Study of aneuploidy in Multiple Myeloma

Postgraduate Thesis uoadl:2820019 434 Read counter

Unit:
Κατεύθυνση Εφαρμογές της Βιολογίας στην Ιατρική
Library of the School of Science
Deposit date:
2018-11-21
Year:
2018
Author:
Liakou Eleni
Supervisors info:
Τσιτσιλώνη Ουρανία, Αναπληρώτρια Καθηγήτρια του τμήματος Βιολογίας, ΕΚΠΑ
Original Title:
Μελέτη ανευπλοειδιών στο Πολλαπλό Μυέλωμα
Languages:
Greek
Translated title:
Study of aneuploidy in Multiple Myeloma
Summary:
Multiple myeloma (MM) is a malignany of terminally differentiated plasma cells, and typically patients are infiltrated with clonal plasma cells in bone marrow and secrete monoclonal protein (known as M protein) in the serum and/or urine although there is no secretion in about 3% of patients. Chromosomal translocation and aneuploidy are the pimary evants of MM. M protein is produced by the abnormal plasma cells and drives the clinical manifestations of disease such as hypocalcaemia, renal insufficiency, anemia, and/or bone disease with lytic lesions or pathological fractures, collectively known as CRAB features. Multiple myeloma is part of a range of disorders referred to as the monoclonal gammopathies, with the most common being monoclonal gammopathy of undetermined significance (MGUS). MGUS is asymptomatic and consistently precedes the development of multiple myeloma, with or without an identified intervening stage, referred to as smouldering multiple myeloma (SMM). Much progress has been made over the past decade in the understanding of disease biology and individualized treatment approaches. Several new classes of drugs, such as proteasome inhibitors and immunomodulatory drugs, have joined the traditional armamentarium (corticosteroids, alkylating agents and anthracyclines), along with high-dose therapy and autologous hematopoietic stem cell transplantation.
In this study simultaneous analysis for two colour immunophenotyping and DNA ploidy of plasma cells is described. 7AAD (7-aminoactinomycin D) dye which intercalates DNA, is used to stoichiometrically correspond to the fluorescence intensity of DNA with the number of chromosomes in a bone marrow sample, as well as, the establishment of a detectable threshold of hypodiploid plasma cells, which according to the literature is characterized of poor prognosis.
Thus, the immunophenotype of seven bone marrow samples have been tested in order to evaluate DNA ploidy corresponding to fluorescence of CD138-positive myeloma cells, in chromosome number. Required settings have been made on the flow cytometer in a view to quantifying fluorescence in FL3 channel, in which 7-AAD fluorescence is collected, in a linear scale. Under these settings, peripheral blood mononuclear cells and normal bone marrow, have been cultured with a combination of thymidine and colchicine to correspond the fluorescence intensity values of 46 and 92 chromosomes. In addition, the K562 cell line has been used as a third definition point of linearity as its chromosomes have been counted by karyotype. The intensity values are edited with the FORECAST function to produce ploidy results and compared with that of FISH and karyotype.
Main subject category:
Science
Keywords:
Multiple Myeloma, aneuploidy, bone marrow, chromosomal translocations
Index:
No
Number of index pages:
0
Contains images:
Yes
Number of references:
75
Number of pages:
113
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