Voprosy meditsinskoi khimii最新文献

The work represents an introduction article of editors of special issue of the magazine devoted to gene therapy and therapeutics. The main results of clinical gene therapy in the past decade are critically considered in connection with a changes of paradigms of the field. They are: 1) change of the main target of genetic therapy--correction of defects in chromosomes--onto expression and/or output of target genes for gene therapy; 2) transfer from gene transplantation to cell transplantation; 3) tendency for the use of safe/non-viral vectors instead of viral ones.; and 4) conflict of interests in gene therapy. Outlooks in the field are discussed.

Various aspects of use of specific interactions with a participation of carbohydrate and oligosaccharide ligands to increase an efficiency of gene transfer into eukaryotic cells (including in vivo experiments) are considered in details. Data on addressed gene delivery with applying carbohydrate-containing ligands (such as asialoglycoproteins and galactosides) are discussed in the paper. Results on the usage of glycoside ligands, containing lactose, mannose, glucose residues, for receptor-mediated gene transfer, are analysed. Special attention is paid to application of chitosans for functional gene transfer into eukaryotic cells, which is considered by authors as a case of receptor-mediated gene transfer. It is notice that neo-oligosaccharide vectors, recognizing surface lectins, represent very perspective type of gene delivery systems.

Gene therapy of chronic infectious diseases of urogenital tract represents a new perspective field in the modern biological and medical sciences. In the review discuss one of the new directions in gene therapy of urogenital infections caused by Mycoplasma: inhibition of mycoplasmal infection after administration of recombinant plasmid vectors, expressed the genes of cytotoxic peptides.

The paper highlights the new trends in gene therapy research area and clinical trials. It should be noted that the majority of firms involved in development of the scientific approaches to gene therapy are concentrated in the United States. The investments of the given companies on development and research of new genetic constructs also delivery systems make hundred millions dollars. The greatest part (more than 80%) of gene therapy clinical trials projects are also connected with the US research departments; the majority of them is related to tumor therapy. The advantages and drawbacks of the main methods of nucleic acids delivery to the cells are considered; diseases that are attempted to be treated using gene therapy methods are listed. A special attention of the review is devoted to the modern stand in research on cell and Duchenne muscular dystrophy (DMD) gene therapy, also brief description of basic results achieved in the authors laboratory is given. Basic original results of transfection of mdx mice (DMD biological models) with dystrophin cDNA delivered by gene gun, cationic liposomes, synthetic microspheres, viral olygopeptides and lactoferrine are summarized.

This review describes the systems of retroviral transfer and expression of the genes that are widely applied in basic biological research and in gene therapy. Unique features of retroviruses providing a background for construction of retroviral vectors and the methods to use these vectors are discussed.

The therapeutic potential of gene therapy in cardiovascular disease such as post-angioplasty restenosis, myocardial ischaemia and severe peripheral artery disease ischemia are considered.

Recent findings connected with in vivo use of artificial macromolecular complexes (genosomes) for functional gene transfer and delivery are discussed in the paper. Non-viral methods are the most safe for the purpose of human gene delivery. The cationic liposomes containing cholesterol are the most suitable for this purpose, because they possess high biodegradability and stability in blood stream. The DNA-liposome complexes should: (i) contain DNA in the condition at most protected from environmental influence, (ii) be rather homogeneous and of small size (40-80 nm). Injections of complexes into blood are the most effective; in a respect of organospecifity may be achieved by appropriate ligand selection. It is the most perspectively to increase the expression level by combining liposomes with viral peptides.

In the review the main advantages in development of the approaches to gene therapy of hereditary diseases are presented. Now more than 1000 genes of hereditary diseases are mapped and some hundreds are cloned which is prerequisite for gene therapy. The transfer of the recombinant gene into the cell and the subsequent expression of the transgene product are the rate-limiting steps for successful gene therapy. A variety of methods, including the use of physical methods, modified viruses and synthetic vectors, are currently being used in experiments and clinical trials. Since the approval and initiation of the first human gene therapy trial to treat ADA deficiency, there have been several dozen approved gene therapy trials but clear clinical result was stated for ADA deficiency only. Cystic Fibrosis, CF was among several hereditary diseases which were considered as a target for gene therapy. Experiments on development of recombinant gene constructions, gene delivery by adenovirus vectors and liposomes as well as by other constructions into epithelial lung cells, gene expression and on the safety of gene therapy procedures were relatively successful. Phase 1 gene therapy clinical trials of CF showed that some unaccounted physiological peculiarities of lung tissue of the patients diminished effectiveness of gene transfer, longevity of CFTR gene expression and in some cases unexpected immunological complications arises during clinical trials. Now an intensive attempt to overcome these problems in gene therapy of CF are undertaken.

The influence of thyroxine on the activities of enzymes of energy metabolism (hexokinase, 6-phosphofructokinase, pyruvate kinase, lactate dehydrogenase, glucose-6-phosphate dehydrogenase, NADP-isocitrate dehydrogenase, cytochrome c oxidase) was investigated in bone marrow myeloid cells and blood neutrophils of 3-10-day old neonatal piglets. Data obtained suggest different responsiveness of energy metabolism enzymes to thyroxine action. Repeated hormone injections resulted in the preferential stimulation of enzymes involved in oxidative stages of carbohydrate catabolism in animal myelocaryocytes, while the activities of anaerobic enzymes in these cells were less affected. At the same time glycolytic enzymes in neutrophil granulocytes showed higher sensitivity to thyroxine action than enzymes catalyzing oxidative stages of energy metabolism.

The induction of cytochrome P4501A1 (CYP1A1) enzyme activity reflects the increased ligand-dependent transcriptional activity of the cognate CYP1A1 gene. The list of ligands includes various xenobotics such as polycyclic and halogenated aromatic hydrocarbons. Until recently, similar role for endogenous compounds was unknown. In the present study the ability of the endogenous heme metabolite, bilirubin, to regulate CYP1A1 activity was examined. The following parameters were investigated: expression of CYP1A1 in rat liver at a level mRNA, protein and functional activity under at the experimental rising of blood bilirubin level. The influence of local ultrasound contact treatment of rats hepatic area in vivo (the intensity 0.4 W/cm2 and duration time of 10 minutes) on blood unconjugated bilirubin concentration and parameters of CYP1A1 transcriptional activity was also investigated. The ultrasound contact action on rat hepatic are increased blood unconjugated bilirubin concentration. The rise of bilirubin levels of in rat blood after intravenous administration of bilirubin as well as after ultrasound treatment was accompanied by increased mRNA CYP1A1, protein and functional activity of CYP1A1. The comparison of these data with that of time-dependent changes of parameters of CYP1A1 transcriptional activity under ultrasound action and experimental rising of blood bilirubin level suggest that induces CYP1A1 and may be an intermediate in the activation of CYP1A1 expression under ultrasound action.