SCIENCE AND INNOVATION IN THE
EDUCATION SYSTEM
International scientific-online conference
43
TO STUDY THE RELATIONSHIP OF TNFα (G-308A) GENE
POLYMORPHISM IN THE DEVELOPMENT OF APLASTIC ANEMIA
Akhmedova Zukhra Bakhtiyarovna
basic doctoral student
RSSPMCH, number: +99894 726 87 88.
Matkarimova Dilfuza Saburova
Professor of the Department of Hematology, Transfusiology and Laboratory
Science of TMA, number: +99897 412 91 18.
Boboev Kodirjon Tukhtabaevich
Head of the Department of Molecular Medicine and Cellular Technologies of the
RSSPMCH, number: +99890 319 39 57.
Republican specialized scientific - practical medical center of Hematology
Tashkent medical academy
https://doi.org/10.5281/zenodo.14868537
Summary
Purpose of the study.
To analyze the functional significance of
polymorphism of the proinflammatory cytokine TNFa (G-308A) gene in the
development of aplastic anemia.
Methods.
Detection of polymorphic loci of the TNFa gene (rs1800629) in
86 patients with AA and 98 healthy ones using polymerase chain reaction in
standard mode with visualization of electrophoresis products on a
programmable thermal cycler "Rotor Gene Q" (Quagen, Germany).
In groups of patients with AA and healthy, a molecular genetic
analysis was performed with DNA isolation from peripheral blood using a set of
reagents "AmpliPrime RIBOT-prep" (Russia) and detection of TNFa genetic
polymorphism (rs1800629) using test systems "Litech, NPF LLC" (Russia). The
amplification process was reproduced on the GeneAmp PCR-system 2720
thermal cycler (Applied Biosystems, USA). The amplified products were
subjected to electrophoresis in 2% agarose gel to study band patterns using
ethidium bromide. Statistical processing of the obtained results was carried out
using the OpenEpi – 2009 software package (Version 2.3).
Conclusions.
Genetic polymorphism TNFα (rs1800629) is not associated
with the risk of developing aplastic anemia.
Key words:
Tumor necrosis factor-α TNFa (rs1800629), single nucleotide
polymorphism (SNP), autoimmune disease, proinflammatory cytokine.
Introduction.
Tumor necrosis factor alpha (TNF-α) is a pro-inflammatory
cytokine produced by various immune cells, including antigen-stimulated T cells,
lymphocytes and NK cells [2,4,5]. Its effect is to limit concomitant damage to
SCIENCE AND INNOVATION IN THE
EDUCATION SYSTEM
International scientific-online conference
44
host cells and tissues during an inflammatory reaction and to maintain a balance
between inflammatory and anti-inflammatory reactions [1,9,11].
TNF-α (tumor necrosis factor-α) is important for the normal functioning of
the div, but is also involved in some disease mechanisms, including sepsis,
diabetes mellitus, and cancer [8,10].
Recent studies have shown that defective functioning of regulatory T cells
leads to increased production of interferon gamma (IFN-γ) and tissue necrosis
factor (TNF-α), causing damage to stem cells, which leads to bone marrow
aplasia [3,6,7,12].
To determine the distribution of SNP loci of the TNFa polymorphic gene (G-
308A) and its connection with the formation of AA, we performed a molecular
and genetic analysis of this polymorphism.
Results.
The assumptions made were also proved by the results of
mathematical analyses of differences between polymorphic loci of the TNFa (G-
308A) gene in the studied groups.
Statistically insignificant differences were found between the frequencies of
alleles and genotypes of the TNFa (G-308A) polymorphism in the main and
control groups, amounting to less than one for the mutant A allele (5.2% vs.
5.6%; χ2<3.84; P=0.9; RR=1.0; CI: 0.45-2.22; OR=0.9; CI: 0.38 - 2.3), with a risk of
developing the disease equal to 1.1 (OR) – for the main genotype (89.5% vs.
