RNA therapeutics inhibit the expression of specific proteins/RNAs by targeting complementary sequences of corresponding genes or encode proteins for the synthesis desired genes to treat genetic diseases. RNA-based therapeutics are categorized as oligonucleotide drugs (antisense oligonucleotides, small interfering RNA, RNA aptamers), and mRNA drugs. The antisense oligonucleotides and small interfering RNA for treatment of genetic diseases have been approved by the FDA in the United States, while RNA aptamers and mRNA drugs are still in clinical trials. Chemical modifications can be applied to RNA drugs, such as pseudouridine modification of mRNA, to reduce immunogenicity and improve the efficacy. The secure and effective delivery systems such as lipid-based nanoparticles, extracellular vesicles, and virus-like particles are under development to address stability, specificity, and safety issues of RNA drugs. This article provides an overview of the specific molecular mechanisms of eleven RNA drugs currently used for treating genetic diseases, and discusses the research progress of chemical modifications and delivery systems of RNA drugs.
Aptamers, Nucleotide ; RNA, Small Interfering/therapeutic use* ; RNA, Messenger ; Oligonucleotides, Antisense/therapeutic use*

Antisense oligonucleotides (ASODN) for therapy is a genetic technology which is based on the base-complementary principle. DNA or RNA sequence synthesized by biotechnology is transferred into the target cells to form mRNA-DNA or mRNA-RNA double strand for inhibiting the expression of target genes. In this way we can control and treat some diseases. The development of antisense oligonucleotides drugs has opened a new area of genetic pharmacology. This paper reviews its classifications, mechanics and its wide application in the treatment of viral infection, tumor and cardiovascular diseases. At the same time we pose the problems that need solving.
Arterial Occlusive Diseases ; therapy ; Humans ; Neoplasms ; therapy ; Oligonucleotides, Antisense ; pharmacology ; therapeutic use ; Virus Diseases ; therapy

Cell Line, Tumor ; Genes, ras ; Humans ; Oligonucleotides, Antisense ; therapeutic use ; Pancreatic Neoplasms ; genetics ; therapy ; Point Mutation ; RNA, Messenger ; analysis

In this paper, the situation on antisense oligonucleotide as a means of gene therapy was outlined, and the main factors impeding its progress at present was summarized. The one of main factors is the efficiency of antisense oligonucleotide as a drug and the other is the side-effect in clinical use. At the level of cell and gene, these influential factors were analyzed in detail. The main factor that makes side-effect in using antisense oligonucleotide is the difficulty to distinguish effectively homologous-gene from target gene. The another factor is the secondary structure and three-dimensional structure of target gene that seriously affect antisense oligonucleotide to arrive at target position. The third problem is what can affect antisense oligonucleotide transmission and quick annealing. How use computer technique to analyze fully the target gene of antisense oligonucleotide including the secondary structure and homology of target gene, and to design effective antisense oligonucleotide, in order to reduce its side-effect in clinical use of antisense oligonucleotide as a drug of gene therapy, and the computer-aid design method were described.
Computer-Aided Design ; Fusion Proteins, bcr-abl ; genetics ; Genetic Therapy ; Humans ; Nucleic Acid Conformation ; Oligonucleotides, Antisense ; therapeutic use ; RNA ; chemistry

MicroRNAs (miRNAs) are a class of highly abundant non-coding RNA molecules that are involved in several biological processes. Many miRNAs are often deregulated in several diseases including cancer. There is substantial interest in exploiting miRNAs for therapeutic applications. In this editorial, we briefly review current advances in the use of miRNAs or antisense oligonucleotides (antagomirs) for such therapies. One of the key issues related to therapy using miRNAs is degradation of naked particles in vivo. To overcome this limitation, delivery systems for miRNA-based therapeutic agents have been developed, which hold tremendous potential for improving therapeutic outcome of cancer patients.
Drug Delivery Systems ; methods ; Gene Expression Regulation, Neoplastic ; Genetic Therapy ; Humans ; MicroRNAs ; genetics ; metabolism ; therapeutic use ; Neoplasms ; genetics ; metabolism ; therapy ; Oligonucleotides, Antisense ; therapeutic use

Bcr-abl antisense oligodeoxynucleotides (AS-ODNs) have provided evidence of an antileukemia effect when tested in vitro against Philadelphia-positive cells. In order to investigate the efficacy of AS-ODNs as purging agents in chronic myeloid leukemia (CML) patients, K562 cells, a human CML cell line, were treated in vitro with various types of AS-ODNs and interferon-alpha. Cells were treated in vitro for 0 and 36 hr with 40 microgram/mL of AS-ODNs, respectively, and incubated at 37 degrees C for 36 hr. Cytotoxic effects were measured by counting the number of viable cells as well as by MTT test. Clonogenic activities were evaluated by methylcellulose culture for 2 weeks. The effects of purging agents on the rearrangement of bcrabl gene were evaluated by RT-PCR. AS-ODNs inhibited the proliferation of K562 cells with time in cell count assay and MTT test. AS-ODNs were superior to INF-alpha in inhibiting clonogenic activity (recovery rate; 26.3% vs 64.0%). After incubation with bcr-abl AS-ODNs primers and mRNA isolated from K562 cells, positive bands were abolished, especially of b3a2 type and phosphorothioate type. Our results suggest that AS-ODNs mediated purging may be one of the efficient methods and that autograft may be an alternative treatment for allograft in high-risk group patients of CML if they do not have a stem cell donor.
Bone Marrow Purging* ; Colony-Forming Units Assay ; Fusion Proteins, bcr-abl/genetics ; Fusion Proteins, bcr-abl/metabolism ; Hematopoietic Stem Cells/physiology* ; Human ; Leukemia, Myeloid, Chronic/therapy ; Neuroblastoma/therapy* ; Oligonucleotides, Antisense/metabolism* ; Oligonucleotides, Antisense/therapeutic use ; Transplantation, Autologous* ; Tumor Cells, Cultured

