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RISK FACTORS FOR THROMBOEMBOLIC COMPLICATIONS IN PATIENTS WITH
Ph-NEGATIVE MYELOPROLIFERATIVE DISEASES
Musashaykhova Sh.M., Musashaykhov Kh.T.
Andijan state medical institute
Abstract.
In this paper, we studied theая prognostic significance rof the rs1801133 polymorphic
locus of the MTHFR gene in the developmentof venous thromboembolic complications in
patients with Ph-negative MPD.
The results obtained indicate that there is a tendency to form a hypercoagulable syndrome and the
risk of thrombosis in patients with MPD with the presence of the unfavorable 677T allele of the
MTHFR gene in the genotype. The results of the study indirectly indicate the regulatory
"suppressive" effect of allelic variants 677T of the rs1801133 polymorphism of the MTHFR gene
on the production of the MTHFR enzyme, leading to hyperhomocysteinemia, which is an
important confirmation of the significance of genetic factors in the div's susceptibility to various
thromboembolic complications in MPD.
Key words:
Ph-negative myeloproliferative diseases, methylenetetrahydrofolate reductase,
hyperhomocysteinemia, thromboembolism.
Relevance
of
the
problem.
Chronic
Ph-negative
миелопролиферативные
myeloproliferativeоdiseases (MPD) are a group of pathogenetically identical hematological
diseases, includingих erythremia, essential thrombocythemia (ET), and ideopathic myelofibrosis
(IMF) [3, 6, 13]. These diseasesare caused by chronic leukemias with damage at the level of the
hematopoiesis progenitor cell with unlimited proliferation of this cell characteristic of the tumor,
the descendants of which are differentiated by all hematopoietic sprouts. (4, 7, 11). Classical
chronic MPDs are acquired sporadic disorders of hematopoiesis, but hereditary forms of
myeloproliferative diseases are also known-familial erythremia and thrombocythemia. Diagnosis
and treatment of chronic MPD is an urgent problem of modern hematology. Until recently, the
molecular defects and corresponding markers of IP, ET, and IMF were unknown. Therefore, the
arsenal of routine laboratory methods used to diagnose classical Ph-negative chronic MPD was
limited to the method of obtaining endogenous erythropoietin-independent colonies (EEC),
evaluating clonality, and determining the level of erythropoietin and thrombopoietin in the blood
of patients [1, 8, 12]. The recently increased interest in this disease is due to a significant increase
in the frequency of diagnosis of chronic MPD in patients of different ages, as well as a high risk
of developing thromboembolic complications in this category of patients, which are the main
cause of adverse outcomes and mortality (2, 10).
Methylenetetrahydrofolate reductase (MTHFR) isaurinary regulator of homocysteine metabolism
in the blood. Carriage of an unfavorable genotypic variant of polymorphism C677T (rs1801133)
of the MTHFR gene, which synthesizes a protein with reduced activity and increased thermal
stability, is associated with the development of hyperhomocysteinemiain plasma, which is an
independent risk factor for the development of various thromboembolic complications.
Hypercoagulable condition in the form ofvenous thrombosis is one of the most serious
complications in patients with Ph-negative MPD.
Despite intensive research, data on the role локуса C677of the C677t locus of the MTHFR gene
in the formation of individual predisposition to certain thrombotic complications among patients
with Ph-negative MPD remain very contradictory. This was the basis for conducting this study.
Patients with MPH have a high risk of thrombotic complications, disease progression with
transformation into secondary acute myeloid leukemia, and blast crisis [5, 9]. Due to the
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importance of molecular markers for both diagnosis and assessment of the risk of complications
and the risk of disease progression, this study will allow us to assess the role C677of the C 677t
polymorphism of the MTHFR gene in the development of thromboembolic complications in
patients with Ph-negative myeloproliferative diseases.
Goals and objectives of the study.
To assess the prognostic significance of polymorphic locus
rs1801133 of the MTHFR gene with the development of venous thromboembolic complications
in patients with Ph-negative MPD.
Materials and methods.
The study included DNA samples from 69 patients with MPD (39-
erythremia, 17-essential thrombocythemia, 13-subleukemic myelosis). Among these patients, 30
had various forms of thromboembolic complications. Genomic DNA extraction from peripheral
blood cells was performed using the Ampli PrimeRIBOT Prep kit. Detection of the rs1801133
locus of the MTHFR gene was performed by allele-specific PCR on a thermocycler Applied
Biosistems-2720 (USA), using kits of Litech LLC (Moscow), according to the manufacturer's
instructions.
Statistical processing of the results (case-control design) was performed using the OpenEpi
statistical software package (ver. 9. 3).
The results obtained and their discussion.
In the main and control groups, the level of observed heterozygosity did not exceed the
theoretically expected values, and the rs 1801133 locus1801133 of the MTHFR gene was in
accordance with the Hardy-Weinberg equilibrium (p>0.05).
In the studied groups of patients with MPD with and without thromboembolic complications, the
677C allele prevailed in frequencyС, occurring in 66.8% (41/60) and 75.4% (63/78), respectively.
