Bulletin of RSMU

ISSN Print 2500–1094 ISSN Online 2542–1204

Bulletin of RSMU

BIOMEDICAL JOURNAL OF PIROGOV UNIVERSITY (MOSCOW, RUSSIA)

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Chicherin IV, Levitskii SA, Krasheninnikov IA, Tarassov I, Kamenski P

Mitochondrial DNA mutations cause severe inherited disorders in humans. To date, there are a few therapeutic strategies for their correction; however, it is highly unlikely that they would be routinely used in clinical practice. The past few years have witnessed the rapid progress of a genome editing technology known as CRISPR/Cas9. The present review focuses on the current strategies to combat mitochondrial mutations and reveals their major drawbacks. The article also explores the possibility of creating a possible specific CRISPR/Cas9 tool for correcting mitochondrial DNA mutations and provides a rough description of its mechanism of action. A particular focus is paid to technical challenges. On the whole, we see no principal barriers to implementing a mitoCRISPR/Cas9 system for treating mitochondrial disorders.

Received: 2017-06-20

Published online: 2017-07-19

DOI: 10.24075/brsmu.2017-03-05

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VIEWS 5160

Mogulevtseva YuA, Mezentsev AV, Bruskin SA

Matrix metalloproteinases play an important role in maintaining skin homeostasis, promote wound healing, and are involved in triggering inflammation. They are implicated in the structural changes occurring in the epidermis of psoriatic patients and also facilitate infiltration of the skin by immune cells by regulating permeability of dermal capillaries. In this light, control over the enzymatic activity of matrix metalloproteinases is crucial for a successful treatment outcome in patients with psoriasis. The aim of this work was to investigate the effect of RNA interference on the progression of psoriasis by targeting interstitial collagenase of epidermal keratinocytes. As part of the experiment, the latter were transduced with lentiviral particles that encode small hairpin RNA. Gene expression was measured by real time polymerase chain reaction. Enzymatic activity was measured by zymography. RNA interference was found to lead to a 20- and 4-fold decrease in the expression and enzymatic activity of interstitial collagenase, respectively. Expression of homologous genes (MMP2, -9 and -12) changed insignificantly. In contrast, there were marked changes in expression of cytokeratin (KRT1: 16.89 ± 0.97; KRT14: 2.36 ± 0.19; KRT17: 0.12 ± 0.01; KRT18: 0.56 ± 0.02), involucrin (0.79 ± 0.11) and filaggrin (6.99 ± 0.97). Besides, RNA interference caused a significant decline in cell migration rates, although it did not affect cell proliferation. Thus, small hairpin RNAs targeting interstitial collagenase are potentially therapeutic for psoriatic patients due to their ability to regulate expression of genes implicated in psoriasis (IVL, FLG, KRT1, -14 -17, and -18).

Received: 2017-02-17

Published online: 2017-07-19

DOI: 10.24075/brsmu.2017-03-04

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VIEWS 4337

Kuzevanova AYu, Luneva AS, Maslov MA, Karpukhin AV, Alimov AA

There are a number of problems that need to be addressed when designing an effective RNA interference-based drug including distribution of intravenously injected exogenous small interfering RNAs (siRNAs) in the organs and tissues of the patient. Insufficient data on siRNA distribution obtained using isotopic/fluorescent labeling offers no insight into whether the polymer retains its original structure after the injection. Quantitative real-time polymerase chain reaction that we used in our work provides a better response to the challenge. In our experiment LIVIN–specific siRNAs injected intravenously were distributed unevenly between tissues and their accumulation was dose-dependent (the study was conducted in mice using 2.5 and 7.5 mg/kg doses). Maximal accumulation was observed in the liver and spleen where siRNA concentration continued to increase between 48 and 96 hours after its administration. This demonstrates that the studied cationic lisosome/miRNA complex has long circulation time. We believe that the obtained data will be instrumental in finding an effective therapeutic dose, designing adequate regimens and preparing for preclinical or clinical trials of siRNA-based drugs.

Received: 2017-06-20

Published online: 2017-07-19

DOI: 10.24075/brsmu.2017-03-03

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VIEWS 4281

Vyakhireva JV, Filatova AYu, Krivosheeva IA, Skoblov MYu

RNA-interference enjoys a wide range of applications in medical and biological research. In particular, it is used to study functions of genes. One of the most popular approaches to this task is gene knockdown by small interfering RNA (siRNA). Currently there is no unified protocol for this method, which results in low reproducibility of experimental data. In the following article we outline the theoretical bases for this method and provide practical recommendations for its use in siRNA-mediated gene silencing experiments.