Angiogenesis Inhibitors ; therapeutic use ; Animals ; Gene Transfer Techniques ; Genetic Therapy ; trends ; Genetic Vectors ; Humans ; Immunotherapy ; Oligonucleotides, Antisense ; therapeutic use ; Pancreatic Neoplasms ; drug therapy ; therapy ; Prodrugs ; therapeutic use ; RNA, Catalytic ; therapeutic use

OBJECTIVETo define changes in clusterin expression following short-term neoadjuvant hormone therapy (NHT) and its biological significance in prostate cancer tissues.

METHODSTwenty-six archival radical prostatectomy (RP) specimens without receiving NHT, 19 needle biopsies and corresponding 19 RP specimens following 3-month NHT, were subjected to immunohistochemical clusterin staining.

RESULTSStaining for clusterin was mainly found in cytoplasm and part of extracellular matrix. Clusterin expression was significantly greater in RP specimens with preoperative NHT (t = 2.91, P < 0.01); Needle biopsies obtained before NHT consistently demonstrated lower staining intensity (1.42 +/- 0.51) than corresponding RP specimens (2.16 +/- 0.60) following 3-month NHT (t = 7.10, P < 0.01).

CONCLUSIONSUpregulation of clusterin in part accounts for malignant progression of prostate cancer through its anti-apoptotic action following androgen withdrawal. These findings support that adjuvant therapy targeting clusterin may enhance androgen ablation therapy in advanced prostate cancer.

Aged ; Clusterin ; genetics ; metabolism ; Humans ; Immunohistochemistry ; Male ; Middle Aged ; Neoadjuvant Therapy ; methods ; Oligonucleotides, Antisense ; therapeutic use ; Prostatic Neoplasms ; metabolism ; pathology ; therapy

With the proceeding of the human genome program(HGP), new genes related to male reproduction have been cloned continuously by different methods. However, investigation of the gene function is still at the rudimentary stage. In spite of the advances in the common methods of studying gene function, some special methods have yet to be developed concerning the study of the genes related to male reproduction. This paper summarizes the current methods of study on gene function relevant to male reproduction.
Animals ; Gene Targeting ; Gene Transfer, Horizontal ; Genes ; physiology ; Genetic Vectors ; genetics ; Humans ; Male ; Models, Animal ; Oligonucleotides, Antisense ; therapeutic use ; Reproduction ; genetics ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVETo investigate the synergistic effects of antisense HIF-1alpha gene therapy combined with B7-1-mediated immunotherapy on cancer treatment.

METHODSAntisense HIF-1alpha and B7-1 expression vector were constructed. Lymphoma cells EL-4 were injected subcutaneously into C57BL/6 mice and transplanted lymphomas were established. The mice received either antisense HIF-1alpha, B7-1, or a combinational agent, complexed with DOTAP cationic liposomes. The tumor growth in the mice was monitored. Expression of HIF-1alpha, B7-1 and VEGF were detected by immunohistochemistry and Western blotting. The tumor blood vessels were immunostained with CD31- antibodies and the tumor vascular density was assessed by light microscopy.

RESULTSGene transfer of plasmid expressing the encoded antisense HIF-1alpha inhibited VEGF expression and reduced vascular density in the tumors, eradicated tumors in diameter smaller than 0.1 cm and only retarded the growth of larger tumors. Whereas combination of antisense HIF-1alpha gene therapy and B7-1 immunotherapy eradicated all tumors in diameter of 0.4 cm.

CONCLUSIONAntisense HIF-1alpha blocks tumor hypoxia pathway by downregulating VEGF expression, reduction of vascular density and enhances B7-1-mediated immunotherapy. Strategies that target HIF-1 may have therapeutic potential in cancer treatment and are worthy of further studying.

Animals ; B7-1 Antigen ; genetics ; therapeutic use ; Gene Transfer Techniques ; Genetic Therapy ; methods ; Hypoxia-Inducible Factor 1, alpha Subunit ; genetics ; therapeutic use ; Lymphoma ; immunology ; therapy ; Male ; Mice ; Mice, Inbred C57BL ; Neovascularization, Pathologic ; therapy ; Oligonucleotides, Antisense ; genetics ; therapeutic use