Antisense preproendothelin-oligoDNA therapy for vasospasm in a canine model of subarachnoid hemorrhage

Ohkuma, H Parney, I Megyesi, J Ghahary, A Findlay, JM

H. Ohkuma et al., Antisense preproendothelin-oligoDNA therapy for vasospasm in a canine model of subarachnoid hemorrhage, J NEUROSURG, 90(6), 1999, pp. 1105-1114

43

INGLESE

Article

Neurology,"Neurosciences & Behavoir

JOURNAL OF NEUROSURGERY

0022-3085 → ACNP

90

6

1999

1105 - 1114

ISI

0022-3085(199906)90:6<1105:APTFVI>2.0.ZU;2-S

Object. The purpose of this study is twofold: 1) to test antisense genetic techniques used in the prevention of cerebral vasospasm in a canine model o f subarachnoid hemorrhage (SAH), targeting the endothelin-1 (ET-I) gene; an d 2) to determine if fibrinolysis of subarachnoid clot with recombinant tis sue plasminogen activator (rtPA) could enhance the effect of antisense trea tment. Methods. A total of 39 dogs were studied in this experiment. Placebo (six a nimals), rtPA (six animals), antisense preproET-1 oligodeoxynucleotide (ASO D; five animals), or rtPA plus ASOD (combined treatment; six animals) was i njected into the cisterna magna 30 minutes after a second SAH was induced o n the 2nd day of the experiment. The animals were observed until Day 7, whe n they underwent follow-up angiography and then were killed; their basilar arteries were removed for analysis. Control animals included in this study (two animals in each group) received placebo, rtPA, ASOD, or rtPA plus ASOD without induction of SAH, or rtPA with mismatched (nonsense) preproET-1 ol igodeoxynucleotide following SAH. Six additional dogs were analyzed earlier following SAH. Dogs that received placebo developed severe vasospasm (51 +/- 8% of baselin e caliber). Administration of ASOD alone resulted in a mild reduction in va sospasm (64 +/- 13% of baseline caliber) and rtPA alone resulted in a moder ate reduction in vasospasm (81 +/- 5% of baseline caliber); however, the co mbined therapy of rtPA plus ASOD almost completely prevented vasospasm (95 +/- 6% of baseline caliber), which was significantly different from all oth er groups (p < 0.05). Morphological analysis of the basilar arteries yielde d results similar to angiography with respect to vasospasm severity. The AS OD treatment combined with rtPA resulted in reduced ET-1 expression, as dem onstrated by immunohistochemical staining of the arteries, and reduced prep roET-1 levels on Day 4, as measured by reverse transcription-polymerase cha in reaction. Nonsense DNA sequences had no effect on the vessels. Conclusions. Antisense preproET-1 oligodeoxynucleotide treatment, when comb ined with clot lysis caused by rtPA, reduced vasospasm in the canine model of SAH, and this effect appeared to be related to reduced ET-1 synthesis. T he results of this experiment support a causative role for ET-1 early in th e course of vasospasm development in dogs. The apparent additive therapeuti c effects of antisense and fibrinolytic treatments could be due to clot lys is, which allows better delivery of oligodeoxynucleotides to arteries withi n the subarachnoid space.