Antisense & nucleic acid drug development最新文献_第10页

Increased UV light sensitivity in transgenic Drosophila expressing the antisense XPD homolog. 表达反义XPD同源基因的转基因果蝇对紫外光的敏感性增加。

Antisense & nucleic acid drug development Pub Date : 2001-04-01 DOI: 10.1089/108729001750171399

M. T. Sandoval, M. Zurita

引用次数: 4

Tissue distribution and physiologically based pharmacokinetics of antisense phosphorothioate oligonucleotide ISIS 1082 in rat. 反义硫代寡核苷酸ISIS 1082在大鼠体内的组织分布及生理药代动力学。

Antisense & nucleic acid drug development Pub Date : 2001-02-01 DOI: 10.1089/108729001750072092

B. Peng, J. Andrews, I. Nestorov, B. Brennan, P. Nicklin, M. Rowland

{"title":"Tissue distribution and physiologically based pharmacokinetics of antisense phosphorothioate oligonucleotide ISIS 1082 in rat.","authors":"B. Peng, J. Andrews, I. Nestorov, B. Brennan, P. Nicklin, M. Rowland","doi":"10.1089/108729001750072092","DOIUrl":"https://doi.org/10.1089/108729001750072092","url":null,"abstract":"The aim of this study was to develop a whole body physiologically based model of the pharmacokinetics (PBPK) of the phosphorothioate oligonucleotide (PS-ODN) ISIS 1082 in vivo. Rats were administered an intravenous (i.v.) bolus dose of ISIS 1082 (10 mg/kg plus 3H tracer), and arterial blood and tissues were taken at specific times up to 72 hours. Radioactivity was measured in all samples. The parent compound was determined specifically in blood and tissues at 90 minutes and in liver and kidney also at 24 hours, using capillary gel electrophoresis (CGE). A whole body PBPK model was fitted to the combined blood and tissue radioactivity data using nonlinear regression analysis. CGE analysis indicated that the predominant species in plasma and all tissues is ISIS 1082, together with some n-1 and n-2 metabolites. Total radioactivity primarily reflects these species. The whole body model successfully described temporal events in all tissues. However, to adequately model the experimental data, all tissues had to be partitioned into vascular and extravascular spaces to accommodate the relatively slow distribution of ISIS 1082 out of blood because of a permeability rate limitation. ISIS 1082 distributes extensively into tissues, but the relative affinity varies enormously, being highest for kidney and liver and lowest for muscle and brain. A whole body PBPK model with a permeability rate limited tissue distribution was developed that adequately described events in both blood and tissue for an oligonucleotide. This model has the potential not only to characterize the events in individual tissues throughout the body for such compounds but also to scale across animal species, including human.","PeriodicalId":7996,"journal":{"name":"Antisense & nucleic acid drug development","volume":"15 1","pages":"15-27"},"PeriodicalIF":0.0,"publicationDate":"2001-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"89812403","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 39

Introduction of an RNA stability element at the 5'-end of an antisense RNA cassette increases the inhibition of target RNA translation. 在反义RNA盒的5'端引入RNA稳定元件增加了靶RNA翻译的抑制作用。

Antisense & nucleic acid drug development Pub Date : 2001-02-01 DOI: 10.1089/108729001750072100

H. Engdahl, Magnus Lindell, E. Wagner

{"title":"Introduction of an RNA stability element at the 5'-end of an antisense RNA cassette increases the inhibition of target RNA translation.","authors":"H. Engdahl, Magnus Lindell, E. Wagner","doi":"10.1089/108729001750072100","DOIUrl":"https://doi.org/10.1089/108729001750072100","url":null,"abstract":"This communication describes improvement strategies used on a previously described two-unit antisense RNA cassette system. This cassette system encodes RNA with noncontiguous regions of complementarity to a bacterial target RNA, lacI mRNA. One of the units of complementarity was contained within an RNA stem-loop resembling that of the very efficient, naturally occurring antisense RNA CopA. As relatively low inhibitory activity was obtained previously, we tested variants in which several stem-loops were combined within one RNA, each of them directed against a different stretch of target RNA. One to four stem-loop RNAs were tested and found to be relatively ineffective, likely because of low metabolic stability. To increase the intracellular stability of these and other antisense RNAs, a stabilizer element (stem-loop derived from gene 32 mRNA of phage T4) was inserted at their 5'-ends. The results indicate that addition of this element indeed increased antisense RNA efficiency in vivo. As expected, this effect was primarily due to a longer antisense RNA half-life, as shown by RNA abundance (Northern analysis) and decay rates (rifampicin runout experiments). In summary, the results reported indicate that rational design of antisense RNA is feasible, but that the degree of inhibition (approximately 75% maximum inhibition) accomplished here could still be improved.","PeriodicalId":7996,"journal":{"name":"Antisense & nucleic acid drug development","volume":"15 1","pages":"29-40"},"PeriodicalIF":0.0,"publicationDate":"2001-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"83098789","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 19

