Distribution and stability of antisense phosphorothioate oligonucleotides in rodent brain following direct intraparenchymal controlled-rate infusion

Broaddus, William C.; Prabhu, Sujit S.; Gillies, G. T.; Neal, Jeffrey; Conrad, William S.; Chen, Zhijian; Fillmore, Helen; Young, Harold F.

doi:10.3171/jns.1998.88.4.0734

Cited by 51 publications

(19 citation statements)

References 29 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…To achieve uniform and reproducible results, we employed a slow and rate-controlled positive pressure pump that we have used previously to deliver antisense oligonucleotides into the brain of Fischer 344 rats. 23 We found that 5 ϫ 10 9 PFU of Ad␤gal in a volume of 150 L and infused at a rate of 1 L/minute was the most efficient condition for virus delivery compared with the same dose of virus in either 50 or 3 ϫ 50 L, as determined by ␤-gal staining of tumor sections (Fig 6). Using this procedure, we were able to achieve 40% infection of the tumor, whereas 50 and 3 ϫ 50 L produced 5% and 25% infection, respectively.…”

Section: Rat Rt2 Cells Express Low Levels Of CDmentioning

confidence: 77%

Radiosensitization of rat glioma with bromodeoxycytidine and adenovirus expressing herpes simplex virus-thymidine kinase delivered by slow, rate-controlled positive pressure infusion

et al. 2000

View full text Add to dashboard Cite

Infection of rat RT2 glioma cells in vitro with an adenovirus (ADV-TK) expressing herpes simplex virus (HSV) thymidine kinase (TK) and subsequent exposure to 5-bromo-2Ј-deoxycytidine (BrdC), which is specifically incorporated into ADV-TK-infected cell DNA as 5-bromo-2Ј-deoxyuridine (BrdU), results in significant radiosensitization (sensitizer enhancement ratio: 1.4 -2.3) compared with Ad␤gal-infected cells. Cell killing correlated well with increased BrdU DNA incorporation and with apoptosis. Whereas radiation (4 Gy) alone was relatively ineffective in inducing apoptosis, treatment with HSV-TK/BrdC resulted in BrdC dose-(10 -100 M) and time-dependent (24 -48 hours) increases, and the combination of the two treatments produced a synergistic response (1.5-to 2-fold). To investigate the effects of the ADV-TK/BrdC treatment in vivo, RT2 cells were grown as soft tissue tumors in Fischer 344 rats and conditions for virus infusion were optimized by altering the volume and rate of infusion using a rate-controlled positive pressure device. We found that relatively large volumes (100 -150 L) of virus delivered at rates of Յ1 L/minute were optimal and gave uniform and reproducible results. Using these optimal infusion conditions, we were able to achieve 40% adenovirus infection in the tumor. Infection of RT2 tumors with ADV-TK and continuous administration of BrdC from an osmotic pump resulted in significant (.001 Ͻ P Ͻ .009) tumor regression 6 days after radiation (30 Gy delivered as 2 ϫ 5 Gy over 3 days) compared with controls. In situ staining of sectioned tumors with anti-BrdU antibody or by high-performance liquid chromatography analysis of extracted and hydrolyzed tumor DNA confirmed that we obtained efficient and specific incorporation of BrdU into tumor cells. These results suggest that adenovirus-mediated delivery of HSV-TK in combination with BrdC and radiation can potentially be an efficient combination modality for the treatment of gliomas. Cancer Gene Therapy (2000) 7, 778 -788 Key words: Cancer gene therapy; drug delivery; halogenated pyrimidines; osmotic pump; radiation.T he halogenated pyrimidines (HPs) 5-bromo-2Ј-deoxyuridine (BrdU) and 5-iodo-2Ј-deoxyuridine (IdU) have been studied extensively as radiosensitizers of various mammalian cells in culture. [1][2][3] Preclinical and clinical studies have revealed that both BrdU and IdU are relatively efficient radiosensitizers if incorporated at sufficient levels into DNA. 4,5 However, although some clinical success has been achieved with head and neck tumors, attempts to radiosensitize gliomas with HPs have been disappointing, perhaps due to poor incorporation of the halogenated nucleosides into the DNA of these types of tumor cells. 6 Therefore, improved or alternative approaches are needed to treat this disease. The thymidine kinase (tk) gene of herpes simplex virus (HSV) has been used extensively to sensitize cancer cells to the HSV-TK-specific drug ganciclovir (GCV), and quite promising results with glioma have been obtained using various animal tumor mode...

