Characteristics of the functional state of peripheral blood neutrophils in patients with luminal breast cancer

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Abstract

BACKGROUND: Neutrophils are essential in tumor growth, and their functional state can serve as a prognostic biomarker. However, the functional characteristics of peripheral blood neutrophils, such as chemotaxis and predisposition to NETosis, in female patients with luminal breast cancer have not been sufficiently explored. Studying these parameters may provide new insights into the mechanisms of disease progression and response to therapy.

AIM: This work aimed to analyze the chemotactic activity of neutrophils and predisposition to NETosis in blood samples of female patients with locally advanced luminal breast cancer undergoing treatment (neoadjuvant chemotherapy) at the Loginov Moscow Clinical Scientific Center.

METHODS: The study was conducted on blood samples from six patients with stage 3 luminal B, HER2-negative breast cancer before and 2 months after the start of antitumor therapy. Blood samples from healthy adult volunteers were used as controls. The work was performed using fluorescence microscopy methods for neutrophil chemotaxis with the growth of blood clots and the number of extracellular DNA traps of neutrophils by reaction with Hoechst 33342 and antibodies against myeloperoxidase and neutrophil elastase in smears of blood plasma rich in leukocytes.

RESULTS: Before the start of neoadjuvant therapy, the level of NETosis is significantly increased (30% ± 14% versus 4.6% ± 3.4% in healthy donors), whereas most female patients undergoing therapy experience its reduction (17% ± 17%). The speed of neutrophil movement is increased in some female patients (0.17 ± 0.06 versus 0.113 ± 0.009 μm/s in healthy donors) and goes down during therapy (0.10 ± 0.03 μm/s). At the same time, the number of neutrophils associated with blood clots decreases during therapy (25 ± 18 versus 61 ± 23) even in patients with neutrophilia.

CONCLUSION: It has been demonstrated for the first time that in female patients with luminal B, HER2-negative breast cancer, the neutrophil chemotaxis speed deviates from the standard; at the same time, their adhesion is reduced, and peripheral blood neutrophils are significantly more predisposed to NETosis than in healthy donors.

About the authors

Julia Jessica D. Korobkina

Center for Theoretical Problems of Physico-Chemical Pharmacology RAS

Email: juliajessika@gmail.com
ORCID iD: 0000-0002-2762-5460
SPIN-code: 6630-3657
Russian Federation, Moscow

Ekaterina-Iva A. Adamanskaya

Center for Theoretical Problems of Physico-Chemical Pharmacology RAS; Dmitry Rogachev National Research Center of Pediatric Hematology, Oncology, and Immunology

Email: ka.09@mail.ru
ORCID iD: 0009-0000-4828-4063
SPIN-code: 9633-1147
Russian Federation, Moscow; Moscow

Natalya I. Polshina

Loginov Moscow Clinical Scientific Center

Email: npolshina@yandex.ru
ORCID iD: 0000-0001-5417-0425

MD

Russian Federation, Moscow

Sofia V. Galkina

Center for Theoretical Problems of Physico-Chemical Pharmacology RAS; Dmitry Rogachev National Research Center of Pediatric Hematology, Oncology, and Immunology

Email: s_v_galkina@rambler.ru
ORCID iD: 0009-0006-6321-4489
Russian Federation, Moscow; Moscow

Timur I. Kadyrov

Center for Theoretical Problems of Physico-Chemical Pharmacology RAS

Email: kadyrov.ti17@physics.msu.ru
ORCID iD: 0009-0005-0130-1758
Russian Federation, Moscow

Nikolay P. Gorbunov

Institute for Experimental Medicine

Email: niko_laygo@mail.ru
ORCID iD: 0000-0003-4636-0565
SPIN-code: 6289-7281
Russian Federation, Saint Petersburg

Alexey V. Sokolov

Institute for Experimental Medicine

Email: biochemsokolov@gmail.com
ORCID iD: 0000-0001-9033-0537
SPIN-code: 7427-7395

Dr. Sci. (Biology)

Russian Federation, Saint Petersburg

Lyudmila G. Zhukova

Loginov Moscow Clinical Scientific Center

Email: zhukova.lyudmila008@mail.ru
ORCID iD: 0000-0003-4848-6938
SPIN-code: 2177-6476

MD, Dr. Sci. (Medicine)

Russian Federation, Moscow

Anastasia N. Sveshnikova

Center for Theoretical Problems of Physico-Chemical Pharmacology RAS; Dmitry Rogachev National Research Center of Pediatric Hematology, Oncology, and Immunology

Author for correspondence.
Email: ASve6nikova@yandex.ru
ORCID iD: 0000-0003-4720-7319
SPIN-code: 7893-4627

Dr. Sci. (Physics and Mathematics)

Russian Federation, Moscow; Moscow

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5. Fig. 1. Observation of NETosis in blood plasma smears. а, means a proportion of NETotic neutrophils. The Mann–Whitney criterion was used for comparison with healthy donors: **p < 0.01, ***p < 0.001. There is no statistically significant difference between the first and second points (according to the Wilcoxon signed-rank test when comparing without ЗCA, p = 0.06). The ratio of NETosis at the first (b) and second (c) study points. ЗД means a group of healthy donors (n = 8). ЗСА, МЮИ, МЕГ, МАА, КЕА, МСЕ are patient codes.

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6. Fig. 2. Experimental model of thromboinflammation: а, means the average neutrophil movement speed in the experiment; b, means the number of neutrophils adhering to the substrate during the experiment. For comparison purposes, the non-parametric Mann–Whitney test was used: *p < 0.05.

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