Detection of EWS chimeric transcripts by nested RT-PCR to allow reinfusion of uncontaminated peripheral blood stem cells in high-risk Ewing's tumor in childhood.
OBJECTIVE
Ewing's tumors (ET) are primary malignancies of bone and soft tissues characterized in at least 96% of cases by specific fusion transcripts originating from recurrent chromosomal translocations. Clinical protocols for high-risk metastatic ETs include high-dose radiation/chemotherapy followed by autologous peripheral blood stem cell (PBSC) reinfusion. We used nested reverse transcriptase polymerase chain reaction (RT-PCR) to search for the presence of ET-specific transcripts in PBSC collections from patients with high-risk ET in order to collect harvests free from neoplastic cells but still sufficient to obtain early stable engraftment.


METHODS
Thirty-seven harvest samples from 15 ET patients treated with mobilizing chemotherapy were analyzed. Nested RT-PCR was performed to detect ET-specific transcripts in RNA extracted from the PBSC collections.


RESULTS
A total of 30 harvests was performed. On average, 2 harvests (range 1-4) were sufficient to collect the minimum required number of mononuclear cells (2.5
Montanaro
L
L
Servizio di Citopatologia; Clinica Pediatrica, Università degli Studi di Bologna, Policlinico S. Orsola, Bologna, Italy.
Pession
A
A
Trerè
D
D
Vici
M
M
Prete
A
A
Paolucci
G
G
Derenzini
M
M
D002648:Child; D002880:Chromosomes, Human, Pair 11; D002892:Chromosomes, Human, Pair 22; D018572:Disease-Free Survival; D018380:Hematopoietic Stem Cell Transplantation / Q000379:methods*; D006801:Humans; D015514:Oncogene Proteins, Fusion / Q000032:analysis* / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000032:analysis* / Q000235:genetics; D012008:Recurrence; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012307:Risk Factors; D012512:Sarcoma, Ewing / Q000097:blood* / Q000453:epidemiology / Q000235:genetics*; D012680:Sensitivity and Specificity; D014158:Transcription, Genetic; D014178:Translocation, Genetic / Q000235:genetics; D014182:Transplantation, Autologous
C122140:EWS-ETV1 fusion protein, human; D015514:Oncogene Proteins, Fusion; D011993:Recombinant Fusion Proteins