A genetic model for central chondrosarcoma evolution correlates with patient outcome

Background Central conventional chondrosarcoma (CS) is the most common subtype of primary malignant bone tumour in adults. Treatment options are usually limited to surgery, and prognosis is challenging. These tumours are characterised by the presence and absence of IDH1 and IDH2 mutations, and recently, TERT promoter alterations have been reported in around 20% of cases. The effect of these mutations on clinical outcome remains unclear. The purpose of this study was to determine if prognostic accuracy can be improved by the addition of genomic data, and specifically by examination of IDH1, IDH2, and TERT mutations. Methods In this study, we combined both archival samples and data sourced from the Genomics England 100,000 Genomes Project (n = 356). Mutations in IDH1, IDH2, and TERT were profiled using digital droplet PCR (n = 346), whole genome sequencing (n=68), or both (n = 64). Complex events and other genetic features were also examined, along with methylation array data (n = 84). We correlated clinical features and patient outcomes with our genetic findings. Results IDH2-mutant tumours occur in older patients and commonly present with high-grade or dedifferentiated disease. Notably, TERT mutations occur most frequently in IDH2-mutant tumours, although have no effect on survival in this group. In contrast, TERT mutations are rarer in IDH1-mutant tumours, yet they are associated with a less favourable outcome in this group. We also found that methylation profiles distinguish IDH1- from IDH2-mutant tumours. IDH wild-type tumours rarely exhibit TERT mutations and tend to be diagnosed in a younger population than those with tumours harbouring IDH1 and IDH2 mutations. A major genetic feature of this group is haploidisation and subsequent genome doubling. These tumours evolve less frequently to dedifferentiated disease and therefore constitute a lower risk group. Conclusions Tumours with IDH1 or IDH2 mutations or those that are IDHwt have significantly different genetic pathways and outcomes in relation to TERT mutation. Diagnostic testing for IDH1, IDH2, and TERT mutations could therefore help to guide clinical monitoring and prognostication

Cancer origin tracing and timing in two high-risk prostate cancers using multisample whole genome analysis: prospects for personalized medicine

BACKGROUND: Prostate cancer (PrCa) genomic heterogeneity causes resistance to therapies such as androgen deprivation. Such heterogeneity can be deciphered in the context of evolutionary principles, but current clinical trials do not include evolution as an essential feature. Whether or not analysis of genomic data in an evolutionary context in primary prostate cancer can provide unique added value in the research and clinical domains remains an open question. METHODS: We used novel processing techniques to obtain whole genome data together with 3D anatomic and histomorphologic analysis in two men (GP5 and GP12) with high-risk PrCa undergoing radical prostatectomy. A total of 22 whole genome-sequenced sites (16 primary cancer foci and 6 lymph node metastatic) were analyzed using evolutionary reconstruction tools and spatio-evolutionary models. Probability models were used to trace spatial and chronological origins of the primary tumor and metastases, chart their genetic drivers, and distinguish metastatic and non-metastatic subclones. RESULTS: In patient GP5, CDK12 inactivation was among the first mutations, leading to a PrCa tandem duplicator phenotype and initiating the cancer around age 50, followed by rapid cancer evolution after age 57, and metastasis around age 59, 5 years prior to prostatectomy. In patient GP12, accelerated cancer progression was detected after age 54, and metastasis occurred around age 56, 3 years prior to prostatectomy. Multiple metastasis-originating events were identified in each patient and tracked anatomically. Metastasis from prostate to lymph nodes occurred strictly ipsilaterally in all 12 detected events. In this pilot, metastatic subclone content analysis appears to substantially enhance the identification of key drivers. Evolutionary analysis' potential impact on therapy selection appears positive in these pilot cases. CONCLUSIONS: PrCa evolutionary analysis allows tracking of anatomic site of origin, timing of cancer origin and spread, and distinction of metastatic-capable from non-metastatic subclones. This enables better identification of actionable targets for therapy. If extended to larger cohorts, it appears likely that similar analyses could add substantial biological insight and clinically relevant value