Complex host cell responses to antisense suppression of ACHE gene expression. 复杂宿主细胞对ACHE基因表达反义抑制的反应。

Antisense & nucleic acid drug development Pub Date : 2001-02-01 DOI: 10.1089/108729001750072128

N. Galyam, D. Grisaru, M. Grifman, N. Melamed‐Book, F. Eckstein, S. Seidman, A. Eldor, H. Soreq

{"title":"Complex host cell responses to antisense suppression of ACHE gene expression.","authors":"N. Galyam, D. Grisaru, M. Grifman, N. Melamed‐Book, F. Eckstein, S. Seidman, A. Eldor, H. Soreq","doi":"10.1089/108729001750072128","DOIUrl":"https://doi.org/10.1089/108729001750072128","url":null,"abstract":"3'-End-capped, 20-mer antisense oligodeoxynucleotides (AS-ODN) protected with 2'-O-methyl (Me) or phosphorothioate (PS) substitutions were targeted to acetylcholinesterase (AChE) mRNA and studied in PC12 cells. Me-modified AS-ODN suppressed AChE activity up to 50% at concentrations of 0.02-100 nM. PS-ODN was effective at 1-100 nM. Both AS-ODN displayed progressively decreased efficacy above 10 nM. In situ hybridization and confocal microscopy demonstrated dose-dependent decreases, then increases, in AChE mRNA. Moreover, labeling at nuclear foci suggested facilitated transcription or stabilization of AChE mRNA or both under AS-ODN. Intracellular concentrations of biotinylated oligonucleotide equaled those of target mRNA at extracellular concentrations of 0.02 nM yet increased only 6-fold at 1 microM ODN. Above 50 nM, sequence-independent swelling of cellular, but not nuclear, volume was observed. Our findings demonstrate suppressed AChE expression using extremely low concentrations of AS-ODN and attribute reduced efficacy at higher concentrations to complex host cell feedback responses.","PeriodicalId":7996,"journal":{"name":"Antisense & nucleic acid drug development","volume":"4 1","pages":"51-7"},"PeriodicalIF":0.0,"publicationDate":"2001-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"87192987","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 23

Impact of a Phosphorothioate Oligodeoxynucleotide MCP-1 on NF-κB, AP-1, SP1 and NF-κB, and AP-1 Subunit Composition in Human Pulmonary Endothelial Cells 硫代寡脱氧核苷酸MCP-1对人肺内皮细胞NF-κB、AP-1、SP1和NF-κB以及AP-1亚基组成的影响

Antisense & nucleic acid drug development Pub Date : 2001-02-01 DOI: 10.1089/108729001750072137

U. Maus, W. Seeger, J. Lohmeyer

引用次数: 3

Development of ribozymes that target stathmin, a major regulator of the mitotic spindle. 以有丝分裂纺锤体的主要调节因子安定素为目标的核酶的发展。

Antisense & nucleic acid drug development Pub Date : 2001-02-01 DOI: 10.1089/108729001750072119

