, Karima Mokhtari8,9,10, Kojo S. J. Elenitoba-Johnson6, Marc Sanson8,10,11, Xi Huang3,4, Michele Ceccarelli2,5, Anna Lasorella1,12,13§ & Antonio Iavarone1,12,14§ Chromosomal translocations that generate in-frame oncogenic Data Fig. 1c and Supplementary Table 1). We confirmed the expression gene fusions are notable examples of the success of targeted cancer changes of mitochondrial genes by quantitative PCR with reverse tran- therapies1–3. We have previously described gene fusions of FGFR3- scription (RT–qPCR) (Extended Data Fig. 1d). TACC3 (F3–T3) in 3% of human glioblastoma cases4. Subsequent Compared to human astrocytes expressing the empty vector or studies have reported similar frequencies of F3–T3 in many other F3–T3(K508M), F3–T3 human astrocytes exhibited increased levels cancers, indicating that F3–T3 is a commonly occuring fusion of mitochondrial DNA, mitochondrial mass (MitoTracker Red) and across all tumour types5,6. F3–T3 fusions are potent oncogenes produced higher levels of ATP (Fig. 1c, d and Extended Data Fig. 1e). that confer sensitivity to FGFR inhibitors, but the downstream F3–T3 increased respiratory complex proteins (SDHB, UQCRC1 oncogenic signalling pathways remain unknown2,4–6. Here we and ATP5A1) and the mitochondrial membrane transporter VDAC1 show that human tumours with F3–T3 fusions cluster within (Extended Data Fig. 1f). We also found higher levels of VDAC1 and transcriptional subgroups that are characterized by the activation of NDUFS4 in tumours generated from mouse glioma stem cells (mGSCs) mitochondrial functions. F3–T3 activates oxidative phosphorylation expressing human F3–T3 and small hairpin RNA (shRNA) against and mitochondrial biogenesis and induces sensitivity to inhibitors of Trp53 (shTrp53) (hereafter F3–T3;shTrp53) than in tumours formed oxidative metabolism. Phosphorylation of the phosphopeptide PIN4 by mGSCs expressing oncogenic HRAS(12V) and shTrp53 (hereafter is an intermediate step in the signalling pathway of the activation HRAS(12V);shTrp53)4,7 (Extended Data Fig. 1g). Introduction of F3–T3 of mitochondrial metabolism. The F3–T3–PIN4 axis triggers the in human astrocytes, RPE and U251 cells increased the basal and biogenesis of peroxisomes and the synthesis of new proteins. The maximal oxygen consumption rate (OCR) of these cells compared to anabolic response converges on the PGC1α coactivator through the cells transduced with F3–T3(K508M) or empty vector and this effect production of intracellular reactive oxygen species, which enables was reversed by FGFR-TK inhibition with AZD4547 in cells expressing mitochondrial respiration and tumour growth. These data illustrate exogenous F3–T3 and human glioblastoma (GBM)-derived GSC1123 the oncogenic circuit engaged by F3–T3 and show that F3–T3- cells with endogenous F3–T34 (Fig. 1e and Extended Data Fig. 2a–d). positive tumours rely on mitochondrial respiration, highlighting F3–T3 elicited only a mild increase in the extracellular acidification this pathway as a therapeutic opportunity for the treatment of rate (ECAR), leading to an increase in the OCR:ECAR ratio (Extended tumours with F3–T3 fusions. We also provide insights into the Data Fig. 2e, f). After treatment with the inhibitor of ATP synthase genetic alterations that initiate the chain of metabolic responses oligomycin, F3–T3 human astrocytes displayed reduced ATP levels that drive mitochondrial metabolism in cancer. and cell growth (by more than 70%) but were resistant to the substi- To investigate the transcriptional changes elicited by F3–T3, we tution of glucose with galactose in the culture medium, a condition expressed F3–T3 in immortalized human astrocytes and compared that imposes oxidative metabolism and markedly affected cell growth gene expression profiles of cells treated with a specific inhibitor against of human astrocytes treated with vector (Extended Data Fig. 2g, h). A FGFR tyrosine kinase (TK, PD173074) or vehicle. Human astrocytes 72-h treatment with the mitochondrial inhibitors metformin, mena- expressing F3–T3 were also compared to human astrocytes that dione or tigecycline impaired growth of GSC1123 cells but was inef- expressed kinase-dead F3–T3 (F3–T3(K508M)) or were transduced fective in GSC308 F3–T3-negative gliomaspheres4 (Fig. 1f). Similarly, with an empty vector (Extended Data Fig. 1a). Hierarchical clustering mitochondrial inhibitors reduced the viability of