- MeSH
- DNA, Antisense physiology ultrastructure MeSH
- RNA, Antisense physiology ultrastructure MeSH
- Gene Expression MeSH
- Publication type
- Review MeSH
Biologická terapie, jejímž mechanizmem účinku je použití monoklonálních protilátek proti nějakému proteinu, je používána v klinické praxi již řadu let. V současné době ale vstupují do klinické praxe nové léky ze skupiny biologické terapie, které účinkují na principu RNA-interference. RNA-interference je proces, kterým buňky všech živých organizmů regulují expresi svých genů a při kterém může být zastaven přenos informace o syntéze konkrétního proteinu mezi DNA a ribosomy. Pro terapeutické účely se tohoto efektu dosahuje podáním umělých syntetizovaných oligonukleotidů s přesně danou sekvencí nukleosidů. Jde buď o krátké úseky dvouvláknové RNA, nebo o jednovláknové oligonukleotidy. Pro klinické využití byla nutná pro zvýšení jejich stability a odstranění některých nežádoucích účinků jejich chemická modifikace, a dále pak vazba na další substance, které umožní jejich cílený transport do požadované tkáně. Celá řada těchto léků je již v pokročilých fázích klinických studií a některé z nich vstupují na farmaceutický trh.
Biological therapy, whose mechanism of action is the use of monoclonal antibodies against a protein, has been used in clinical practice for many years. However, new drugs from the group of biological therapies that act on the principle of RNA interference are now entering clinical practice. RNA interference is the process by which cells in all living organisms regulate the expression of their genes, and in which the transfer of information about the synthesis of a particular protein between DNA and ribosomes can be stopped. For therapeutic purposes, this effect is achieved by administering artificially synthesized oligonucleotides – short chains of RNA with a precise nucleoside sequence. These are either short stretches of double- stranded RNA or single-stranded oligonucleotides. For clinical use, their chemical modification was necessary to increase their stability and remove some of their side effects, and then binding to other substances to allow their targeted transport to the desired tissue. A number of these drugs are already in advanced stages of clinical trials, and some are entering the pharmaceutical market.
Effective translation of rare disease diagnosis knowledge into therapeutic applications is achievable within a reasonable timeframe; where mutations are amenable to current antisense oligonucleotide technology. In our study, we identified five distinct types of abnormal splice-causing mutations in patients with rare genetic disorders and developed a tailored antisense oligonucleotide for each mutation type using phosphorodiamidate morpholino oligomers with or without octa-guanidine dendrimers and 2'-O-methoxyethyl phosphorothioate. We observed variations in treatment effects and efficiencies, influenced by both the chosen chemistry and the specific nature of the aberrant splicing patterns targeted for correction. Our study demonstrated the successful correction of all five different types of aberrant splicing. Our findings reveal that effective correction of aberrant splicing can depend on altering the chemical composition of oligonucleotides and suggest a fast, efficient, and feasible approach for developing personalized therapeutic interventions for genetic disorders within short time frames.
- MeSH
- Oligonucleotides, Antisense * therapeutic use genetics MeSH
- Genetic Diseases, Inborn genetics therapy MeSH
- Humans MeSH
- Morpholinos therapeutic use genetics MeSH
- Mutation * MeSH
- RNA Splicing * MeSH
- Rare Diseases * genetics drug therapy MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
Previous reports from this laboratory have shown marked cytocidal effects of the ISIS-3466 antisense phosphorothioate oligodeoxynucleotide to the human nucleolar protein p120 on human cancer cell lines in vitro and inhibition of tumor growth in vivo in an i.p/i.p. LOX cell model (L. Perlaky et al. Anti-Cancer Drug Design 8:3-14, 1993). In this study, light and fluorescence microscopy showed that the number of LOX cells in mitosis decreased by 50% after incubation for 4 h in 0.2-0.4 microM antisense oligonucleotide; a 70% reduction in cell number was found from 8-72 h post-treatment. In addition, marked unravelling of nucleolar structures and chromatin fragmentation was found after a 4-h incubation. The nucleolar unravelling occurred in varying degrees ranging from partial unfolding to almost complete separation of the strands of nucleolar residues. Twenty four hours post-treatment, immunofluorescence staining with the anti-p120 monoclonal antibody showed reduced nucleolar protein p120 and translocation of the p120 protein from the nucleoli to the nucleoplasm. Analysis of the mechanisms of the nucleolar unravelling and inhibition of mitosis will provide further understanding of the cytocidal effects of the ISIS-3466 antisense oligonucleotide.
