Molecular Genetic Characteristic of Pulmonary Tuberculosis Associated with ABCB1 Gene Expression of Multidrugresistance Protein P-gp

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Abstract

Background. Tuberculous inflammation is mediated by a complex molecular signaling pathway, the analysis of which makes it possible to identify promising biomarkers and targets for the development of new diagnostic, prognostic and pharmacological approaches in order to improve the effectiveness of anti-tuberculosis chemotherapy. Determining the relationship between key inflammatory cytokines, the multidrug-resistant protein P-gp and the activity of specific inflammation in the surgical material of patients with pulmonary tuberculosis may prove to be a novel tool in the development of pathogenetic therapy and personalized medicine.

Aims — to characterize molecular and genetic profiles of tuberculomas and identify genes that correlate with the expression of the ABCB1 gene of the P-gp protein in the surgical material of patients with pulmonary tuberculosis. Research objectives: 1) to obtain molecular and genetic characteristics of tuberculosis by real-time PCR and compare it with the activity of tuberculous inflammation; 2) to carry out a correlation analysis between the expression of the ABCB1 gene and key cytokines of the tuberculosis process: IL-6, IL-10, IFN-γ, TGF-β, TNF-α, IL-1β.

Methods. A prospective cohort study was conducted on the basis of the FSBI CTRI. The object of the study was the surgical material of 35 patients diagnosed with multiple pulmonary tuberculomas. Histological examination methods were used for the morphological assessment of the surgical material. A real-time quantitative PCR method was used to analyze gene expression. Statistical processing was performed using the GraphPad Prism Version 7.04 software package (GraphPad Software, USA). The data is presented as a median with an interquartile range. The nonparametric Mann–Whitney U-test was used to compare the two groups. All p-values were two-sided and p < 0.05 was considered statistically significant. The correlation between the variables was estimated using the Spearman correlation coefficient. The correlation analysis was carried out in the Microsoft Office Excel 2010 Software.

Results. The study revealed that the highest level of expression of ABCB1 gene of the P-gp protein is observed in tuberculomas with high activity of tuberculous inflammation, and its expression is correlated with the expression of the IL6 gene (p < 0.001) and the expression of the IL10 gene (p < 0.01). Tuberculomas of this group are also characterized by higher expression of the TGFB1, TNF and IL1B genes, compared with the group of moderate activity of specific inflammation.

Conclusions. The data obtained indicate that in addition to pro-/anti-inflammatory cytokines, the P-gp protein plays an important role in the pathogenesis of tuberculous inflammation, especially with its high activity. Further clarification of the P-gp role in tuberculous inflammation may be an important step for the development of new approaches to treat tuberculosis using methods of HDT and personalized medicine.

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About the authors

Atadzhan E. Ergeshov

Central Tuberculosis Research Institute; Russian University of Medicine

Email: cniit@ctri.ru
ORCID iD: 0000-0002-2494-9275
SPIN-code: 8372-1666

д.м.н., профессор, член-корреспондент РАН

Россия, Moscow; Moscow

Maria V. Erokhina

Central Tuberculosis Research Institute; Lomonosov Moscow State University

Author for correspondence.
Email: masha.erokhina@gmail.com
ORCID iD: 0000-0002-7256-4679
SPIN-code: 8033-0860

PhD in Biology, Associate Professor

Россия, Moscow; Moscow

Ekaterina N. Pavlova

Lomonosov Moscow State University

Email: guchia@gmail.com
ORCID iD: 0000-0001-9498-1142
SPIN-code: 8311-7260

PhD in Biology

Россия, Moscow

Larisa N. Lepekha

Central Tuberculosis Research Institute

Email: lep3@yandex.ru
ORCID iD: 0000-0002-6894-2411
SPIN-code: 6228-8382

PhD in Biology, Professor

Россия, Moscow

Ruslan V. Tarasov

Central Tuberculosis Research Institute

Email: etavnai@yandex.ru
SPIN-code: 4245-1560

MD, PhD

Россия, Moscow

Ekaterina K. Tarasova

Central Tuberculosis Research Institute

Email: shalioto6@gmail.com
ORCID iD: 0000-0003-0438-7233
SPIN-code: 5661-8640

Master of Biology

Россия, Moscow

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2. Fig. 1. Tuberculomas with signs of moderate activity of the inflammatory process: A — general appearance; B, C — foci of seeding in the form of granulomas or caseous necrosis with signs of organization. Stained with hematoxylin and eosin. Scale bar — 100 μm Note. CN — caseous necrosis; GS — granulation layer; FS — fibrous layer; EC — epithelioid cell granuloma.

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3. Fig. 2. Pulmonary tuberculomas with signs of high inflammatory activity: A — caseous necrosis with signs of melting, surrounded by a wide layer of granulation tissue containing cellular elements of inflammation and epithelioid cell granulomas; B — a focus of seeding in the form of epithelioid cell granulomas with and without necrosis merging with each other; C — a focus of caseous necrosis surrounded by granulation tissue without pronounced signs of organization. Hematoxylin and eosin staining. Scale bar — 100 μm Note. CN — caseous necrosis; GL — granulation layer; FL — fibrous layer; NP — area of ​​nonspecific pneumonia; EC — epithelioid cell granuloma.

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4. Fig. 3. Histograms demonstrating high expression levels of ABCB1, IL6, IL1B, TGFB1 genes in tuberculomas with high activity of specific inflammation

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5. Fig. 4. Matrix of significant Spearman correlation coefficients (r) between relative levels of gene expression in the lungs of patients diagnosed with multiple tuberculomas

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