Genomic and transcriptomic characterisation of undifferentiated pleomorphic sarcoma of bone

This is an accepted manuscript of an article published by Wiely in The Journal of Pathology on 27/12/2018, available online: https://doi.org/10.1002/path.5176 The accepted version of the publication may differ from the final published version.Undifferentiated pleomorphic sarcoma of bone (UPSb), is a rare primary bone sarcoma that lacks a specific line of differentiation. There is very little information about the genetic alterations leading to tumourigenesis or malignant transformation. Distinguishing between UPSb and other malignant bone sarcomas, including dedifferentiated chondrosarcoma and osteosarcoma, can be challenging due to overlapping features. To explore the genomic and transcriptomic landscape of UPSb tumours, whole-exome sequencing (WES) and RNA Sequencing (RNA-Seq) were performed on UPSb tumours. All tumours lacked hotspot mutations in IDH1/2 132 or 172 codons, thereby excluding the diagnosis of dedifferentiated chondrosarcoma. Recurrent somatic mutations in TP53 were identified in 4/14 samples (29%). Moreover, recurrent mutations in histone chromatin remodelling genes, including H3F3A, ATRX and DOT1L, were identified in 5/14 samples (36%), highlighting the potential role of deregulated chromatin remodelling pathways in UPSb tumourigenesis. The majority of recurrent mutations in chromatin remodelling genes identified here are reported in COSMIC, including the H3F3A G35 and K36 hotspot residues. Copy number alteration analysis identified gains and losses in genes that have been previously altered in UPSb or UPS of soft tissue. Eight somatic gene fusions were identified by RNA-Seq, two of which, CLTC-VMP1 and FARP1-STK24, were reported previously in multiple cancers. Five gene fusions were genomically characterised. Hierarchical clustering analysis, using RNA-Seq data, distinctly clustered UPSb tumours from osteosarcoma and other sarcomas, thus molecularly distinguishing UPSb from other sarcomas. RNA-Seq expression profiling analysis and quantitative RT-PCR showed an elevated expression in FGF23 which can be a potential molecular biomarker in UPSb. To our knowledge, this study represents the first comprehensive WES and RNA-Seq analysis of UPSb tumours revealing novel protein-coding recurrent gene mutations, gene fusions and identifying a potential UPSb molecular biomarker, thereby broadening the understanding of the pathogenic mechanisms and highlighting the possibility of developing novel targeted therapeutics

Transcription factor motif quality assessment requires systematic comparative analysis [version 2; referees: 2 approved]

Transcription factor (TF) binding site prediction remains a challenge in gene regulatory research due to degeneracy and potential variability in binding sites in the genome. Dozens of algorithms designed to learn binding models (motifs) have generated many motifs available in research papers with a subset making it to databases like JASPAR, UniPROBE and Transfac. The presence of many versions of motifs from the various databases for a single TF and the lack of a standardized assessment technique makes it difficult for biologists to make an appropriate choice of binding model and for algorithm developers to benchmark, test and improve on their models. In this study, we review and evaluate the approaches in use, highlight differences and demonstrate the difficulty of defining a standardized motif assessment approach. We review scoring functions, motif length, test data and the type of performance metrics used in prior studies as some of the factors that influence the outcome of a motif assessment. We show that the scoring functions and statistics used in motif assessment influence ranking of motifs in a TF-specific manner. We also show that TF binding specificity can vary by source of genomic binding data. We also demonstrate that information content of a motif is not in isolation a measure of motif quality but is influenced by TF binding behaviour. We conclude that there is a need for an easy-to-use tool that presents all available evidence for a comparative analysis

Remodelage génomique des sarcomes pléomorphes : caractérisation transcriptomique et agressivité tumorale