- MeSH
- Melanoma, Amelanotic * genetics chemistry pathology MeSH
- Antigens, Neoplasm * drug effects MeSH
- Oligodeoxyribonucleotides, Antisense * MeSH
- Oligonucleotides, Antisense pharmacology genetics MeSH
- Cell Nucleolus * drug effects MeSH
- Cell Nucleus drug effects MeSH
- Chromatin drug effects MeSH
- Microscopy, Fluorescence MeSH
- Nuclear Proteins * drug effects MeSH
- Humans MeSH
- Mitosis drug effects MeSH
- Molecular Sequence Data MeSH
- Tumor Cells, Cultured drug effects MeSH
- Base Sequence MeSH
- Thionucleotides pharmacology genetics MeSH
- Check Tag
- Humans MeSH
- Publication type
- Research Support, Non-U.S. Gov't MeSH
- MeSH
- Oligodeoxyribonucleotides, Antisense physiology genetics MeSH
- Financing, Government MeSH
- Genetic Therapy MeSH
- Humans MeSH
- RNA, Small Interfering physiology genetics MeSH
- MicroRNAs physiology genetics MeSH
- Gene Expression Regulation genetics immunology MeSH
- RNA Interference immunology MeSH
- Check Tag
- Humans MeSH
- MeSH
- RNA, Antisense genetics MeSH
- DNA-Binding Proteins genetics MeSH
- Phosphoproteins genetics MeSH
- Transcription, Genetic immunology MeSH
- Humans MeSH
- RNA, Neoplasm genetics MeSH
- Trans-Activators genetics MeSH
- Transcription Factors genetics MeSH
- Check Tag
- Humans MeSH
- Publication type
- Comparative Study MeSH
Sturgeons (chondrostean, acipenseridae) are ancient fish species, widely known for their caviar. Nowadays, most of them are critically endangered. The sterlet (Acipenser ruthenus) is a common Eurasian sturgeon species with a small body size and the fastest reproductive cycle among sturgeons. Such species can be used as a host for surrogate production; application is of value for recovery of critically endangered and huge sturgeon species with an extremely long reproductive cycle. One prerequisite for production of the donor's gametes only is to have a sterile host. Commonly used sterilization techniques in fishes such as triploidization or hybridization do not guarantee sterility in sturgeon. Alternatively, sterilization can be achieved by using a temporary germ cell exclusion-specific gene by a knockdown agent, the antisense morpholino oligonucleotide (MO). The targeted gene for the MO is the dead end gene (dnd) which is a vertebrate-specific gene encoding a RNA-binding protein which is crucial for migration and survival of primordial germ cells (PGCs). For this purpose, a dnd homologue of Russian sturgeon (Agdnd), resulting in the same sequence in the start codon region with isolated fragments of sterlet dnd (Ardnd), was used. Reverse transcription polymerase chain reaction confirmed tissue-specific expression of Ardnd only in the gonads of both sexes. Dnd-MO for depletion of PGCs together with fluorescein isothiocyanate (FITC)-biotin-dextran for PGCs labeling was injected into the vegetal region of one- to four-cell-stage sterlet embryos. In the control groups, only FITC was injected to validate the injection method and labeling of PGCs. After optimization of MO concentration together with volume injection, 250-μM MO was applied for sterilization of sturgeon embryos. Primordial germ cells were detected under a fluorescent stereomicroscope in the genital ridge of the FITC-labeled control group only, whereas no PGCs were present in the body cavities of morphants at 21 days after fertilization. Moreover, the body cavities of MO-treated and nontreated fish were examined by histology and in situ hybridization, showing gonads which had no germ cells in morphants at various stages (60, 150, and 210 days after fertilization). Taken together, these results report the first known and functional method of sturgeon sterilization.
- MeSH
- Oligonucleotides, Antisense * administration & dosage MeSH
- Cell Death MeSH
- Gene Knockdown Techniques methods veterinary MeSH
- Gonads chemistry MeSH
- DNA, Complementary chemistry MeSH
- Morpholinos * administration & dosage MeSH
- RNA-Binding Proteins analysis genetics MeSH
- Fishes * genetics MeSH
- Base Sequence MeSH
- Sequence Alignment MeSH
- Sterilization, Reproductive methods veterinary MeSH
- Germ Cells physiology MeSH
- Animals MeSH
- Check Tag
- Male MeSH
- Female MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
Cervical cancer is a killer disease that claims innumerable lives of women all over the world. Human Papilloma Virus (HPV) serotypes 16 and 18 have been implicated as the causal organism of the disease. Initiation of sexual intercourse at an early age, multiple sex partners, poor hygiene etc are regarded as risk factors that are also associated with progression of HPV infection. Two genes in HPV encode the oncoproteins E6 and E7. These proteins have been found to be in high content in cervical cancer positive cell lines. Attempts have been made to selectively inhibit the expression of these two oncogenes by molecular biological techniques. Of these RNA interference (RNAi) deserves special mention. RNAi occurs naturally in mammalian cells through which expression of a particular gene can be knocked down with high precision. Different authors from time to time have targeted to knock out these E6 and E7 proteins as a preventive measure of cervical cancer. The present review portrays the state of the art evaluation of retrospective as well as recent literature on these challenging techniques.
- MeSH
- DNA, Antisense MeSH
- Molecular Targeted Therapy MeSH
- Genetic Therapy MeSH
- Humans MeSH
- RNA, Small Interfering MeSH
- Uterine Cervical Neoplasms * genetics prevention & control therapy MeSH
- Oncogene Proteins, Viral * genetics MeSH
- Papillomaviridae genetics MeSH
- RNA Interference MeSH
- RNA, Catalytic MeSH
- Genes, Viral MeSH
- Check Tag
- Humans MeSH
- Female MeSH
- Publication type
- Review MeSH
