Study of chromosomal abnormalities by fluorescent in-situ-hyberdisation (FISH), and quantitative polymerase chain reaction (qPCR) in Multiple Myeloma patients and correlations with treatment outcomes.

Multiple Myeloma (MM) is a plasma cell dyscrasia resulting in high levels of monoclonal immunoglobulin (Ig) production leading to immunosuppression and other physiological disturbances. A median survival of about 2 years is usually obtained with conventional chemotherapy, however trials using high-dose therapy with subsequent autologous stem cell rescue have improved the remission rate, event-free survival, and overall survival (OS) in certain cohorts of patients. Various chromosomal abnormalities have been reported in MM patients but the 13q- and t(4:14) abnormalities are present in over 50% of abnormal plasma cells in these patients though the clinical significance of this has not been elucidated. Reports have linked the presence of the 13q- abnormality with altered therapeutic responses to common treatments. Strong evidence now indicates that conventional cytogenetics fails to recognise about 50% of these abnormalities because of slow growth of MM cells in cell culture as compared to fluorescence in situ hyberdisation (FISH) or quantitative polymerase chain reaction (qPCR). Both techniques are representing molecular high technology methods studying and amplifying both cell nuecleus ontent Deoxyribonucleic acid (DNA) and Ribonucleic acid (RNA) of myeloma cells. Furthermore FISH detects chromosomal aberration in both actively dividing cells and interphase nuclei. Therefore our approach utilises patients blood and bone marrow samples at time of diagnosis or relapse assessed by commercially available probes for FISH analysis on CD 138+ cells and reliably detects the prognostically significant genomic aberration, thus allowing clinicians to assess the biological risk of disease progression in patients with multiple myeloma (MM). The bone marrow studies will be performed as part of routine investigations for myeloma with extra 10 ml sample to conduct this research. The procedure takes approximately 30 min. This project will involve the validation of Fluorescence In-Situ Hybridisation (FISH) analysis of the 13q- and t(4:14) chromosomal abnormalities and development of rapid qPCR assays for the detection of these chromosomal abnormalities in bone marrow samples from denovo patients with MM. The aim of this project is to develop rapid qPCR assays for analysis of 13q- and t(4:14) chromosomal abnormalities and correlate the frequency of these abnormalities with treatment modalities and outcomes. The impact of these abnormalities in the context of high-dose therapy (HDT) or conventional therapy will help to identify those patients who would maximally benefit from HDT and those who would be refractory to HDT and might benefit from other therapeutic options. The Launceston General Hospital (LGH) is a tertiary referral centre of North and North West of Tasmania. We currently have at the LGH, 35-40 patients diagnosed with multiple myeloma with approximately 15-20 new cases every year and this observationall study will run for a total of 4 years ending in Dec 2011.