Pleomorphic sarcomas are rare mesenchymal tumors characterized by many chromosomal rearrangements. Their oncogenic process is still poorly understood, no recurrent and specific genetic alteration able to drive the tumor initiation has been identified yet. The work I did during my thesis had the objective to better understand the biology of these tumors, focusing on transcriptomic consequences of their genomic remodeling.We characterized fusion transcripts in these tumors by high-throughput sequencing (RNA-seq). This led us to identify the expression of several chimeric transcripts involving TRIO (5.1% of cases). Moreover, this analysis and the identification of expressed variants allowed us to identify frequent mutations of tumor suppressor genes such as ATRX (16% of cases) and more generally members of the SWI/SNF complex (47% of cases). Alterations of this major complex of chromatin remodeling are associated with higher genetic instability and more aggressive phenotype.In pleomorphic sarcomas, genetic instability is linked to tumor progression through the expression of a prognostic transcriptomic signature. This signature, termed CINSARC, is involved in mitosis control and chromosome segregation pathways. We wanted to determine the origin of such expression by integrating genomics and epigenetics, transcriptional and post-transcriptional regulation mechanisms. Though these mechanisms do not seem to directly regulate CINSARC expression, important changes have been highlighted. From a clinical standpoint, we demonstrated that the signature expression is a global prognostic factor of tumor aggressiveness in numerous cancer types. In addition, CINSARC is a better prognostic marker than the FNCLCC grading system, the current international standard to evaluate the metastatic risk in soft tissue sarcomas. We consequently developed a method allowing a daily clinical application of the CINSARC signature to improve the therapeutic management of these tumors.Les sarcomes pléomorphes sont des tumeurs mésenchymateuses rares caractérisées par de nombreux remaniements chromosomiques. Leur processus d’oncogenèse reste encore mal compris, aucune altération génétique motrice de l’initiation tumorale n’a pu être identifiée de façon récurrente et spécifique à ce jour. Les travaux que j’ai réalisés durant ma thèse avaient pour but de mieux comprendre la biologie de ces tumeurs, notamment les conséquences transcriptomiques de leur remodelage génomique.Nous avons caractérisé les transcrits de fusion exprimés dans ces tumeurs par séquençage haut-débit (RNA-seq). Ceci nous a amené à identifier l’expression de plusieurs transcrits chimériques impliquant le gène TRIO (5,1% des tumeurs). De plus, cette analyse ainsi que l’identification de variants exprimés nous ont permis d’identifier de fréquentes mutations de gènes suppresseurs de tumeurs tels qu’ATRX (16% des tumeurs) et plus généralement des membres du complexe SWI/SNF (47% des tumeurs). Les altérations de ce complexe majeur de remodelage de la chromatine sont associées à une plus grande instabilité génétique et à un phénotype plus agressif.Dans les sarcomes pléomorphes, l’instabilité génétique est liée à la progression tumorale via l’expression d’une signature transcriptomique pronostique. Cette signature, nommée CINSARC, est impliquée dans le contrôle de la mitose et de la ségrégation chromosomique. Nous avons voulu déterminer l’origine de cette expression via une étude intégrant la génomique et des mécanismes de régulation épigénétique, transcriptionnelle et post-transcriptionnelle. Si ces mécanismes ne semblent pas directement causals de l’expression de CINSARC, d’importantes modifications ont pu être mises en évidences. D’un point de vue clinique, nous avons démontré que l’expression de cette signature est un facteur pronostique universel de l’agressivité tumorale dans de nombreux types de cancers. De plus, CINSARC est un meilleur marqueur pronostique que le grade FNCLCC, actuel standard international d’évaluation du risque métastatique des sarcomes des tissus mous. Nous avons ainsi développé une méthode permettant une application clinique routinière de la signature CINSARC afin d’améliorer la prise en charge thérapeutique de ces tumeurs

Genomic remodeling of pleomorphic sarcomas : transcriptomic characterization and tumor aggressiveness

