A subset of epithelioid and spindle cell rhabdomyosarcomas is associated with TFCP2 fusions and common ALK upregulation


Rhabdomyosarcomas with TFCP2 fusions represent an emerging subtype of tumors, initially discovered by RNA-sequencing. We report herein the clinicopathological, transcriptional, and genomic features of a series of 14 cases. Cases were retrospectively and prospectively recruited and studied by immunohistochemistry (MYF4, MYOD1, S100, AE1/E3, ALK), fluorescence in situ hybridization with TFCP2 break-apart probe (n = 10/14), array-comparative genomic hybridization (Agilent), whole RNA-sequencing (Truseq Exome, Illumina), or anchored multiplex PCR-based targeted next-generation sequencing (Archer® FusionPlex® Sarcoma kit). Patient’s age ranged between 11 and 86 years, including 5 pediatric cases. Tumors were located in the bone (n = 12/14) and soft tissue (n = 2/14). Most bone tumors invaded surrounding soft tissue. Craniofacial bones were over-represented (n = 8/12). Median survival was 8 months and five patients are currently alive with a median follow-up of 20 months. Most tumors displayed a mixed spindle cell and epithelioid pattern with frequent vesicular nuclei. All tumors expressed keratins and showed a rhabdomyogenic phenotype (defined as expression of MYF4 and/or MYOD1). ALK was overexpressed in all but three cases without underlying ALK fusion on break-apart FISH (n = 5) nor next-generation sequencing (n = 14). ALK upregulation was frequently associated with an internal deletion at genomic level. TFCP2 was fused in 5′ either to EWSR1 (n = 6) or FUS (n = 8). EWSR1 was involved in both soft tissue cases. FISH with TFCP2 break-apart probe was positive in all tested cases (n = 8), including one case with unbalanced signal. On array-CGH, all tested tumors displayed complex genetic profiles with genomic indexes ranging from 13 to 107.55 and recurrent CDKN2A deletions. FET-TFCP2 rhabdomyosarcomas clustered together and distinctly from other rhabdomyosarcomas subgroups. Altogether, our data confirm and expand the spectrum of the new family of FET-TFCP2 rhabdomyosarcomas, which are associated with a predilection for the craniofacial bones, an aggressive course, and recurrent pathological features. Their association with ALK overexpression might represent a therapeutic vulnerability.

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We are grateful to pathologists and clinicians for sharing cases and follow-up informations: Dr A Al Bouzidi (Rabat, Marocco), Dr E Angot (CHU Rouen), Dr Foucard (Paris), Professor C Berger (HEGP, Paris), Dr N Bennani Guebessi (Casablanca, Marocco), Dr C Bazille (CHU Caen), Professor F Gouin (CHU Nantes), Dr E Bompas (ICO, St Herblain), Dr Haffadi (Casablanca), Dr C Raban (CHU Poitiers), Dr V Laurence (Institut Curie, Paris), Dr A Moreau (CHU Nantes, France), Dr S Watson (Institut Curie, Paris, France), Professor D Baumhoer (Basel, Switzerland), Dr M Chan (Paramaribo, Suriname), and Dr U Flucke (Nijmegen, The Netherlands). Dr A de La Fouchardière (Centre Leon Berard, Lyon, France) graciously provided the ALK-fusion positive Spitz nevi for the transcriptional analysis. We thank L Mayeur, V Velasco, and V Fouqué for their assistance. INSERM U1218 is supported by a charity funding “Le Fil d’Oriane”.

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