• Circulating tumour cells
• metastasis
• soft tissue sarcoma

• diagnostic markers
• predictive markers
• prognostic markers
• paediatric cancer

• Stichting Kinderen Kankervrij (KiKa)

• biopsy
• incisional
• liquid
• percutaneous

• Humans
• Biopsy, Needle/methods
• Reproducibility of Results
• Bone Neoplasms
• Biopsy
• Soft Tissue Neoplasms
• Musculoskeletal Diseases

• Humans
• Male
• Adolescent
• Young Adult
• Adult
• Middle Aged
• Female
• Sarcoma, Ewing/diagnostic imaging
• Sarcoma, Ewing/pathology
• Bone Neoplasms/pathology
• Positron Emission Tomography Computed Tomography/methods
• Bone Marrow/pathology
• Retrospective Studies
• Clinical Relevance
• Biopsy
• Tomography, X-Ray Computed
• Neoplasm Staging

• Ewing’s sarcoma
• angiosarcoma
• chemotherapy
• osteosarcoma
• radiotherapy
• rhabdomyosarcoma

• Humans
• Sarcoma/pathology
• Head and Neck Neoplasms/therapy
• Soft Tissue Neoplasms/pathology
• Neck/pathology

• 2020-J-3 The National Cancer Center Research and Development Fund

• RRBS
• cfDNA (cell-free DNA)
• ddPCR
• liquid biopsy
• mRNA
• pediatric cancer
• qPCR
• solid tumor

• CTC
• Ewing's sarcoma
• Osteosarcoma
• Rhabdomyosarcoma
• ctDNA
• ctRNA

Ewing sarcoma (ES) rarely derives from the sacrum or mobile spine. The discovery of primary ES with multimetastatic involvements is exceedingly less frequent in clinical practice. A 23-year-old man with initial primary sacral ES developed metastases of rib, lung, and multifocal skull after receiving surgical intervention and series of adjuvant therapies. We provide this very rare case consisting of its clinical features, imaging findings, treatments, and outcomes. Therapeutic modalities of ES are also reviewed in previous published articles. The prognosis of metastatic ES remains dismal; effective therapeutic modalities for ES require multidisciplinary collaboration, with more high-quality clinical trials to promote the optimal protocols.

• sarcoma
• cancer genetics

• CD99+CD45−
• Ewing sarcoma
• flow cytometry
• peripheral blood

• Cannabidiol
• Cannabinoid receptors
• Cell viability
• Cytotoxicity
• Ewing sarcoma
• WIN-55,212-2

Minimal residual disease (MRD) measurement is a cornerstone of contemporary acute lymphoblastic leukaemia (ALL) treatment. The presence of immunoglobulin (Ig) and T cell receptor (TCR) gene recombinations in leukaemic clones allows widespread use of patient-specific, DNA-based MRD assays. In contrast, paediatric solid tumour MRD remains experimental and has focussed on generic assays targeting tumour-specific messenger RNA, methylated DNA or microRNA.

We examined the feasibility of using whole-genome sequencing (WGS) data to design tumour-specific polymerase chain reaction (PCR)-based MRD tests (WGS-MRD) in 18 children with high-risk relapsed cancer, including ALL, high-risk neuroblastoma (HR-NB) and Ewing sarcoma (EWS) (n = 6 each).

Sensitive WGS-MRD assays were generated for each patient and allowed quantitation of 1 tumour cell per 10⁻⁴ (0.01%)–10^–5 (0.001%) mononuclear cells. In ALL, WGS-MRD and Ig/TCR-MRD were highly concordant. WGS-MRD assays also showed good concordance between quantitative PCR and droplet digital PCR formats. In serial clinical samples, WGS-MRD correlated with disease course. In solid tumours, WGS-MRD assays were more sensitive than RNA-MRD assays.

WGS facilitated the development of patient-specific MRD tests in ALL, HR-NB and EWS with potential clinical utility in monitoring treatment response. WGS data could be used to design patient-specific MRD assays in a broad range of tumours.

The present review is aimed to discuss the relevance of assaying for the presence and isolation of circulating tumor cells (CTCs) in patients with sarcoma. Just a few studies have been performed to detect and enumerate viable CTCs in sarcoma and a majority of them still represent proof-of-concept studies, while more frequently tumor cells have been detected in the circulation by using the PCR-based method. Nevertheless, recent advances in technologies allowed detection of epithelial–mesenchymal transitioned CTCs from patients with mesenchymal malignancies, despite results being mostly preliminary. The possibility to identify CTCs holds a great promise for both applications of liquid biopsy in sarcoma for precision medicine, and for research purposes to pinpoint the mechanism of the metastatic process through the characterization of tumor mesenchymal cells. Coherently, clinical trials in sarcoma have been designed accordingly to detect CTCs, for diagnosis, identification of novel therapeutic targets and resistance mechanisms of systemic therapies, and patient stratification.

