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Standardization of BCR-ABL p190 molecular analysis

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Descrizione del progetto

BCR-ABL p190 isoform is very frequent in Philadelphia positive (Ph+) Acute Lymphoblastic Leukemia (ALL). The Ph chromosome occurs in about 25% of adult patients with ALL. Prior to the introduction of the BCR-ABL tyrosine kinase inhibitors (TKIs), the outcome of patients with this disease was poor. With the introduction of effective TKIs and immunotherapy, the outcome of patients has significantly improved the choice of transplant is now questionable. It was demonstrated that the minimal residual disease (MRD) status is the most significant risk factor for relapse and monitoring of MRD by quantification of BCR-ABL transcript levels has become a main part of the management of patients with Ph+ ALL. It was also shown that digital (d)PCR is high-sensitive and reliable, allowing an accurate monitoring of BCR-ABL+ ALL patients in complete remission. Furthermore we have preliminary data on Chronic Myeloid Leukemic demonstrating that molecular follow-up by dPCR at several time-points before treatment discontinuation may have a prognostic value for predicting relapse. However neither standard RT-qPCR nor dPCR are standardized for the detection of BCR-ABL p190 isoform. Whereas a lot of work has been done on the standardization of the monitoring of the BCR-ABL p210 isoform in CML, still there is not full agreement on technical approaches, control genes, result expressions, definition of responses and monitoring time-points in Ph+ ALL. I am the coordinator of an Italian group aimed to standardize BCR-ABL p190 monitoring in Ph+ ALL. The group includes 9 well-recognized molecular biology laboratories, including the laboratory at the Department of Clinical and Biological Sciences ,that promoted the standardization of BCR-ABL p210 in Italy and Europe, and are leaders for advanced technologies in ALL both for adults and children. In the first part of our project we sent 3 pooled samples from several ALL patients at different levels of disease to all the participating laboratories, in order to check if results were consistent. Although the data were comparable, they were not overlying, raising the need of a certified reference material to calibrate our analysis. Aim of my project is the production of a certified reference material by three steps in the first year: 1) culture of cell lines SUP-B15 (p190+) and HL60 (p190-); 2) dilution of 4 points of SUP-B15 (p190+) in HL60 (p190-); 3) lyophilization of diluted cells and production of vials for distribution to the Italian laboratories for the titration of the RNA extracted from the cells either by standard PCR and by dPCR. A conversion factor will be produced in the department by statistical analysis of the results then obtained

Risultati e pubblicazioni

Fava C, Rege-Cambrin G, Dogliotti I, Gottardi E, Berchialla P, Di Gioacchino B, Crasto F, Lorenzatti R, Volpengo A, Daraio F, Fantino C, Saglio GEarly BCR-ABL1 Reduction Is Predictive of Better Event-free Survival in Patients With Newly Diagnosed Chronic Myeloid Leukemia Treated With Any Tyrosine Kinase Inhibitor. Clin Lymphoma Myeloma Leuk. 2016;16 Suppl:S96-S100.

Morotti A, Fava CSaglio G. Milestones and monitoring. Curr Hematol Malig Rep. 2015;10:167-72.

Ultimo aggiornamento: 01/11/2017 22:47
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