Sensitive and Specific Detection of Ewing Sarcoma Minimal Residual Disease in Ovarian and Testicular Tissues in an In Vitro Model

Cancers

Laure Chaput¹  Sandra Sanfilippo¹  Florence Brugnon¹  Pascale Halle²  Bruno Pereira³  Andrei Tchirkov⁴  Lauren Véronèse⁴  Guillaume Martin⁵  Philippe Durand⁵  Nina Radosevic-Robin⁶  Michel Canis⁷  Hervé Lejeune⁸  Justyna Kanold⁹  Victoria Grèze⁹ 

[1] Biologie et Médecine de la Reproduction, AMP (Assistance Médicale à la Procréation), CECOS (Centre d’Etude et de la Conservation des Oeufs et du Sperme), CHU (Centre Hospitalier Universitaire) Estaing, CHU Clermont-Ferrand, 63003 Clermont-Ferrand, France;Centre de Biothérapie d’Auvergne, CHU Estaing, CHU Clermont-Ferrand, 63000 Clermont-Ferrand, France;Délégation à la Recherche Clinique et l’Innovation, Unité de Biostatistiques, CHU Clermont-Ferrand, 63000 Clermont-Ferrand, France;Imagerie Moléculaire et Stratégies Théranostiques Faculté de Médecine, Unité INSERM 1240, Université Clermont Auvergne, 63001 Clermont Ferrand, France;Kallistem, Ecole Normale Supérieure de Lyon, 69007 Lyon, France;Laboratoire de Biologie Médicale (LBM) OncoGenAuvergne, Unité Pathologie, Centre Jean Perrin (CLCC de la région Auvergne), 63011 Clermont-Ferrand, France;Service de Gynécologie, CHU Estaing, CHU Clermont-Ferrand, 63000 Clermont-Ferrand, France;Service de Médecine de la Reproduction, Hôpital Femme Mère Enfant, Hospices Civils de Lyon, 69003 Lyon, France;Service d’Hématologie et d’Oncologie Pédiatrique, CHU Estaing, CHU Clermont-Ferrand, 63000 Clermont-Ferrand, France;

关键词: fertility preservation;    ewing sarcoma;    ovarian tissue;    testicular tissue;    minimal residual disease detection;    rt-qpcr;   

DOI  :  10.3390/cancers11111807

来源: DOAJ

【 摘 要 】

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.

【 授权许可】

Unknown