Bone and soft tissue sarcomas (STSs) represent a group of heterogeneous rare malignant tumors of mesenchymal origin, with a poor prognosis. Due to their low incidence, only a few studies have been reported addressing circulating tumor cells (CTCs) in sarcoma, despite the well-documented relevance for applications of liquid biopsy in precision medicine. In the present review, the most recent data relative to the detection and isolation of viable and intact CTCs in these tumors will be reviewed, and the heterogeneity in CTCs will be discussed. The relevance of epithelial–mesenchymal plasticity and stemness in defining the phenotypic and functional properties of these rare cells in sarcoma will be highlighted. Of note, the existence of dynamic epithelial–mesenchymal transition (EMT)-related processes in sarcoma tumors has only recently been related to their clinical aggressiveness. Also, the presence of epithelial cell adhesion molecule (EpCAM)-positive CTC in sarcoma has been weakly correlated with poor outcome and disease progression, thus proving the existence of both epithelial and mesenchymal CTC in sarcoma. The advancement in technologies for capturing and enumerating all diverse CTCs phenotype originating from these mesenchymal tumors are presented, and results provide a promising basis for clinical application of CTC detection in sarcoma.

• CTC
• clinical practice
• epithelial–mesenchymal plasticity
• sarcoma

• children and adolescents
• clinical application
• minimal disseminated disease
• minimal residual disease
• non-Hodgkin lymphoma

• Ewing sarcoma
• NK cells
• NSG mice
• PuMA
• lung metastasis
• osteosarcoma

• extraskeletal ewing sarcoma
• multidisciplinary treatment
• penis

• Adult
• Humans
• Male
• Penile Neoplasms/pathology
• Penis/pathology
• Sarcoma, Ewing/pathology

• EWSR1
• biomarkers
• early detection
• liquid biopsy
• sarcoma

• 30309 Cancer Research UK
• 68105/A30309 Cancer Research UK

Ewing sarcoma (EWS) is a paediatric aggressive malignant tumour of bones and soft tissues. Multidisciplinary chemotherapies, surgical resection, and radiation represent the only strategies counteracting the disease, however spreading and relapse of disease still remain a clinical issue. Circulating tumour cells (CTCs) are an important feature of EWS but the prognostic significance has not been, yet, clarified. CTCs have been found both in patients with localized disease and in those who recur or metastasize. The identification of markers that can detect recurrences and metastasis remains an important challenge for research. Unfortunately, even most of patients with localized cancer relapsed and the reason has not yet been fully understood. In this clinical study on EWS patients, we evaluated the expression of CD99 antigen and beta-3 adrenergic receptor (β3-AR) on CTCs and bioptic derived cells by flow cytometry. The preliminary data revealed a higher β3-AR expression on cells derived from metastatic or relapsed patients, suggesting a role for the β3-AR as a possible predictive maker of disease recurrence in both patients with metastatic and localized disease.

• circulating tumour cells
• ewing sarcoma
• β3-adrenergic receptors

• Circulating biomarkers
• Ewing sarcoma
• Metastases
• miR34a
• miRNAs

• IG 18451 Associazione Italiana per la Ricerca sul Cancro
• fellowship "SITE" 19498 Associazione Italiana per la Ricerca sul Cancro
• PER-2011-2353839 PROVABES

• Ewing sarcoma
• acquisition of self-renewal traits
• aggressiveness
• chemoresistance
• clinical pathogenesis
• therapeutic target
• tyrosine kinases

• Cell-free DNA
• Epigenetics
• Liquid biopsy
• Next-generation sequencing
• Pediatric solid tumors

Ewing sarcoma (EWS) is a common pediatric solid tumor with high metastatic potential. Due to toxic effects of treatments on reproductive functions, the cryopreservation of ovarian tissue (OT) or testicular tissue (TT) is recommended to preserve fertility. However, the risk of reintroducing residual metastatic tumor cells should be evaluated before fertility restoration. Our goal was to validate a sensitive and specific approach for EWS minimal residual disease (MRD) detection in frozen germinal tissues. Thawed OT (n = 12) and TT (n = 14) were contaminated with tumor RD-ES cells (10, 100, and 1000 cells) and EWS-FLI1 tumor-specific transcript was quantified with RT-qPCR. All contaminated samples were found to be positive, with a strong correlation between RD-ES cell numbers and EWS-FLI1 levels in OT (r = 0.93) and TT (r = 0.96) (p < 0.001). No transcript was detected in uncontaminated control samples. The invasive potential of Ewing cells was evaluated using co-culture techniques. After co-culturing, tumor cells were detected in OT/TT with histology, FISH, and RT-qPCR. In addition, four OT and four TT samples from children with metastatic EWS were tested, and no MRD was found using RT-qPCR and histology. We demonstrated the high sensitivity and specificity of RT-qPCR to detect EWS MRD in OT/TT samples. Clinical trial: NCT 02400970.