88.8%; -2<3.84; P=0.9; RR=1.0; CI: 0.37-2.75; OR=1.1; CI: 0.43-2.75) and less
than one for the G/A heterozygote (10.5% vs. 11.2%; χ2<3.84; P=0.9; RR=0.9;
DI: 0.34-2.54; OR=0.9; DI: 0.36-2.35)
According to the polymorphic TNFa gene (G-308A), no statistically
significant differences were found in the frequencies of allelic and genotypic
variants between groups of patients with mild AA and healthy patients.The
proof of this was the absence of significant differences among carriers of the
mutant A allele (3.1% vs. 5.6%; χ2=0.3; P=0.6; RR=1.0; DI: 0.69-1.4; OR=0.5; DI:
0.07-4.22), the main G|G homozygote (93.8% vs. 88.8%; χ2=0.4; P=0.6; RR=1.1;
DI: 0.02-46.7; OR=1.9; DI: 0.24-15.3) and the heterozygote G/A (6.3% vs. 11.2%;
χ2=0.4; P=0.6; RR=0.6; DI: 00.01- 4.6; OR=0.5; DI: 0.07-4.25)
A similar analysis between groups of patients with severe AA and healthy
controls confirmed the absence of statistically significant differences among
carriers of polymorphic TNFa (G-308A) gene loci: attenuated A allele (5.4% vs.
5.6%; χ2<3.84; P=0.98; RR=1.0; CI: 0.51-1.94; OR=0.5; DI:0.33-2.87), the main
homozygote G/G (89.1% vs. 88.8%; χ2<3.84; P=0.95; RR=1.0; DI: 0.22-4.53;
OR=1.0; CI: 0.34-3.18) and G/A heterozygotes (6.3% vs. 11.2%; χ2<3.84; P=0.95;
SCIENCE AND INNOVATION IN THE
EDUCATION SYSTEM
International scientific-online conference
45
RR=1.0; CI: 0.21-4.37; OR=1.0; DI: 0.31-2.96).This means that the polymorphic
loci of the TNFa gene (G-308A) do not have an independent role in the formation
of severe AA.
In the structure of the TNFa polymorphic gene (G-308A), a two-way
comparative analysis between groups of patients with superheavy AA and
healthy ones allowed us to determine the absence of statistically significant
differences in the frequencies of allelic and genotypic variants.Thus, among
patients, the frequency of the mutant allele A (6.3% vs. 5.6%; χ2<3.84; P=0.9;
RR=1.0; DI: 0.58-1.75; OR=1.1; CI: 0.3-4.18) and heterozygous genotype G/A
(12.5% vs. 11.2%; χ2<3.84; P=0.9; RR=1.1; CI: 0.14-9.14; OR=1.1; CI:0.29-4.41)
turned out to be slightly more than one, and the difference between the studied
groups in the frequency of the main homozygote G/G did not even reach one
(87.5% vs. 88.8%; -2<3.84; P=0.9; RR=1.0; CI: 0.12-8.1; OR=1.9; DI: 0.23-3.46)
and heterozygous.The results obtained serve as evidence of the absence of an
independent relationship between polymorphic loci of the TNFa gene (G-308A)
and an increased risk of formation of the superheavy form of AA.
Analyzing the significance of differences in the frequencies of polymorphic
loci of the TNFa gene (G-308A) in a group of patients with mild AA compared
with severe and superheavy forms of AA, there were significant differences in
the carriage of the mutant A allele (3.1% vs. 5.4%; χ2=0.3; P=0.6; RR=1.0; DI:
0.47 - 2.03; OR=0.6; DI: 0.06-4.86 and 3.1% vs. 6.3%; χ2=0.4; P=0.6; RR=1.0; DI:
0.3-3.11; OR=0.5; DI: 0.05-4.66), the main genotype is G/G (93.8% vs. 89.1%;
χ2=0.3; P=0.6; RR=1.1; DI: 0.03-38.79; OR=1.8; DI: 0.2-16.49 and 93.8% vs.
87.5%; χ2=0.4; P=0.6; RR=1.1; DI: 0.04-32.48; OR=2.1; DI: 0.21-21.68) and
heterozygous G/A (6.3% vs. 10.9%; χ2=0.3; P=0.6; RR=0.6; DI: 0.02- 21.2;
OR=0.5; DI: 0.06-4.93 and 6.3% vs. 12.5%; χ2=0.4; P=0.6; RR=0.5; DI: 0.02-
15.16; OR=0.5; DI: 0.05-4.72) was not detected.
