Monitoring of breast cancer progression via aptamer-based detection of circulating tumor cells in clinical blood samples

Introduction: Breast cancer (BC) diagnostics lack noninvasive methods and procedures for screening and monitoring disease dynamics. Admitted CellSearch® is used for fluid biopsy and capture of circulating tumor cells of only epithelial origin. Here we describe an RNA aptamer (MDA231) for detecting BC cells in clinical samples, including blood. The MDA231 aptamer was originally selected against triple-negative breast cancer cell line MDA-MB-231 using cell-SELEX.Methods: The aptamer structure in solution was predicted using mFold program and molecular dynamic simulations. The affinity and specificity of the evolved aptamers were evaluated by flow cytometry and laser scanning microscopy on clinical tissues from breast cancer patients. CTCs were isolated form the patients’ blood using the developed method of aptamer-based magnetic separation. Breast cancer origin of CTCs was confirmed by cytological, RT-qPCR and Immunocytochemical analyses.Results: MDA231 can specifically recognize breast cancer cells in surgically resected tissues from patients with different molecular subtypes: triple-negative, Luminal A, and Luminal B, but not in benign tumors, lung cancer, glial tumor and healthy epithelial from lungs and breast. This RNA aptamer can identify cancer cells in complex cellular environments, including tumor biopsies (e.g., tumor tissues vs. margins) and clinical blood samples (e.g., circulating tumor cells). Breast cancer origin of the aptamer-based magnetically separated CTCs has been proved by immunocytochemistry and mammaglobin mRNA expression.Discussion: We suggest a simple, minimally-invasive breast cancer diagnostic method based on non-epithelial MDA231 aptamer-specific magnetic isolation of circulating tumor cells. Isolated cells are intact and can be utilized for molecular diagnostics purposes

Параллельная пандемия: психологическая помощь в медицинской сфере

Analysis of current studies in Russia and other countries allows to make a conclusion about the growing pace, developing so-called parallel pandemic, which is formed at the level of psychological health. This situation reveals the urgency to analyze the situation of the COVID-19 pandemic from both clinical psychology and general medical practice points of view. This article attempts to highlight the potentially possible, negative consequences of the pandemic, which affect the psychological state of different categories of the population. In particular the special attention is paid to the difficulties faced by health-care staff at the core of the fight against COVID-19. A comparative analysis of the current situation allowed the authors of this article to draw parallels between the state in which most of the population is now living and the state of the learned helplessness. A scheme for the medical and psycho-logical assistance in the post-pandemic period has been proposed

Locus 9p21.3 – Genetic Predictor of Coronary Atherosclerosis Severity in Siberians

The purpose of this study was to investigate association between 9p21.3 locus single nucleotide polymorphisms (SNP) and severity of coronary atheromatous burden. A total of 255 Caucasians (211 male, 44 female) with myocardial infarction younger 65 years old (mean 52,56±7,98 years) were enrolled. Genome DNA was extracted from venous blood using the phenol-chloroform extraction method. Two SNPs rs10757278 and rs1333049 (locus 9p21.3) were tested using real-time polymerase chain reaction (PCR) according to protocol (probes TaqMan, Applied Biosystems, 7900HT). The coronary angiograms were reviewed by independent angiographers who were blinded to the results of the genotype (Philips Allura Xper FD10). The total number of lesions, Gensini scores and SYNTAX score were derived. For the first time in males of Siberian population, genotype CC rs1333049 and genotype GG rs10757278 demonstrated a direct strong association with severity of coronary atheromatous burdenЦель исследования – изучить взаимосвязь локуса 9р21.3 с тяжестью коронарного атеросклероза в сибирской популяции. В исследование включены 255 больных (211 мужчин, 44 женщины) с инфарктом миокарда (ИМ) европеоидной расы в возрасте ≤ 65 лет (52,56±7,98 лет). Геномную ДНК выделяли из венозной крови фенол-хлороформным методом. Полиморфизм генов тестировали с помощью полимеразной цепной реакции (ПЦР) в режиме реального времени (зонды TaqMan, Applied Biosystems 7900HT). В исследовании изучались два однонуклеотидных полиморфизма (ОНП), расположенные в локусе 9p21.3 – rs10757278 и rs1333049. Оценка степени тяжести поражения коронарного русла производилась в заслепленном режиме в ходе стандартной полипроекционной коронарографии (Philips Allura Xper FD10), учитывались количество пораженных сегментов, интегральные индексы Gensini и SYNTAX. Впервые в сибирской популяции у мужчин доказана взаимосвязь генотипов GG rs10757278 и СС rs1333049 с тяжестью коронарного атеросклероз

Locus 9p21.3 – Genetic Predictor of Coronary Atherosclerosis Severity in Siberians