• Ewing sarcoma
• RT-qPCR
• fertility preservation
• minimal residual disease detection
• ovarian tissue
• testicular tissue

• Ewing sarcoma
• bone marrow biopsy
• staging

• Droplet digital PCR
• Liquid biopsy
• Nested PCR
• SS18-SSX fusion transcript
• Synovial sarcoma

• Bone and soft tissue sarcomas
• Circulating cell-free nucleic acids
• Circulating tumor cells
• Exosomes
• Metabolites

• Biomarkers, Tumor/blood
• Biomarkers, Tumor/genetics
• Bone Neoplasms/pathology
• Bone Neoplasms/therapy
• Bone and Bones/metabolism
• Bone and Bones/pathology
• Cell-Free Nucleic Acids/blood
• Cell-Free Nucleic Acids/genetics
• Exosomes/metabolism
• Forecasting
• Humans
• Liquid Biopsy/methods
• Liquid Biopsy/trends
• Neoplastic Cells, Circulating/metabolism
• Sarcoma/pathology
• Sarcoma/therapy
• Soft Tissue Neoplasms/pathology
• Soft Tissue Neoplasms/therapy

• U01 CA151452 NCI NIH HHS

• Ewing sarcoma
• anti-CD99 antibody
• circulating tumor cells
• immunoseparation
• liquid biopsy
• magnetic beads

• gemcitabine
• docetaxel
• ewing sarcoma
• adolescent and young adult sarcomas
• minimal residual disease management

• Adolescent
• Adult
• Age Factors
• Antineoplastic Combined Chemotherapy Protocols/therapeutic use
• Bone Neoplasms/drug therapy
• Child
• Child, Preschool
• Deoxycytidine/administration & dosage
• Deoxycytidine/analogs & derivatives
• Disease-Free Survival
• Docetaxel
• Humans
• Kaplan-Meier Estimate
• Odds Ratio
• Prognosis
• Prospective Studies
• Sarcoma, Ewing/drug therapy
• Sarcoma, Ewing/mortality
• Spain
• Survival Rate
• Taxoids/administration & dosage
• Gemcitabine

• Blood
• Bone marrow
• Circulating
• Disseminating
• Neuroblastoma
• RTqPCR
• Tumour cells

• 18226 Cancer Research UK

Sarcomas are a heterogeneous group of malignant neoplasms of mesenchymal origin, many of which have a propensity to develop distant metastases. Cancer cells that have escaped from the primary tumor are able to invade into surrounding tissues, to intravasate into the bloodstream to become circulating tumor cells (CTCs), and are responsible for the generation of distant metastases. Due to the rarity of these tumors and the absence of specific markers expressed by sarcoma tumor cells, the characterization of sarcoma CTCs has to date been relatively limited. Current techniques for isolating sarcoma CTCs are based on size criteria, the identification of circulating cells that express either common mesenchymal markers, sarcoma-specific markers, such as CD99, CD81, or PAX3, and chromosomal translocations found in certain sarcoma subtypes, such as EWS-FLI1 in Ewing’s sarcoma, detection of osteoblast-related genes, or measurement of the activity of specific metabolic enzymes. Further studies are needed to improve the isolation and characterization of sarcoma CTCs, to demonstrate their clinical significance as predictive and/or prognostic biomarkers, and to utilize CTCs as a tool for investigating the metastatic process in sarcoma and to identify novel therapeutic targets. The present review provides a short overview of the most recent literature on CTCs in sarcoma.

• cancer stem cells
• circulating tumor cells
• neuroblastoma
• rare cancers
• sarcoma

• Ewing sarcoma
• fertility preservation
• malignant cell detection
• minimal disseminated disease
• molecular markers
• ovarian metastasis
• ovarian tissue cryopreservation
• rhabdomyosarcoma

• anti-müllerian hormone
• ewing sarcoma
• female fertility
• osteosarcoma
• ovarian reserve
• wilms tumor

• Adolescent
• Anti-Mullerian Hormone/blood
• Bone Neoplasms/complications
• Bone Neoplasms/therapy
• Child
• Child, Preschool
• Combined Modality Therapy/adverse effects
• Combined Modality Therapy/methods
• Female
• Follicle Stimulating Hormone/blood
• Humans
• Infant
• Kidney Neoplasms/complications
• Kidney Neoplasms/therapy
• Luteinizing Hormone/blood
• Osteosarcoma/complications
• Osteosarcoma/therapy
• Primary Ovarian Insufficiency/etiology
• Sarcoma, Ewing/complications
• Sarcoma, Ewing/therapy
• Wilms Tumor/complications
• Wilms Tumor/therapy

Objective: To outline the epidemiological profile and prognosis for Ewing's sarcoma in the Brazilian population. Material and Methods: The medical records of 64 patients with intraosseous Ewing's sarcoma who were treated at the Pediatric Oncology Institute, IOP-GRAACC-Unifesp, between 1995 and 2010, were retrospectively evaluated. Results: The statistical analysis on the data obtained did not correlate factors such as sex, trauma, pathological fracture and time taken for case diagnosis with the treatment outcome. Factors such as initial metastasis, lung metastasis, tumor site, age, recurrence and type of surgery showed results corroborating what has been established in the literature. Conclusion: The prognosis in cases of Ewing's sarcoma was mainly influenced by the presence of metastases at the time of diagnosis.

• Bone Neoplasms
• Prognosis
• Sarcoma, Ewing

• Ewing sarcoma
• bone marrow aspiration
• bone marrow biopsy
• bone scan
• pediatrics
• staging