In the subgroup of patients with thrombosis, there was a tendency to increase the frequency
of the 677T a allele associated with reduced MTHFR по срactivity in comparison with patients
without thrombosis (31.7% vs. 19.2%, respectively). The calculated relative chance of detecting
this allele in patients with thrombosis compared to the subgroup without thrombosis was OR=1.9
(χ
2
=2.8; p=0.09). The relative for the formation of hypercoagulation syndrome and the
development of thrombosis was RR= 1.6.
The frequency of occurrence of C/C, C/T, and T/T genotypes in the studied subgroup of patients
with and without thrombosis was 43.3%, 50.0%, and 6.7% versus 64.1%, 31.3%, and 2.6%,
respectively. The wild C/C genotype was associated with a protective effect on the div's
susceptibility to hypercoagulable syndrome (43.3% and 64.1%, respectively; χ
2
=3.0; p=0.09;
OR=0.4). The heterozygous S/T genotype was recorded significantly more often (a tendency to a
significant difference) in patients with thrombosis, than in the control group (50.0% vs. 31.3%,
respectively). According to the calculated odds ratio, the carrier of this genotype tended to
increase the riskа рof developing thrombosis by 2.0 times (χ
2
=1.9; p=0.2,; OR=2;), which
confirms the version about the involvement of this marker in the development of various
thrombotic complications in this pathology.
The homozygous T/T genotype was found only in 2 patients (6.7%) with thrombosis and in 1
without thrombosis (6.7% vs. 2.6%, with p>0.05). The odds ratio was OR=2.6, the relative risk
of thrombosis was RR=2.6.
The data obtained indicate that the presence of the rs 1801133 polymorphiclocus1801133 of the
MTHFR gene is an independent factor that increases the risk of thrombotic complications. The
occurrence of thrombotic complications is a criterion for the unfavorable development of chronic
MPD and can be consider as a factor in the negative prognosis of the disease. Along with the
leading factors that increase the risk of thrombosis (age over 60 years and a history of
cardiovascular risk factors), mutational status can also affect the course of MPD. Thus, the carrier
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of a mutation of the rs1801133 polymorphic locus of the MTHFR gene is associated with a
significant increase in the risk and frequency of thrombosis in this pathology.
Thus, the method developed by us for quantitative determination of the rs1801133 polymorphic
locus of the MTHFR gene by PCR highly specific, sensitive, and can complement morphological,
cytogenetic, and other studies. It allows a more adequate assessment of the risk of developing
thromboembolic complications, a deep study of the dynamics of minimal residual disease during
treatment with various drugs in patients with MPD, allowing not only to diagnose, but also to
quantify the effectiveness of specific therapy.
The data available in the literature on the role of these individual genetic markers are few and
contradictory. Only a detailed study of the population features of the above-mentioned genetic
mutations and an assessment of the correlation between the genotype and phenotype of the
disease can make it possible to choose the right strategy for early diagnosis and prognosis, as well
as the development of preventive measures for thromboembolic complications in patients with
MPD. The obtained data will improve the assessment of the clinical and prognostic significance
of the carriage of molecular genetic rearrangements in MPD and will contribute to updating
therapeutic approaches and algorithms, which will optimize the treatment and personalize the
tactics of therapy for this disease.
Conclusion.
The study showed, that molecular genetic analysis of mutations in the rs1801133
polymorphic locus of the MTHFR gene plays an exceptional role in assessing the risk of
thrombotic complications in classic Ph-negative MPD. It can be included in the disease prognosis
assessment scale as an independent factor. Additional studies are needed to clarify the role of
other molecular events in the formation of the phenotype of each individual nosology in the group
of Ph-negative MPD.ЗThe new data are of indisputable importance for the synthesis of targeted
drugs. Themolecular pathogenesis of chronic MPD is associated with somatic mutations of
regulatory genes. This observation serves as a strong argument in favor of the fact that when
further studying the molecular causes of occurrence, development, as well as clinical diversity,
risk assessment of thrombotic complications in MPD, special attention should be paid to the
significance of the polymorphic locus rs1801133 1801133 of the MTHFR gene.
Thus, the obtained data indicate that there is a tendency to form a hypercoagulable syndrome and
the risk of thrombosis in patients with MPD with the presence of the unfavorable 677T allele of
the MTHFR gene in the genotype. The results of the study indirectly indicate the regulatory
"suppressive" effect of allelic variants 677T of the rs1801133 polymorphism of the MTHFR gene
on the production of the MTHFR enzyme, leading to hyperhomocysteinemia, which is an
important confirmation of the significance of genetic factors in the susceptibility of the div to
various thromboembolic complications in hematological cancer patients.
A timely diagnosis and regular monitoring of treatment using clinical, morphological, cytogenetic
and molecular genetic research methods is a prerequisite for correct prediction of the course of the
disease and achieving maximum effectiveness of therapy. It seems appropriate to study the impact
of genetic rearrangements on the clinical course, possible potentiation of the risks of
thromboembolic complications, and overall prognosis of MPD.
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