Vol 15, No 2 (2020)

The review presents the modern concept of the pathogenesis of diffuse alveolar damage, including acute respiratory distress sYndrome in coronavirus infection. It has been established that the so-called "cytokine storm”, which consists in the increased release of substances that are biologically active against the vascular wall and effector cells, leading to the progressive damage to endotheliocytes and alveolocytes, the development of alveolar and interstitial pulmonary edema with fatal respiratory failure and coagulopathy. An important factor in interstitial aggression is the appearance of autoreactive clones of plasma cells, dissemination of virusinfected leucocytes throughout the body with the involvement of various organs and systems, which exacerbates multiple organ failure. A poor prognosis for patients, the likelihood of developing pulmonary fibrosis after infection, according to several researchers, can be corrected by cell therapy. Allogeneic multipotent mesenchymal stromal cells (mesenchymal stem cells) are considered as first-line therapeutic cells. The accumulated experience of preclinical experiments made it possible to urgently proceed to conduct clinical trials of the safety of their use in patients with ARDS and to search for optimal indications to obtain maximum benefits for patients after transplantation. The combined efforts of many research groups can lead to reliable information on the cell therapy benefit and the need to include it in the standards of treatment of patients with this extremely severe pathology.

Epicardium is actively involved in the embryonic heart development and its repair after injury, which allows it to be considered as a potential target for the treatment of heart diseases. In this regard, the study of the mechanisms of its development, the components of the microenvironment, as well as the signals regulating the behavior of epicardial progenitor cells, is the most important area of modern cardiology. This review considers the results of recent studies of homeostasis of epicardial cells and technological advances to modulate their activity, which is essential for the development of new therapeutic agents.

New effective and easy-to-use genome editing tools open up the possibility of treating various diseases, including hereditary, oncological and infectious. The review provides information about clinical trials of developing therapies for monogenic diseases using genome editing systems such as zinc finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs), and clustered regularly interspaced short palindromic repeats and CRISPR-associated protein 9 (CRISPR/Cas9). Clinical trials are carried out for six hereditary diseases: mucopolysaccharidosis types I and II, sickle cell anemia, β-thalassemia, hemophilia B, and Leber congenital amaurosis 10. Knowledge about safety and efficacy of genome editing are still limited and the long-term effects of such intervention have to be studied.

Optimization of the reparative regeneration of striated skeletal muscle tissue is actual for clinical practice. Volumetric muscle loss usually heals through the fibrous scar formation. Herein, there are numerous of methods under developed focused on reparative myogenesis induction. One of the promising approaches in this area is formed by gene-activated materials, particularly, in the hydrogel form. We developed a gene-activated hydrogel based on hyaluronic acid and plasmid DNA with the gene of vascular endothelial growth factor A (VEGF-A). Firstly, we showed a biocompatibility of the product in the subcutaneous test in mice. Using marker plasmid DNA carrying the luciferase gene, prolonged delivery of gene constructs to cells in vivo with a peak in transgene expression at day 7 was confirmed, while the same plasmid DNA in an aqueous solution provided a maximum level of delivery at day 1. Being implanted into a volumetric defect of the anterior tibial muscle in rats the gene-activated hydrogel activated angiogenesis in 2 weeks after surgery and induced MYH7B+-muscle fibers formation in the central zone of the defect at average number 50,0±16,1 and 21,8±10,5 in 2 and 4 weeks, respectively, whereas a hydrogel without plasmid DNA did not have any myogenic effects. Thus, plasmid DNA with VEGFA in the sodium alginate-based hydrogel induced angiogenesis in the volumetric muscle loss model and stimulated reparative myogenesis that could be used for further development of products effective for treatment of patients with muscle pathology.

Chronic inflammation and oxidative stress play key role in arterial wall changes and cardiovascular diseases. There is limited evidence on influence of genotypes, which are correlated with dironic inflammation and oxidative stress at arterial wall changes (pulse wave velocity, carotid artery intima-media thickness, endothelium-dependent vasodilation, presence of atherosclerotic plaques) and risk factors of cardiovascular diseases. We examined association of TNF-238G>A polymorphism, MMP9 -1562C>T polymorphism, CYBA c.214Т>С polymorphism with arterial wall changes and risk factors of cardiovascular diseases in 160 healthy people of different ages. GG genotype of TNF -238G>A polymorphism was associated with lower levels of aldosterone (p=0,021), higher levels of glycated haemoglobin (p=0,02) and insulin-like growth factor (p=0,032). СТ genotype of MMP9 -1562C>T polymorphism was associated with most commonly found obesity (p=0,05). CC and TC genotypes of CYBA c.214Т>С polymorphism were associated with shorter leucocyte telomere length (p=0,011). There wasn't found any association of TNF, MMP9, CYBA polymorphism with arterial wall changes. Association was found between TNF -238G>A polymorphism and MMP9 -1562C>T polymorphism with metabolic parameters, CYBA c.214Т>С polymorphism with leucocyte telomere length.

The effectiveness of the in vitro fertilization (IVF) program does not exceed 40% and mostly depends on the oocytes quality, that is affected by the composition of the follicular fluid: a content of cytokines, growth factors, cell-free DNA (cfDNA), et al. Increased level of cfDNA in the follicular fluid is associated with indicators of ovarian reserve, as well as the effectiveness of stimulation in the IVF program. One possible reason for the high level of cfDNA can be considered an increase of IL-8 concentration. However, the role of IL-8 in regulation of reproductive processes is ambiguous and is presented in a few studies. In this connection, the aim of the study was to investigate a content of cfDNA and IL-8 in the follicular fluid of women, as well as the relationship between them, depending on the parameters of folliculo- and oogenesis, early embryogenesis and IVF outcomes. 62 women with infertility and undergoing IVF treatment have been enrolled in the study. We collected follicular fluid samples from dominant follicles using transvaginal ultrasound aspiration. The concentration of IL-8 in the follicular fluid was evaluated by flow fluorimetry. Measurement of cfDNA in the follicular fluid was performed by fluorimetric method. A negative correlation between IL-8 level in the follicular fluid and the number of received oocytes, as well as the quality of embryos was revealed. At the same time, a higher level of cfDNA was recorded in women with low blastocyst quality and non-developing pregnancy. Correlation analysis showed the absence of a significant direct relationship between IL-8 and cfDNA in the whole group. Moreover, in subgroups characterized by a higher IL-8 level, we found a weak negative correlation between cfDNA and IL-8 concentrations. The results suggest that enlarged level of cfDNA in the follicular fluid is not a consequence of IL-8 increase; the levels of IL-8 and cfDNA in the follicular fluid are two independent factors with multidirectional effects involved in various stages of reproductive process.

The environment for the development of products containing live cells as an active substance is being set up in Russia for several years. However, the acceptable regulatory framework, which would have allowed to develop and place cell therapy products on the market, complying with international requirements for safety, efficacy, and quality, has not been yet established. Moreover, in addition to the national legal framework, the Eurasian Economic Union regulatory model has emerged that introduces in parallel a more straightforward pathway for the development and clinical use of this important medical product category. In this paper we address the domestic and Eurasian regulatory systems for products containing live cells as an active ingredient and suggest certain possible approaches to support this area of biotechnology.