Tesi di: Giorgia Aimoni Matricola: 707369 Characterization of the genomic breakpoint for molecular monitoring of residual disease in Chronic Myeloid Leukemia Relatore: Prof. Francesco Pasquali Correlatore: Prof. Giovanni Porta Anno Accademico 2010/2011 Università degli Studi dell’Insubria Facoltà di Scienze MM.FF.NN. – Varese Corso di Laurea in Biotecnologie Dipartimento di Scienze Biomediche Sperimentali e Cliniche
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Tesi di: Giorgia AimoniMatricola: 707369
Characterization of the genomic breakpoint
for molecular monitoring of residual disease in
Chronic Myeloid LeukemiaRelatore: Prof. Francesco PasqualiCorrelatore: Prof. Giovanni Porta
Anno Accademico 2010/2011
Università degli Studi dell’InsubriaFacoltà di Scienze MM.FF.NN. – VareseCorso di Laurea in Biotecnologie
Dipartimento di Scienze Biomediche Sperimentali e Cliniche
CHRONIC MYELOID LEUKEMIA
CML is a myeloproliferative disease due to an acquired abnormality that involves the clonal expansion of a hematopoietic stem cell.
1-2 cases per 100,000 person/years, 15% of adult leukemias
DNA quantification (agarose gel and fluorometer) DNA Q-PCR
non-translocated cells leukemic cells
0,0316
0,0010 UND 0,0007 0,0012 0,0002 UND0
0,0050,01
0,0150,02
0,0250,03
0,035
6 42 48 55 58 64 70
Months after Imatinib therapy
% BCR/ABL cells (DNA)
1,82
0 00,234
0 0 00
0,5
1
1,5
2
6 42 48 55 58 64 70Months after Imatinib therapy
Copy number of Bcr/Abl (mRNA)
The discrepancies between the real-time analysis DNA and RNA indicated Ph+ quiescent cells, including leukemic stem cells.
CONCLUSION
Although DNA-based approach requires an initial characterization of the genomic breakpoint, it provides a patient-specific assay that can directly detect the leukemic cells percentage, independent of their state of BCR/ABL expression.
This could help the research and improve the practice of conventional molecular diagnostics.
WORK IN PROGRESS…
DNA Q-PCR analysis on the other patients characterized
Use of KCl22 cell line to validate the method
Ackowledgements
DSBSC - Insubria
Professor Giovanni Porta
Doctor Ilaria Pagani
Professor Francesco Pasquali
Professor Francesco Lo Curto
“Ospedale di Circolo” of Varese and “Ospedali Riuniti” of Bergamo