Received: 2017-06-25

Published online: 2017-07-19

DOI: 10.24075/brsmu.2017-03-02

Comments 0

VIEWS 5035

Filatova AYu, Sparber PA, Krivosheeva IA, Skoblov MYu

As of today, there have been about 2,500 gene therapy clinical trials initiated or completed worldwide. Some of the tested drugs have been already approved for clinical use. Most of these drugs target well-characterized protein-coding genes. At the same time, the past few years have witnessed an increasing interest in long noncoding RNAs (lncRNAs) and their role in cellular processes. Of 16,000 identified human lncRNA genes, biological functions have been elucidated for only two hundred. Nevertheless, we already know about their association with the development of 200 different disorders. In some cases these genes are the key element in disease pathogenesis, which makes long noncoding RNAs a promising target for gene therapy. To date, researchers successfully employ molecular biology techniques for the development of lncRNA-based therapeutic strategies. The following review focuses on the main approaches to gene therapy based on the use of lncRNA.

Received: 2017-06-25

Published online: 2017-07-18

DOI: 10.24075/brsmu.2017-03-01

Comments 0

VIEWS 4663

Guglya EB, Kotlobay AA, Sekretova EK, Volkova PV, Yampolsky IV

An extensive collection of plants gathered in the European part of Russia was screened for a substrate of fungal luciferase. This work was inspired by the recently discovered mechanism of bioluminescence in higher fungi and the structural similarity of fungal luciferin with some plant metabolites. Of all studied leaf extracts obtained from 200 different plants, bioluminescent activity was discovered in 10 species. Each of these species contained a plurality of active compounds. All the luminescent substrates were not identical to fungal luciferin (3-hydroxyhispidin) and were chemically unstable, rendering the attempt to isolate individual compounds for further structural characterization yet unsuccessful. This study is the first step towards engineering a self-luminescent plant based on a fungal enzyme-substrate bioluminescent system.

Received: 2017-02-19

Published online: 2017-06-04

DOI: 10.24075/brsmu.2017-02-10

Comments 0

VIEWS 4494

Malov IA, Strenev NV, Takhchidi KhP

Premacular hemorrhage occurs in various disorders and causes sudden unilateral or bilateral visual impairment. One of the well-established techniques to treat this condition is Ng:YAG laser hyaloidotomy. Below we report a case of premacular hemorrhage in the right and left eyes of a 23-year old patient with acute myeloblastic leukemia. Ng:YAG laser hyaloidotomy was successfully performed on both patient’s eyes at different puncture sites.

Received: 2017-04-10

Published online: 2017-06-02

DOI: 10.24075/brsmu.2017-02-09

Comments 0

VIEWS 4127

Tkacheva SV, Manapova ER, Sozinova YuM, Yakupova FM, Fazylova YuV

Interferon-based regimens for chronic hepatitis C (HCV) are quite common, just like interferon-free treatments, and are extensively used in Russia because interferon is widely available to most patients. In 2013 the original Russian drug cepeginterferon alpha-2b (cepegIFN alpha-2b marketed as Algeron by Biocad, Russia) was introduced into clinical practice. The aim of this study was to assess effectiveness and safety of cepegIFN alpha-2b as part of the combination therapy with ribavirin in patients with chronic HCV infection. The study was conducted over the period from 2014 to 2016 and recruited 37 patients with chronic genotype 1 HCV infection: 22 men and 15 women (mean age of 42.0 ± 5.2 years). All of them received the following combination antiviral therapy (AT): 1.5 μg/kg cepegIFN alpha-2b once a week and 15 μg/kg ribavirin daily over the period of 48 weeks. Effectiveness of AT was assessed by the rate of sustained virological response (SVR), i. e. aviremia achieved 24 weeks after the onset of treatment. In our SVR was observed in 26 patients (70.3 %). Adverse effects seen in the course of AT were typical of interferon-based drugs and ribavirin. CepegIFN alpha-2b dosage was corrected in two patients who developed neutropenia; ribavirin dosage was corrected in 3 patients who developed anemia. Based on the obtained results, we recommend including cepegIFN alpha-2b into the combination antiviral therapy in patients with chronic HCV infection.

Received: 2017-02-09

Published online: 2017-06-02

DOI: 10.24075/brsmu.2017-02-08

Comments 0

VIEWS 4239