Les sarcomes pléomorphes sont des tumeurs mésenchymateuses rares caractérisées par de nombreux remaniements chromosomiques. Leur processus d’oncogenèse reste encore mal compris, aucune altération génétique motrice de l’initiation tumorale n’a pu être identifiée de façon récurrente et spécifique à ce jour. Les travaux que j’ai réalisés durant ma thèse avaient pour but de mieux comprendre la biologie de ces tumeurs, notamment les conséquences transcriptomiques de leur remodelage génomique.Nous avons caractérisé les transcrits de fusion exprimés dans ces tumeurs par séquençage haut-débit (RNA-seq). Ceci nous a amené à identifier l’expression de plusieurs transcrits chimériques impliquant le gène TRIO (5,1% des tumeurs). De plus, cette analyse ainsi que l’identification de variants exprimés nous ont permis d’identifier de fréquentes mutations de gènes suppresseurs de tumeurs tels qu’ATRX (16% des tumeurs) et plus généralement des membres du complexe SWI/SNF (47% des tumeurs). Les altérations de ce complexe majeur de remodelage de la chromatine sont associées à une plus grande instabilité génétique et à un phénotype plus agressif.Dans les sarcomes pléomorphes, l’instabilité génétique est liée à la progression tumorale via l’expression d’une signature transcriptomique pronostique. Cette signature, nommée CINSARC, est impliquée dans le contrôle de la mitose et de la ségrégation chromosomique. Nous avons voulu déterminer l’origine de cette expression via une étude intégrant la génomique et des mécanismes de régulation épigénétique, transcriptionnelle et post-transcriptionnelle. Si ces mécanismes ne semblent pas directement causals de l’expression de CINSARC, d’importantes modifications ont pu être mises en évidences. D’un point de vue clinique, nous avons démontré que l’expression de cette signature est un facteur pronostique universel de l’agressivité tumorale dans de nombreux types de cancers. De plus, CINSARC est un meilleur marqueur pronostique que le grade FNCLCC, actuel standard international d’évaluation du risque métastatique des sarcomes des tissus mous. Nous avons ainsi développé une méthode permettant une application clinique routinière de la signature CINSARC afin d’améliorer la prise en charge thérapeutique de ces tumeurs.Pleomorphic sarcomas are rare mesenchymal tumors characterized by many chromosomal rearrangements. Their oncogenic process is still poorly understood, no recurrent and specific genetic alteration able to drive the tumor initiation has been identified yet. The work I did during my thesis had the objective to better understand the biology of these tumors, focusing on transcriptomic consequences of their genomic remodeling.We characterized fusion transcripts in these tumors by high-throughput sequencing (RNA-seq). This led us to identify the expression of several chimeric transcripts involving TRIO (5.1% of cases). Moreover, this analysis and the identification of expressed variants allowed us to identify frequent mutations of tumor suppressor genes such as ATRX (16% of cases) and more generally members of the SWI/SNF complex (47% of cases). Alterations of this major complex of chromatin remodeling are associated with higher genetic instability and more aggressive phenotype.In pleomorphic sarcomas, genetic instability is linked to tumor progression through the expression of a prognostic transcriptomic signature. This signature, termed CINSARC, is involved in mitosis control and chromosome segregation pathways. We wanted to determine the origin of such expression by integrating genomics and epigenetics, transcriptional and post-transcriptional regulation mechanisms. Though these mechanisms do not seem to directly regulate CINSARC expression, important changes have been highlighted. From a clinical standpoint, we demonstrated that the signature expression is a global prognostic factor of tumor aggressiveness in numerous cancer types. In addition, CINSARC is a better prognostic marker than the FNCLCC grading system, the current international standard to evaluate the metastatic risk in soft tissue sarcomas. We consequently developed a method allowing a daily clinical application of the CINSARC signature to improve the therapeutic management of these tumors

Genomic remodeling of pleomorphic sarcomas : transcriptomic characterization and tumor aggressiveness