show abstract

Section: Rat Rt2 Cells Express Low Levels Of CDmentioning

confidence: 77%

Radiosensitization of rat glioma with bromodeoxycytidine and adenovirus expressing herpes simplex virus-thymidine kinase delivered by slow, rate-controlled positive pressure infusion

et al. 2000

View full text Add to dashboard Cite

show abstract

“…Antisense oligonucleotide and the drug directly injected into brain with high-flow microinfusion can be safely delivered at high concentration to wide areas of rat brain, bypassing the blood-brain barrier and avoiding the implications after systemic administration (24,25). Clinical trials of this drug in humans have been discouraging (26,27).…”

Section: Discussionmentioning

confidence: 99%

c-Met antisense oligodeoxynucleotides increase sensitivity of human glioma cells to paclitaxel

Chu¹

2010

Oncol Rep

View full text Add to dashboard Cite

Abstract. Cell culture, tissue chemistry and flow cytometry were used to determine whether antisense c-Met oligodeoxynucleotides enhanced the sensitivity of human glioma cells to paclitaxel. A combination of paclitaxel with antisense c-Met oligodeoxynucleotides inhibited cell growth, induced apoptosis and induced c-Met protein expression in U251 and SHG44 human glioma cells more significantly than either paclitaxel or the oligodeoxynucleotides on their own (P<0.01). Thus, c-Met antisense oligodeoxynucleotides increase the sensitivity of human glioma cells to paclitaxel. Combined use of the two agents could be a novel and attractive strategy in human glioma treatment.

show abstract

“…Unmodified phosphodiester oligodeoxynucleotides remain intact after incubation in CSF ex vivo for up to 24 h (Whitesell et al 1993). Accordingly, unmodified oligodeoxynucleotides injected intracerebrally were shown to be rapidly degraded and to disappear within 24 h from the injection site, whilst phosphorothioate oligodeoxynucleotides remained stable and could be detected after 48 h post injection (Szklarczyk & Kaczmarek, 1995;Broaddus et al 1998). The tissue penetration of phosphorothioate AS ODNs infused either intrastriatally or i.c.v.…”

Section: Experimental Controlsmentioning

confidence: 99%

“…injections is the use of osmotic minipumps, enabling continuous infusion for several days Broaddus et al 1999). Intraparenchymal controlled-rate infusion has also been employed (Broaddus et al 1998). Examples of doses of AS ODNs and modes of intracerebral delivery are provided in recent reviews Weiss et al 1997;Phillips, 1997).…”

Section: Experimental Controlsmentioning

confidence: 99%

Antisense Oligonucleotides in the Study of Central Mechanisms of the Cardiovascular Regulation

Čulman¹

2000

Experimental Physiology

View full text Add to dashboard Cite

In the past several years, the progress in the development of molecular biology and the elucidation of gene sequences has created new approaches for studying biological functions and developing new diagnostic and therapeutic strategies. One of the most exciting advances has been the development of antisense technology, which represents a new strategy allowing modulation of protein synthesis with high specificity by preventing protein expression at the level of RNA or DNA. Many classical pharmacological approaches in neurobiological research are often based on the inhibition of biologically active proteins, such as receptors for neurotransmitters, or enzymes involved in neurotransmitter synthesis or degradation. The use of antisense oligodeoxynucleotides offers an alternative tool to manipulate selectively the expression of neurotransmitters or their receptors in neuronal tissue. This approach is especially useful when selective, high‐affinity antagonists are not available. As a result, this technology has gained acceptance in the study of cell signalling mechanisms and the molecular basis of neuronal function. This paper provides a brief background to the antisense technique and explores methodological aspects, particularly in the whole animal. The use of the antisense technology in studies focused on central mechanisms regulating the cardiovascular system is then discussed.

show abstract

Distribution and stability of antisense phosphorothioate oligonucleotides in rodent brain following direct intraparenchymal controlled-rate infusion

Abstract: The authors found that PS-ODNs can be safely delivered in high concentrations to wide areas of rat brain by using high-flow microinfusion and are stable even after 48 hours in situ.

Cited by 51 publications

References 29 publications

Radiosensitization of rat glioma with bromodeoxycytidine and adenovirus expressing herpes simplex virus-thymidine kinase delivered by slow, rate-controlled positive pressure infusion

Radiosensitization of rat glioma with bromodeoxycytidine and adenovirus expressing herpes simplex virus-thymidine kinase delivered by slow, rate-controlled positive pressure infusion

c-Met antisense oligodeoxynucleotides increase sensitivity of human glioma cells to paclitaxel

Antisense Oligonucleotides in the Study of Central Mechanisms of the Cardiovascular Regulation

Contact Info

Product

Resources

About