Download AS A PROGNOSTIC MARKER IN CHRONIC MYELOID LEUKEMIA

RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES, BANGALORE,
KARNATAKA
PROFORMA FOR REGISTRATION OF SUBJECTS FOR DISSERTATION
1. NAME OF THE CANDIDATE:
ADDRESS:
Dr. Christine Mary Jane
Department of Medicine
St. John’s Medical College Hospital
John Nagar, Sarjapur Road
Koramangala
Bangalore – 560034
Tel. No. 080-22065834
2. NAME OF THE INSTITUTION:
St. John’s Medical College
Bangalore
3. COURSE OF STUDY AND SUBJECT:
MD, General Medicine
4. DATE OF ADMISSION TO THE COURSE:
01-06-2013
5. TITLE:
STUDY OF CNVs COPY NUMBER
VARIANTS) AS A PROGNOSTIC
MARKER IN CHRONIC MYELOID
LEUKEMIA.
STUDY OF CNVs (copy number variants) AS A PROGNOSTIC MARKER IN
CHRONIC MYELOID LEUKEMIA
DR CHRISTINE MARY JANE
GUIDE: DR CECIL ROSS MD
PROFESSOR.
6. Introduction
 CML is a clonal, haematologic malignant disease characterised by the
acquisition of the BCR/ABL fusion gene generated by a cytogenetically
balanced philadelphia chromosome translocation.
 The product of the gene, the bcr/abl protein is a constitutively active
protein tyrosine kinase, which plays a central role in the pathogenesis of
CML(1).
 There is variability in drug sensitivity, resistance and response to
treatment in the three disease phenotypes of CML- chronic phase,
accelerated phase and blast phase.
6.1Study agent/procedure
 The development of drug resistance is multi factorial.
 It can include mechanisms such as BCR/ABL gene amplification,
development of multidrug resistance or mutation of the BCR/ABL kinase
domain.
 But not all treatment failures can be accounted for by these changes.
6.2 CNV (COPY NUMBER VARIATION)
 CNV (Copy Number Variation) can be defined as a DNA segment that is 1
kbp or larger and present at variable copy number in comparison with a
reference genome.
 Deletions, insertions, duplications and complex multi-site variants are
included under CNVs.
 A CNV can be simple in structure, such as tandem duplication, or may
involve complex gains or losses of homologous sequences at multiple
sites in the genome.
 CNVs influence gene expression, phenotypic variation and adaptation by
disrupting and altering gene and can cause diseases (2).
6.3 Objective:
To assess prognostic significance of known CNVs in CML patients on Tyrosine
kinase inhibitor therapy (TKI) therapy.
Inclusion criteria :
BCR-ABL positive CML patients in chronic/
accelerated phase/ blast crisis who have been on TKI therapy for a minimum
of 3 months
Exclusion criteria
 Clinical evidence of heart failure.
 Concurrent serious hepatic disorder or renal disorder.
 On irregular treatment.
7.STUDY DESIGNA cross sectional study.
All patients with CML on TKI’s (tyrosine kinase inhibitors) for at least 3
months on follow up in St. John’s Medical college
 Duration of study:
One year from october 2013 to october 2014.
 Sample size: 30
 Sampling technique: Convenience sampling.
7.1 Criteria for Cytogenetic, Hematologic & Molecular Response :
 This is based on clinical analysis, cytogenetic study and hybridization
techniques.
7.2MATERIALS AND METHODS :
 Patients who have been on TKI for a minimum of 3 months will be
assessed by RQ-Polymerase chain reaction for determining CNVs.
 Comparison will be made between CNV in patients among the following
groups:
1. With complete haematological response (CHR) and depth of molecular
remission.
2. Without CHR
3. Blast crises and accelerated phase
Statistical analysis:
 The relative copy number of target sequences in DNA samples can be
determined using a comparative CT (Δ CT) method. This method
calculates a relative value both to the normal control DNA (the
callibrator) and to the endogenous control reference sequence.
7.3 Data analysis procedure:
Descriptive measure
 The change in a particular gene is a numerical variable. Drug response is
a dichotomous variable- Yes/No.
 Hence a student T test will be done for each gene studied.
FUNDING: Intramural funds.
8.References:
1.Markovic VD, Bouman D, Bayani J, Al-Maghrabi J, Kamel-Reid S,Squire
JA.;Lack of BCR/ABL reciprocal fusion in variant Philadelphia chromosome
translocations: a use of double fusion signalFISH and spectral karyotyping,
Leukemia 14(2000) 1157–1160.
2. Redon R, Ishikawa S, Fitch KR, Feuk L, Perry GH, Andrews TD, Fiegler
H,Shapero MH, Carson AR, Chen W, Cho EK, Dallaire S, Freeman JL, González
JR,Gratacòs M, Huang J, Kalaitzopoulos D, Komura D, MacDonald JR, Marshall
CR, Mei R, Montgomery L, Nishimura K, Okamura K, Shen F, Somerville MJ,
Tchinda J,Valsesia A, Woodwark C, Yang F, Zhang J, Zerjal T, Zhang J, Armengol
L, Conrad DF, Estivill X, Tyler-Smith C, Carter NP, Aburatani H, Lee C, Jones KW,
Scherer SW, Hurles ME. Global variation in copy number in the human
genome. Nature. 2006 Nov 23;444(7118):444-54.
3. Braude I, Vukovic B, Prasad M, Marrano P, Turley S, Barber D, Zielenska M,
Squire JA. Large scale copy number variation (CNV) at 14q12 is associated with
the presence of genomic abnormalities in neoplasia. BMC Genomics. 2006 Jun
6;7:138. PubMed PMID: 16756668; PubMed Central PMCID: PMC1550726.
4. Branford S, Hughes T. Diagnosis and monitoring of chronic myeloid leukemia
by qualitative and quantitative RT-PCR. Methods Mol Med. 2006;125:69-92.
5. Kolomietz E, Al-Maghrabi J, Brennan S, Karaskova J, Minkin S, Lipton J, Squire
JA. Primary chromosomal rearrangements of leukemia are frequently
accompanied by extensive submicroscopic deletions and may lead to altered
prognosis. Blood. 2001 Jun 1;97(11):3581-8.
6.Kolomietz E, Marrano P, Yee K, Thai, Braude I, Kolomietz A, K Chun, S
Minkin1, Kamel-Reid S,Minden1 M and Squire JA; Quantitative PCR identifies a
minimal deleted region of 120 kb extending from the Philadelphia
chromosome ABL translocation breakpoint in chronic myeloid leukemia with
poor outcome. Leukemia. 2003 Jul;17(7):1313-23.
7.Guan M, Liu L, Zhao X,Wu Q,Yu B, Shao Y, Yang H, Fu X,Wan J, Zhang W; Copy
Number Variations of EphA3 Are Associated With Multiple Types of
Hematologic Malignancies, Clinical Lymphoma, Myeloma & Leukemia.
Clin Lymphoma Myeloma Leuk. 2011 Feb;11(1):50-3.
9. SIGNATURE OF THE CANDIDATE
10. REMARKS OF THE GUIDE
11. NAME AND DESIGNATION OF
11.1 GUIDE:
DR. CECIL ROSS
PROFFESSOR OF MEDICINE
11.2 SIGNATURE:
11.3 HEAD OF DEPARTMENT:
DR.G.D.RAVINDRAN
PROFESSOR & HEAD
11.4. SIGNATURE:
11.5. REMARKS OF CHAIRMAN AND DEAN:
11.6. SIGNATURE:
DR.