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Vol 18, No 2 (2023)
Reviews
Contribution of signaling pathways associated with cellular senescence and regulatory non-coding RNAs to chronic obstructive pulmonary disease
Abstract
Chronic obstructive pulmonary disease (COPD) is a multifactorial disease of the respiratory system that affects the lung parenchyma and airways; it is one of the leading causes of death in the world, which explains the constant search for new approaches to diagnosis, treatment and prevention of the disease. COPD develops as a result of a complex interaction of molecular genetic factors, a network of epigenetic regulators, and environmental factors that are closely related to lifestyle. The molecular pathogenesis of COPD may include the mechanisms of alteration of the regulation of stressful reactions that prevent cellular senescence.
Non-coding RNAs play an important role in the regulation of various intracellular signaling pathways and is the most relevant subject of genetic studies of various pathological phenotypes. The expression profile of long non-coding RNA is often disregulated in various diseases. Information on the role of long non-coding RNA in the development of COPD is limited. Long non-coding RNA and the target-genes of signaling pathways involved in cellular senescence form a complex interactive network and may be targets for disease therapy.
The review presents the data concerning some aspects of the molecular pathogenesis of COPD, as well as role of long non-coding RNAs in the development of the COPD.
93-108
Pulmonary fibrosis: risk factors, pathogenesis and in vivo/in vitro experimental modeling
Abstract
Pulmonary fibrosis (PF) is a group of lung diseases characterized by scar formation and interstitial pneumonia with a mean life expectancy of 3–5 years’post diagnosis. Risk factors for developing PF include external (environment) and internal (genetic risk factors). The central role in the formation of fibrosis is played by the massive myofibroblasts and the excessive deposition of extracellular matrix, including collagen I type.
The main approaches for PF treatment is either lung transplantation or therapeutic treatment by antifibrotic drugs. However, for both approaches there are a number of limitations: for surgical — the lack of donor organs and immune rejection of transplanted tissues, for therapeutic — the drugs that are identified in animal studies fail in human clinical trials. Thus, there is a necessity for advancing of humanized in vitro models to improve treatments prior to human clinical trials.
The development of different tissue (two-, three-dimensional) models has created systems capable of emulating human lung structure, function, and cell and matrix interactions, which have shown potential for in vitro drug testing. In this review, we focused on PF risk factors, development mechanisms, and a review of the main in vitro and in vivo models for studying PF.
109-121
Original Study Articles
Analysis of the results of transplantation of the retinal pigment epithelium in the experiment
Abstract
BACKGROUND: A promising method for treating the pathology of the retinal pigment epithelium in age-related macular degeneration is cell replacement therapy.
AIM: The aim of the study was to analyze the results of cell transplantation in the form of a cell suspension into the subretinal space at various times.
MATERIALS AND METHODS: The material of the study was 20 rabbits (40 eyes) of the New Zealand albino breed. A month after the modeling of retinal pigment epithelium atrophy and retinal degeneration, rabbits underwent subretinal transplantation of induced retinal pigment epithelium in the form of a cell suspension. Optical coherence tomography and autofluorescence studies were conducted in a period of up to 8 months. Enucleated eyes of animals were subjected to morphological study.
RESULTS: When observing rabbits with a previously created model of atrophy in the long term, it was found that the cells of the transplanted retinal pigment epithelium remained viable for the entire period. There were no inflammatory reactions from the eyeball, clouding of the optical media, pathological changes in the structure of the retina.
CONCLUSION: Thus, with the introduction of a suspension of induced retinal pigment epithelium cells with atrophy of the retinal pigment epithelium, the injected cells retain their viability for up to 8 months.
123-132
Evaluation of in vitro functional activity of ANB4 drug for the treatment of spinal muscular atrophy
Abstract
BACKGROUND: Spinal muscular atrophy (SMA) is a monogenic neurodegenerative disease. SMA is caused by a deficiency of the functional survival motor neuron protein (SMN) as a result of a mutation in the SMN1 gene. BIOCAD is developing a domestic gene therapy drug for the treatment of SMA based on recombinant adeno-associated virus serotype 9 (rAAV9) carrying the SMN1 gene (ANB4). In vitro evaluation of the functional activity of ANB4 will allow a more complete characterization of the drug.
AIM: Development of an accurate and reproducible in vitro functional test that reflects the clinical mechanism of action of ANB4.
METHODS: To model SMA, the SMN1 gene was knocked down by transfection with small interfering RNA. Amount of the SMN protein was measured by enzyme-linked immunosorbent assay. The functional activity of the drug was evaluated by analysis of Gemin2 protein level using the western blot analysis.
RESULTS: Analytical method has been developed to assess the functional activity of the ANB4 drug for the treatment of SMA type 1 (rAAV9 carrying the SMN1 gene). The developed technique made it possible to obtain accurate and reproducible results. Production of Gemin2 after knockdown of SMN1 and the introduction of exogenous SMN1 gene was restored to control values, comparable with the restoration of the level of the SMN protein.
CONCLUSION: The developed technique closely reflects the clinical mechanism of action of the ANB4 drug for the treatment of SMA. By evaluating the functional activity in vitro, accurate and reproducible results were obtained in accordance with the required standards, including the principles of 3R.
133-143
Comparative analysis of the regenerative potential of blood derivatives on a cell model of corneal stromal injury
Abstract
BACKGROUND: Agents based on blood derivatives contain substances that accelerate the healing of wound defects. Nowadays data have been accumulated that various blood derivatives have different effects on regeneration processes. Therefore, a comparative study of the effect of several types of such preparations on the corneal stroma’s cell culture is relevant.
AIM: Impact assessment of 3 different blood derivatives on the processes of proliferation, migration and cell death of keratocytes in vitro.
MATERIAL AND METHODS: This study was conducted on a primary cell culture of human corneal keratocytes. Blood samples were obtained from the cubital vein of healthy volunteers after signing informed consent to participate in the study. Monolayer wound healing test, formazan test to assess proliferation, staining for Annexin V to assess the processes of apoptosis and necrosis were perfomed. Moreover, effect evaluation of platelet-rich plasma, intact serum and processed serum were performed.
RESULTS: It was shown that all 3 stimulants have a positive effect on the regeneration of the experimental stromal defect, but it is implemented in different ways. Processed whey showed the greatest stimulating effect on the process of cell migration. This serum reduced the level of cell death, but had almost no effect on proliferation processes. It is advisable to further study different types of stimulants to obtain processed serum. Intact serum increased the percentage of dividing cells more than others, but also proportionally increased the percentage of death in culture. Platelet-rich plasma showed the weakest effect on the migratory ability of keratocytes, but it still differed significantly from the control. This stimulator had little effect on the process of cell division, and had no effect on the overall level of cell death.
CONCLUSION: Experimental data can be taken into account in choosing of the management in clinical practice and personalized usage of blood derivatives, specifically in prescribing some blood derivatives to accelerate regeneration, and other derivatives for reducing cell death’s processes.
145-153
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