Adenovirus-mediated gene transfer to ischemic brain ischemic flow threshold for transgene expression

Hiroaki Ooboshi, Setsuro Ibayashi, Junichi Takada, Hiroshi Yao, Takanari Kitazono, Masatoshi Fujishima

Research output: Contribution to journalArticle

22 Citations (Scopus)

Abstract

Background and Purpose - Gene therapy may be a promising approach for treatment of brain ischemia, although protein synthesis is generally inhibited in ischemic conditions. Our goal in this study was to examine effects of brain ischemia on transgene expression of adenovirus-mediated gene transfer to ischemic brain. Methods - Brain ischemia was produced by photochemical occlusion of the distal middle cerebral artery of spontaneously hypertensive rats (n=15). Ninety minutes after ischemia, adenoviral vectors encoding bacterial β-galactosidase were injected into ipsilateral (nonischemic [I-n], peri-ischemic [I-p], and ischemic core [I-c] areas) and contralateral parietal (C) cortices. Cerebral blood flow before and during ischemia at each injected area was measured by laser-Doppler flowmetry. Expression of transgene was detected by histochemistry for semiquantitative scoring or by biochemical assay for quantitative analysis. Results - Blood flow to the cortex decreased to 72 ± 10% (mean ± SEM) at I-n, 41 ± 6% at I-p, and 23 ± 3% at I-c after 10 minutes of ischemia. Expression of the reporter gene was consistently detected at C and I-n at each survival period. The semiquantitative score for transgene expression decreased according to severity of ischemia (C, 2.3; I-n, 2.6; I-p, 1.1; I-c, 0.3; mean values). β-Galactosidase activity detected by chemiluminescent assay revealed that the values (mean ± SEM) in the ischemic area (I-p, 15.9 ± 9.2 mU/mg protein; I-c, 1.3 ± 0.5) were significantly smaller than that of the nonischemic area (C, 45.4 ± 6.9). Analysis of cerebral blood flow at I-p revealed that cerebral blood flow threshold for transgene expression was approximately 40% of the resting value. Conclusions - Adenovirus-mediated gene transfer into the ischemic brain provided effective expression of transgene at the nonischemic and peri-ischemic areas. Gene transfer to the ischemic brain may be a promising approach for treatment of ischemic penumbra.

Original languageEnglish
Pages (from-to)1043-1047
Number of pages5
JournalStroke
Volume32
Issue number4
DOIs
Publication statusPublished - Jan 1 2001

Fingerprint

Cerebrovascular Circulation
Transgenes
Adenoviridae
Ischemia
Brain Ischemia
Galactosidases
Brain
Genes
Luminescent Measurements
Parietal Lobe
Laser-Doppler Flowmetry
Middle Cerebral Artery Infarction
Inbred SHR Rats
Reporter Genes
Genetic Therapy
Proteins
Therapeutics

All Science Journal Classification (ASJC) codes

  • Clinical Neurology
  • Cardiology and Cardiovascular Medicine
  • Advanced and Specialised Nursing

Cite this

Adenovirus-mediated gene transfer to ischemic brain ischemic flow threshold for transgene expression. / Ooboshi, Hiroaki; Ibayashi, Setsuro; Takada, Junichi; Yao, Hiroshi; Kitazono, Takanari; Fujishima, Masatoshi.

In: Stroke, Vol. 32, No. 4, 01.01.2001, p. 1043-1047.

Research output: Contribution to journalArticle

Ooboshi, Hiroaki ; Ibayashi, Setsuro ; Takada, Junichi ; Yao, Hiroshi ; Kitazono, Takanari ; Fujishima, Masatoshi. / Adenovirus-mediated gene transfer to ischemic brain ischemic flow threshold for transgene expression. In: Stroke. 2001 ; Vol. 32, No. 4. pp. 1043-1047.
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abstract = "Background and Purpose - Gene therapy may be a promising approach for treatment of brain ischemia, although protein synthesis is generally inhibited in ischemic conditions. Our goal in this study was to examine effects of brain ischemia on transgene expression of adenovirus-mediated gene transfer to ischemic brain. Methods - Brain ischemia was produced by photochemical occlusion of the distal middle cerebral artery of spontaneously hypertensive rats (n=15). Ninety minutes after ischemia, adenoviral vectors encoding bacterial β-galactosidase were injected into ipsilateral (nonischemic [I-n], peri-ischemic [I-p], and ischemic core [I-c] areas) and contralateral parietal (C) cortices. Cerebral blood flow before and during ischemia at each injected area was measured by laser-Doppler flowmetry. Expression of transgene was detected by histochemistry for semiquantitative scoring or by biochemical assay for quantitative analysis. Results - Blood flow to the cortex decreased to 72 ± 10{\%} (mean ± SEM) at I-n, 41 ± 6{\%} at I-p, and 23 ± 3{\%} at I-c after 10 minutes of ischemia. Expression of the reporter gene was consistently detected at C and I-n at each survival period. The semiquantitative score for transgene expression decreased according to severity of ischemia (C, 2.3; I-n, 2.6; I-p, 1.1; I-c, 0.3; mean values). β-Galactosidase activity detected by chemiluminescent assay revealed that the values (mean ± SEM) in the ischemic area (I-p, 15.9 ± 9.2 mU/mg protein; I-c, 1.3 ± 0.5) were significantly smaller than that of the nonischemic area (C, 45.4 ± 6.9). Analysis of cerebral blood flow at I-p revealed that cerebral blood flow threshold for transgene expression was approximately 40{\%} of the resting value. Conclusions - Adenovirus-mediated gene transfer into the ischemic brain provided effective expression of transgene at the nonischemic and peri-ischemic areas. Gene transfer to the ischemic brain may be a promising approach for treatment of ischemic penumbra.",
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AU - Ooboshi, Hiroaki