Les sarcomes pléomorphes sont des tumeurs mésenchymateuses rares caractérisées par de nombreux remaniements chromosomiques. Leur processus d’oncogenèse reste encore mal compris, aucune altération génétique motrice de l’initiation tumorale n’a pu être identifiée de façon récurrente et spécifique à ce jour. Les travaux que j’ai réalisés durant ma thèse avaient pour but de mieux comprendre la biologie de ces tumeurs, notamment les conséquences transcriptomiques de leur remodelage génomique.Nous avons caractérisé les transcrits de fusion exprimés dans ces tumeurs par séquençage haut-débit (RNA-seq). Ceci nous a amené à identifier l’expression de plusieurs transcrits chimériques impliquant le gène TRIO (5,1% des tumeurs). De plus, cette analyse ainsi que l’identification de variants exprimés nous ont permis d’identifier de fréquentes mutations de gènes suppresseurs de tumeurs tels qu’ATRX (16% des tumeurs) et plus généralement des membres du complexe SWI/SNF (47% des tumeurs). Les altérations de ce complexe majeur de remodelage de la chromatine sont associées à une plus grande instabilité génétique et à un phénotype plus agressif.Dans les sarcomes pléomorphes, l’instabilité génétique est liée à la progression tumorale via l’expression d’une signature transcriptomique pronostique. Cette signature, nommée CINSARC, est impliquée dans le contrôle de la mitose et de la ségrégation chromosomique. Nous avons voulu déterminer l’origine de cette expression via une étude intégrant la génomique et des mécanismes de régulation épigénétique, transcriptionnelle et post-transcriptionnelle. Si ces mécanismes ne semblent pas directement causals de l’expression de CINSARC, d’importantes modifications ont pu être mises en évidences. D’un point de vue clinique, nous avons démontré que l’expression de cette signature est un facteur pronostique universel de l’agressivité tumorale dans de nombreux types de cancers. De plus, CINSARC est un meilleur marqueur pronostique que le grade FNCLCC, actuel standard international d’évaluation du risque métastatique des sarcomes des tissus mous. Nous avons ainsi développé une méthode permettant une application clinique routinière de la signature CINSARC afin d’améliorer la prise en charge thérapeutique de ces tumeurs.Pleomorphic sarcomas are rare mesenchymal tumors characterized by many chromosomal rearrangements. Their oncogenic process is still poorly understood, no recurrent and specific genetic alteration able to drive the tumor initiation has been identified yet. The work I did during my thesis had the objective to better understand the biology of these tumors, focusing on transcriptomic consequences of their genomic remodeling.We characterized fusion transcripts in these tumors by high-throughput sequencing (RNA-seq). This led us to identify the expression of several chimeric transcripts involving TRIO (5.1% of cases). Moreover, this analysis and the identification of expressed variants allowed us to identify frequent mutations of tumor suppressor genes such as ATRX (16% of cases) and more generally members of the SWI/SNF complex (47% of cases). Alterations of this major complex of chromatin remodeling are associated with higher genetic instability and more aggressive phenotype.In pleomorphic sarcomas, genetic instability is linked to tumor progression through the expression of a prognostic transcriptomic signature. This signature, termed CINSARC, is involved in mitosis control and chromosome segregation pathways. We wanted to determine the origin of such expression by integrating genomics and epigenetics, transcriptional and post-transcriptional regulation mechanisms. Though these mechanisms do not seem to directly regulate CINSARC expression, important changes have been highlighted. From a clinical standpoint, we demonstrated that the signature expression is a global prognostic factor of tumor aggressiveness in numerous cancer types. In addition, CINSARC is a better prognostic marker than the FNCLCC grading system, the current international standard to evaluate the metastatic risk in soft tissue sarcomas. We consequently developed a method allowing a daily clinical application of the CINSARC signature to improve the therapeutic management of these tumors