S. Mistry, C. Benham, G. Atweh

{"title":"Development of ribozymes that target stathmin, a major regulator of the mitotic spindle.","authors":"S. Mistry, C. Benham, G. Atweh","doi":"10.1089/108729001750072119","DOIUrl":"https://doi.org/10.1089/108729001750072119","url":null,"abstract":"Stathmin is a major cytosolic phosphoprotein that plays an important role in the control of cellular proliferation by regulating the dynamics of the microtubules that make up the mitotic spindle. Because stathmin is expressed at high levels in all human cancers, it is an attractive molecular target for anticancer interventions. We had shown previously that antisense stathmin inhibition results in marked abrogation of the transformed phenotype of leukemic cells in vitro and in vivo. Unlike the antisense approach, ribozymes can catalytically cleave several molecules of target RNA. This may provide a more efficient strategy for downregulating genes, such as stathmin, that are expressed at very high levels in cancer cells. We designed several antistathmin hammerhead ribozymes and tested their cleavage activity against short synthetic stathmin RNA substrates. In vitro cleavage studies demonstrated site-specific cleavage of stathmin RNA that was dependent on ribozyme concentration and duration of exposure to ribozyme. The most active antistathmin ribozyme was capable of cleaving >90% stathmin RNA in a catalytic manner, cleaving multiple substrate molecules per ribozyme molecule. We also demonstrated that the designed antistathmin ribozymes are capable of selectively cleaving native stathmin RNA in a mixture of total RNA isolated from leukemic cells. These antistathmin ribozymes may provide a novel and effective form of gene therapy that may be applicable to a wide variety of human cancers.","PeriodicalId":7996,"journal":{"name":"Antisense & nucleic acid drug development","volume":"82 1","pages":"41-9"},"PeriodicalIF":0.0,"publicationDate":"2001-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"81236034","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 22

Improved intracellular delivery of oligonucleotides by square wave electroporation. 通过方波电穿孔改善寡核苷酸的细胞内递送。

Antisense & nucleic acid drug development Pub Date : 2001-02-01 DOI: 10.1089/108729001750072083

Y. Liu, R. Bergan

{"title":"Improved intracellular delivery of oligonucleotides by square wave electroporation.","authors":"Y. Liu, R. Bergan","doi":"10.1089/108729001750072083","DOIUrl":"https://doi.org/10.1089/108729001750072083","url":null,"abstract":"Prior studies have shown that electroporation is a simple and effective method for the introduction of oligonucleotides (ODN) into cells. In ex vivo bone marrow purging models, electroporation of ODN into cells has been associated with selective killing of human neoplastic cells while sparing hematopoietic stem cells. Prior studies used conventional electroporation methods (i.e., exponential decay) to introduce ODN into cells. Square wave electroporation allows the delivery of a more defined and regulated electrical pulse and is associated with high transfection efficiencies in a variety of systems. The current study was undertaken to determine whether square wave electroporation was more effective than exponential decay electroporation for the delivery of ODN into hematopoietic cells. Using fluorescein-tagged ODN and K562, chronic myelogenous leukemia (CML) cells, higher transfection rates were observed after square wave electroporation. In addition, c-myc antisense ODN were more effective in reducing c-myc protein when introduced by square wave electroporation, as compared with introduction by exponential decay electroporation. Square wave electroporation is thus identified as the optimal method for delivering ODN into hematopoietic cells.","PeriodicalId":7996,"journal":{"name":"Antisense & nucleic acid drug development","volume":"26 1","pages":"7-14"},"PeriodicalIF":0.0,"publicationDate":"2001-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"77579033","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 24

Transdermal delivery of antisense oligonucleotides can induce changes in gene expression in vivo. 反义寡核苷酸经皮递送可诱导体内基因表达的变化。

Antisense & nucleic acid drug development Pub Date : 2001-02-01 DOI: 10.1089/108729001750072074