AU - Ibayashi, Setsuro

AU - Takada, Junichi

AU - Yao, Hiroshi

AU - Kitazono, Takanari

AU - Fujishima, Masatoshi

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N2 - Background and Purpose - Gene therapy may be a promising approach for treatment of brain ischemia, although protein synthesis is generally inhibited in ischemic conditions. Our goal in this study was to examine effects of brain ischemia on transgene expression of adenovirus-mediated gene transfer to ischemic brain. Methods - Brain ischemia was produced by photochemical occlusion of the distal middle cerebral artery of spontaneously hypertensive rats (n=15). Ninety minutes after ischemia, adenoviral vectors encoding bacterial β-galactosidase were injected into ipsilateral (nonischemic [I-n], peri-ischemic [I-p], and ischemic core [I-c] areas) and contralateral parietal (C) cortices. Cerebral blood flow before and during ischemia at each injected area was measured by laser-Doppler flowmetry. Expression of transgene was detected by histochemistry for semiquantitative scoring or by biochemical assay for quantitative analysis. Results - Blood flow to the cortex decreased to 72 ± 10% (mean ± SEM) at I-n, 41 ± 6% at I-p, and 23 ± 3% at I-c after 10 minutes of ischemia. Expression of the reporter gene was consistently detected at C and I-n at each survival period. The semiquantitative score for transgene expression decreased according to severity of ischemia (C, 2.3; I-n, 2.6; I-p, 1.1; I-c, 0.3; mean values). β-Galactosidase activity detected by chemiluminescent assay revealed that the values (mean ± SEM) in the ischemic area (I-p, 15.9 ± 9.2 mU/mg protein; I-c, 1.3 ± 0.5) were significantly smaller than that of the nonischemic area (C, 45.4 ± 6.9). Analysis of cerebral blood flow at I-p revealed that cerebral blood flow threshold for transgene expression was approximately 40% of the resting value. Conclusions - Adenovirus-mediated gene transfer into the ischemic brain provided effective expression of transgene at the nonischemic and peri-ischemic areas. Gene transfer to the ischemic brain may be a promising approach for treatment of ischemic penumbra.

AB - Background and Purpose - Gene therapy may be a promising approach for treatment of brain ischemia, although protein synthesis is generally inhibited in ischemic conditions. Our goal in this study was to examine effects of brain ischemia on transgene expression of adenovirus-mediated gene transfer to ischemic brain. Methods - Brain ischemia was produced by photochemical occlusion of the distal middle cerebral artery of spontaneously hypertensive rats (n=15). Ninety minutes after ischemia, adenoviral vectors encoding bacterial β-galactosidase were injected into ipsilateral (nonischemic [I-n], peri-ischemic [I-p], and ischemic core [I-c] areas) and contralateral parietal (C) cortices. Cerebral blood flow before and during ischemia at each injected area was measured by laser-Doppler flowmetry. Expression of transgene was detected by histochemistry for semiquantitative scoring or by biochemical assay for quantitative analysis. Results - Blood flow to the cortex decreased to 72 ± 10% (mean ± SEM) at I-n, 41 ± 6% at I-p, and 23 ± 3% at I-c after 10 minutes of ischemia. Expression of the reporter gene was consistently detected at C and I-n at each survival period. The semiquantitative score for transgene expression decreased according to severity of ischemia (C, 2.3; I-n, 2.6; I-p, 1.1; I-c, 0.3; mean values). β-Galactosidase activity detected by chemiluminescent assay revealed that the values (mean ± SEM) in the ischemic area (I-p, 15.9 ± 9.2 mU/mg protein; I-c, 1.3 ± 0.5) were significantly smaller than that of the nonischemic area (C, 45.4 ± 6.9). Analysis of cerebral blood flow at I-p revealed that cerebral blood flow threshold for transgene expression was approximately 40% of the resting value. Conclusions - Adenovirus-mediated gene transfer into the ischemic brain provided effective expression of transgene at the nonischemic and peri-ischemic areas. Gene transfer to the ischemic brain may be a promising approach for treatment of ischemic penumbra.

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