The purpose of this study was to investigate association between 9p21.3 locus single nucleotide polymorphisms (SNP) and severity of coronary atheromatous burden. A total of 255 Caucasians (211 male, 44 female) with myocardial infarction younger 65 years old (mean 52,56±7,98 years) were enrolled. Genome DNA was extracted from venous blood using the phenol-chloroform extraction method. Two SNPs rs10757278 and rs1333049 (locus 9p21.3) were tested using real-time polymerase chain reaction (PCR) according to protocol (probes TaqMan, Applied Biosystems, 7900HT). The coronary angiograms were reviewed by independent angiographers who were blinded to the results of the genotype (Philips Allura Xper FD10). The total number of lesions, Gensini scores and SYNTAX score were derived. For the first time in males of Siberian population, genotype CC rs1333049 and genotype GG rs10757278 demonstrated a direct strong association with severity of coronary atheromatous burdenЦель исследования – изучить взаимосвязь локуса 9р21.3 с тяжестью коронарного атеросклероза в сибирской популяции. В исследование включены 255 больных (211 мужчин, 44 женщины) с инфарктом миокарда (ИМ) европеоидной расы в возрасте ≤ 65 лет (52,56±7,98 лет). Геномную ДНК выделяли из венозной крови фенол-хлороформным методом. Полиморфизм генов тестировали с помощью полимеразной цепной реакции (ПЦР) в режиме реального времени (зонды TaqMan, Applied Biosystems 7900HT). В исследовании изучались два однонуклеотидных полиморфизма (ОНП), расположенные в локусе 9p21.3 – rs10757278 и rs1333049. Оценка степени тяжести поражения коронарного русла производилась в заслепленном режиме в ходе стандартной полипроекционной коронарографии (Philips Allura Xper FD10), учитывались количество пораженных сегментов, интегральные индексы Gensini и SYNTAX. Впервые в сибирской популяции у мужчин доказана взаимосвязь генотипов GG rs10757278 и СС rs1333049 с тяжестью коронарного атеросклероз

Features of Peripheral Blood Th-Cell Subset Composition and Serum Cytokine Level in Patients with Activity-Driven Ankylosing Spondylitis

Th cells may exhibit pathological activity depending on the regulatory and functional signals sensed under a wide range of immunopathological conditions, including ankylosing spondylitis (AS). The relationship between Th cells and cytokines is important for diagnoses and for determining treatment. Accordingly, the aim of this study was to investigate the relationship between Th-cell subset composition and serum cytokine profile for patients with activity-driven AS. In our study, patients were divided into two groups according to disease activity: low-activity AS (ASDAS-CRP < 2.1) and high-activity AS (ASDAS-CRP > 2.1). The peripheral blood Th cell subset composition was studied by flow cytometry. Using multiplex analysis, serum cytokine levels were quantified and investigated. It was found that only patients with high-activity AS had reduced central memory (CM) Th1 cells (p = 0.035) but elevated numbers of CM (p = 0.014) and effector memory (EM) Th2 cells (p < 0.001). However, no activity-driven change in the Th17 cell subset composition was observed in AS patients. Moreover, low-AS activity patients had increased numbers of Tfh17 EM cells (p < 0.001), whereas high-AS activity was associated with elevated Tfh2 EM level (p = 0.031). The serum cytokine profiles in AS patients demonstrated that cues stimulating cellular immunity were increased, but patients with high-AS activity reveled increased IL-5 level (p = 0.017). Analyzing the data obtained from AS patients allowed us to conclude that Th cell subset differentiation was mainly affected during the CM stage and characterized the IL-23/IL-17 regulatory axis, whereas increased humoral immunity was observed in the high-AS activity group

Development of DNA aptamers for visualization of glial brain tumors and detection of circulating tumor cells

Here, we present DNA aptamers capable of specific binding to glial tumor cells in vitro, ex vivo, and in vivo for visualization diagnostics of central nervous system tumors. We selected the aptamers binding specifically to the postoperative human glial primary tumors and not to the healthy brain cells and meningioma, using a modified process of systematic evolution of ligands by exponential enrichment to cells; sequenced and analyzed ssDNA pools using bioinformatic tools and identified the best aptamers by their binding abilities; determined three-dimensional structures of lead aptamers (Gli-55 and Gli-233) with small-angle X-ray scattering and molecular modeling; isolated and identified molecular target proteins of the aptamers by mass spectrometry; the potential binding sites of Gli-233 to the target protein and the role of post-translational modifications were verified by molecular dynamics simulations. The anti-glioma aptamers Gli-233 and Gli-55 were used to detect circulating tumor cells in liquid biopsies. These aptamers were used for in situ, ex vivo tissue staining, histopathological analyses, and fluorescence-guided tumor and PET/CT tumor visualization in mice with xenotransplanted human astrocytoma. The aptamers did not show in vivo toxicity in the preclinical animal study. This study demonstrates the potential applications of aptamers for precise diagnostics and fluorescence-guided surgery of brain tumors.peerReviewe