R. Brand, T. L. Hannah, J. Norris, P. Iversen

{"title":"Transdermal delivery of antisense oligonucleotides can induce changes in gene expression in vivo.","authors":"R. Brand, T. L. Hannah, J. Norris, P. Iversen","doi":"10.1089/108729001750072074","DOIUrl":"https://doi.org/10.1089/108729001750072074","url":null,"abstract":"The potential for using antisense compounds as therapeutic agents has generated great enthusiasm. Strategies for delivery of these compounds are, therefore, of great interest. Transdermal iontophoresis has been used successfully as an enhancement technique for the transdermal delivery of these compounds in vitro. The effectiveness of using percutaneous penetration as a means to deliver therapeutic levels of these compounds in vivo, however, remains to be demonstrated. The purpose of this work was to demonstrate the ability of iontophoretically delivered compounds to alter enzyme levels in the intact rat. A C5 propyne-modified phosphorothioate oligonucleotide (PS-ODN) targeted to the cytochrome p450-3A2 (CYP3A2) mRNA translational start site and the reverse sequence, used as a control, were synthesized. A patch containing either an oligonucleotide or a buffer control was placed on the animal's back, and an iontophoretic current of 0.5 mA/cm2 was applied for 3.5 hours. Twenty-four hours later, CYP3A2 levels were measured noninvasively using the midazolam-induced sleeping rat model. Liver and small intestinal microsomes were made after completion of sleep studies and assayed for CYP3A2, CYP1A1/2, CYP2B1/2, and CYP2E1. Midozolam-treated animals with antisense to CYP3A2 slept significantly longer than did the controls (p < 0.05). CYP3A2 levels were significantly lower in liver microsomes from antisense-treated animals than in either buffer control (p < 0.001) or reverse sequence animals (p < 0.05). The reverse sequence was also significantly different from the buffer control (p < 0.01), indicating a nonspecific effect of the PS background. Nontarget cytochrome levels were not altered by treatment. There were no significant differences in small intestine CYP3A2 levels between treatment groups. These data demonstrate that transdermally delivered PS-ODN can reach concentrations sufficient to induce changes in specific target enzymes in vivo. Further studies are warranted to investigate potential uses for these molecules.","PeriodicalId":7996,"journal":{"name":"Antisense & nucleic acid drug development","volume":"155 1","pages":"1-6"},"PeriodicalIF":0.0,"publicationDate":"2001-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"73416128","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 21

Targeting the human mdr1 gene by 125I-labeled triplex-forming oligonucleotides. 通过125i标记的三联体形成寡核苷酸靶向人mdr1基因。

Antisense & nucleic acid drug development Pub Date : 2000-12-01 DOI: 10.1089/OLI.1.2000.10.443

O. Sedelnikova, I. Panyutin, A. N. Luu, M. Reed, T. Licht, M. Gottesman, R. Neumann

{"title":"Targeting the human mdr1 gene by 125I-labeled triplex-forming oligonucleotides.","authors":"O. Sedelnikova, I. Panyutin, A. N. Luu, M. Reed, T. Licht, M. Gottesman, R. Neumann","doi":"10.1089/OLI.1.2000.10.443","DOIUrl":"https://doi.org/10.1089/OLI.1.2000.10.443","url":null,"abstract":"Antigene radiotherapy is our approach to targeting specific sites in the genome by combining the highly localized DNA damage produced by the decay of Auger electron emitters, such as 125I, with the sequence-specific action of triplex-forming oligonucleotides (TFO). As a model, we used the multidrug resistance gene (mdr1) overexpressed and amplified nearly 100 times in the human KB-V1 carcinoma cell line. Phosphodiester pyrrazolopyrimidine dG (PPG)-modified TFO complementary to the polypurine-polypyrimidine region of the mdr1 gene were synthesized and labeled with 125I-dCTP at the C5 position of two cytosines by the primer extension method. 125I-TFO were delivered into KB-V1 cells with several delivery systems. DNA from the 125I-TFO-treated cells was recovered and analyzed for sequence-specific cleavage in the mdr1 target by Southern hybridization. Experiments with plasmid DNA containing the mdr1 polypurine-polypyrimidine region and with purified genomic DNA confirmed the ability of the designed 125I-TFO to bind to and introduce double-strand breaks into the target sequence. We showed that 125I-TFO in nanomolar concentrations can recognize and cleave a target sequence in the mdr1 gene in situ, that is, within isolated nuclei and intact digitonin-permeabilized cells. Our results demonstrate the ability of 125I-TFO to target specific sequences in their natural environment, that is, within the eukaryotic nucleus. The nearly 100-fold amplification of the mdr1 gene in KB-V1 cells affords a very useful cell culture model for evaluation of methods to produce sequence-specific DNA double-strand breaks for gene-specific radiotherapy.","PeriodicalId":7996,"journal":{"name":"Antisense & nucleic acid drug development","volume":"8 1","pages":"443-52"},"PeriodicalIF":0.0,"publicationDate":"2000-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"73147553","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 19

A short phosphodiester window is sufficient to direct RNase H-dependent RNA cleavage by antisense peptide nucleic acid. 一个短的磷酸二酯窗口足以指导RNase h依赖性RNA被反义肽核酸切割。

Antisense & nucleic acid drug development Pub Date : 2000-12-01 DOI: 10.1089/OLI.1.2000.10.463

C. Malchere, J. Verheijen, S. van der Laan, L. Bastide, J. V. van Boom, B. Lebleu, I. Robbins

引用次数: 23