engleski

Notch pathway genes' analysis in multiple myeloma remission state

Multiple myeloma (MM), a second most common haematological malignancy, is a B cells disorder characterized by rapid clonal plasma cells' proliferation and monoclonal gammopathy, leading to severe bone disruptions, hypercalcemia, and abnormal function of the immune system. The origin and the disease developing mechanism are unknown and there is no cure for MM. Existing therapies are sometimes able to halt the disease, but, in most cases, absence of the disease or the remission state is only temporary and MM relapses. Notch signalling pathway can be deregulated in the state of the disease. This signalling pathway consists of transmembrane receptors NOTCH which activate by binding to the neighbouring cells' transmembrane ligands DELTA or JAGGED. Ligand binding causes cleavage of the NOTCH intracellular domain, which acts as a part of transcription complex in the nucleus. MM cells either have overexpression of NOTCH receptors or other mechanisms, which lead to the pathway activation. The aim of the thesis was to see if NOTCH pathway deregulation could be detected in cells obtained from patients with multiple myeloma in remission. Thus, we analysed Notch pathway genes’ expression in peripheral blood samples of MM patients in remission. Peripheral blood samples and a fraction of B cells were analysed for the expression of NOTCH receptors, JAGGED and DELTA ligands, downstream target HES1, as well as differentiation factors from IKAROS family. The results showed no uniform pattern which could connect the NOTCH pathway deregulation to the relapse. For the analysis of small cell number samples from selected B cells we optimized a multiplex, semi nested PCR method. This method is valuable as it enables inspection of multiple different genes simultaneously, on a very small cell number, with increased specificity and sensitivity.

multiple myeloma, B cell, NOTCH signalling pathway, remission, multiplex semi nested PCR

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