F3–T3;shTrp53 based on genes that were differentially expressed between F3–T3 mGSCs but did not affect HRAS(12V);shTrp53 mGSCs (Fig. 1g and human astrocytes and PD173074-treated F3–T3 cells showed that Extended Data Fig. 2i–k). However, tigecycline decreased COX1 and F3–T3 human astrocytes differed from the other three groups (Fig. 1a COX2, two respiratory complex subunits translated by mitochondrial and Extended Data Fig. 1b). Analysis of a Gene Ontology enrichment ribosomes8, and mitochondrial inhibitors reduced ATP production, map showed that, in addition to the expected enrichment for mitotic indicating that these compounds were similarly active in both cell activity4, oxidative phosphorylation and mitochondrial biogenesis were types (Extended Data Fig. 2l, m). We also found that treatment with the most significant categories to be enriched in F3–T3 human astro- tigecycline (50 mg kg−1) suppressed tumour growth of F3–T3;shTrp53 cytes for each of the three independent comparisons (Fig. 1b, Extended mGSCs glioma xenografts with a more than 50% reduction in tumour 1Institute for Cancer Genetics, Columbia University Medical Center, New York, New York 10032, USA. 2Department of Science and Technology, Universita’ degli Studi del Sannio, Benevento 82100, Italy. 3The Arthur and Sonia Labatt Brain Tumour Research Centre, Program in Developmental and Stem Cell Biology, The Hospital for Sick Children, Toronto, Ontario M5G 1A4, Canada. 4Department of Molecular Genetics, University of Toronto, Toronto, Ontario M5S 1A8, Canada. 5BIOGEM Istituto di Ricerche Genetiche ‘G. Salvatore’, Campo Reale, 83031 Ariano Irpino, Italy. 6Department of Pathology and Laboratory Medicine, Perelman School of Medicine at University of Pennsylvania, Philadelphia, Pennsylvania 19104-6100, USA. 7Qatar Computing Research Institute (QCRI), Hamad Bin Khalifa University, Doha, Qatar. 8Sorbonne Universités UPMC Univ Paris 06, UMR S 1127, Inserm U 1127, CNRS UMR 7225, ICM, Paris 75013, France. 9AP-HP, Groupe Hospitalier Pitié Salpêtrière, Laboratoire de Neuropathologie R Escourolle, Paris 75013, France. 10Onconeurotek, AP-HP, Paris 75013, France. 11AP-HP, Hôpital de la Pitié-Salpêtrière, Service de Neurologie 2, Paris 75013, France. 12Department of Pathology and Cell Biology, Columbia University Medical Center, New York, New York 10032, USA. 13Department of Pediatrics, Columbia University Medical Center, New York, New York 10032, USA. 14Department of Neurology, Columbia University Medical Center, New York, New York 10032, USA. *These authors contributed equally to this work. §These authors jointly supervised this work. a bdc Vector Vector Mitochondrial F3–T3 F3–T3 part Organelle Mitochondrial F3–T3(K508M) F3–T3(K508M) envelope envelope 3.0 ** ** s *** *** Hallmark 500 cell oxidative 2.5 4 phosphorylation 400 Organelle Mitochondrial 2.0 membrane 300 inner ) per 10 Mitochondrion 1.5 3 4 membrane 0 200 2 Mitochondrial 1.0 ×1 0 inner 0.5 100 Envelope membrane mtDNA (fold change) –2 0 P (AU 0 –4 AT F3–T3VF3–T3 ector F3–T3 PD173074 (K508M) Vector DMSO egf Vehicle h F3–T3 DMSO Metformin 1 mM Vehicle Vehicle F3–T3 AZD4547 Tigecycline 10 μM Tigecycline 4 μM Tigecycline (50 mg kg–1) F3–T3 K508M DMSO Menadione 5 μM Tigecycline 8 μM 5 s) 28 ll NS OligogymycinFCCPRotenone NS *** ce 140 NS 1.5 1.2 24 4 NS *** 120 *** 5 1.2 *** * 1 20 6 100 *** (fold change) 6 0.8 16 per 10 ratio 80 ratio 0.9 8 6 –1 me n 0.6 12 60 mi 0.6 8 l volu ** 40 0.4 8 8 ** Survival Survival 8 8 ** ** mo * 20 0.3 0.2 4 8 ** ** P < 0.001 P < 0.001 mour ** 0 0 0 0 020406080 F3–T3 HRAS(12V) Tu 153 7911 OCR (p GSC1123 GSC308 Time (min) Days Figure 1 | Activation of mitochondrial biogenesis and metabolism by in human astrocytes expressing F3–T3, F3–T3(K508M) or vector. d, F3–T3. a, Hierarchical clustering of differentially expressed genes (DEGs) Quantification of cellular ATP in human astrocytes as in c. e, OCR of between F3–T3 human astrocytes, F3–T3 human astrocytes treated with F3–T3 human astrocytes treated with or without AZD4547. f, Survival PD173074 (n = 5 biologically independent samples per group). Human ratio of GSC1123 and GSC308 cells following treatment with the astrocytes expressing the empty vector and F3–T3(K508M) (n = 3 indicated mitochondrial inhibitors. g, Survival ratio of F3–T3;shTrp53 biologically independent samples per group) are included as controls.
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