Along with this, comparing the degree of differences in the frequencies of
polymorphic loci of the TNFa gene (G-308A) between severe and superheavy
forms of AA, there were significant differences in the carriage of the mutant A
allele (5.4% vs. 6.3%; χ2<3.84; P=0.9; RR=1.0; CI: 0.16-6.08; OR=0.9; CI: 0.2-
3.77), the main genotype G/G (89.1% vs. 87.5%; -2<3.84; P=0.9; RR=1.0; CI:
0.34-3.09; OR=1.2; DI: 0.26-5.38) and heterozygous G/A (10.9% vs. 12.5%;
χ2<3.84; P=0.9; RR=0.9; DI: 0.29- 2.63; OR=0.9; DI: 0.19-3.92) has also not been
established.
SCIENCE AND INNOVATION IN THE
EDUCATION SYSTEM
International scientific-online conference
46
Conclusion:
The results obtained serve as evidence of the absence of an
independent relationship between polymorphic loci of the TNFa gene (G-308A)
and an increased risk of AA formation and its severe course in Uzbekistan.
References:
1.
Alkhuriji A. F. et al. Association of IL-1β, IL-6, TNF-α, and TGFβ1 gene
polymorphisms with recurrent spontaneous abortion in polycystic ovary
syndrome //Disease Markers. – 2020. – Т. 2020. – №. 1. – С. 6076274.
2.
Barnes D. W. H., Mole R. H. Aplastic anaemia in sublethally irradiated mice
given allogeneic lymph node cells. – Medical Research Council, Harwell, Eng.,
1967.
3.
Furlong E., Carter T. Aplastic anaemia: Current concepts in diagnosis and
management //Journal of paediatrics and child health. – 2020. – Т. 56. – №. 7. –
С. 1023-1028.
4.
Hinterberger W. et al. Results of transplanting bone marrow from
genetically identical twins into patients with aplastic anemia //Annals of
internal medicine. – 1997. – Т. 126. – №. 2. – С. 116-122.
5.
Jang H. G. et al. Polymorphisms in tumor necrosis factor-alpha (− 863C>
A,− 857C> T and+ 488G> A) are associated with idiopathic recurrent pregnancy
loss in Korean women //Human Immunology. – 2016. – Т. 77. – №. 6. – С. 506-
511.
6.
Kali, Arunava. "TNFerade, an innovative cancer immunotherapeutic."
Indian Journal of Pharmacology 47.5 (2015): 479-483.
7.
Luzzatto L, Risitano AM. Advances in understanding the pathogenesis of
acquired aplastic anaemia. Br J Haematol. 2018; 182:758–76. doi:
10.1111/bjh.15443.
8.
Medinger M. et al. Pathogenesis of acquired aplastic anemia and the role of
the bone marrow microenvironment //Frontiers in oncology. – 2018. – Т. 8. – С.
587.
9.
Neumann C., Scheffold A., Rutz S. Functions and regulation of T cell-
derived interleukin-10 //Seminars in immunology. – Academic Press, 2019. – Т.
44. – С. 101344.
10.
Qiao, Yong-chao, et al. "The change of serum tumor necrosis factor alpha in
patients with type 1 diabetes mellitus: A systematic review and meta-analysis."
PloS one 12.4 (2017): e0176157.
11.
Wang X. A. et al. Mesenchymal stem cells in acquired aplastic Anemia: the
Spectrum from Basic to Clinical Utility //International Journal of Molecular
Sciences. – 2023. – Т. 24. – №. 5. – С. 4464.
SCIENCE AND INNOVATION IN THE
EDUCATION SYSTEM
International scientific-online conference
47
12.
Zhang Y, Cui X, Ning L, Wei D. The effects of tumor necrosis factor-α (TNF-
α) rs1800629 and rs361525 polymorphisms on sepsis risk. Oncotarget.
2017;8(67):111456–69.
