Adenovirus-Mediated Gene Transfer In Vivo to Cerebral Blood Vessels and Perivascular Tissue

Abstract

Gene transfer to blood vessels in vivo generally requires interruption of blood flow. Thus, gene transduction to cerebral blood vessels in vivo has not yet been achieved. In this study, we injected replication-deficient adenovirus into cerebrospinal fluid in an attempt to transduce genes to cerebral blood vessels. Recombinant adenovirus (1 x 10(9) infectious units) expressing nuclear-targeted bacterial beta-galactosidase driven by the cytomegalovirus promoter was injected into the cisterna magna of Sprague-Dawley rats. The brains were examined histochemically after staining with 5-bromo-4-chloro-3-indolyl-beta-D-galactopyranoside 1 to 7 days after injection of adenovirus. Leptomeningeal cells overlying the major arteries were efficiently transduced, and adventitial cells of large vessels and smooth muscle cells of small vessels were occasionally stained. beta-Galactosidase was expressed on days 1 and 3 after injection but was undetectable by day 7. Expression of the gene was 'targeted' by altering the position of the head. When viral suspension was injected while the rat was in a nose-down position, the reporter gene was expressed extensively on the ventral surface of the brain, especially along the circle of Willis. When the position was changed to the nose-up or lateral position, the inferior or lateral region of the brain was stained primarily. Administration of the virus into the lateral ventricle provided extensive expression in ependymal cells and leptomeninges with some transduction to cerebral blood vessels. Thus, adenovirus injected into cerebrospinal fluid provides gene transfer in vivo to cerebral blood vessels and, with greater efficiency, to perivascular tissue. Furthermore, cisternal delivery may target specific brain regions by positioning of the head.(ABSTRACT TRUNCATED AT 250 WORDS)

A direct mechanical method for accurate and efficient adenoviral vector delivery to tissues

“…Experimental studies have demonstrated successful transfer of a variety of recombinant genes in the cardiovascular system both ex vivo and in vivo in animals and humans [1][2][3][4][5][6][7][8][9][10][11][12][13] suggesting that gene transfer technology may, in future, offer an effective therapeutic option for the spectrum of human cardiovascular diseases. [14][15][16][17][18] Most vascular gene transfer studies, to date, have utilized replication-incompetent adenovirus as a delivery vehicle for recombinant cDNA.…”

“…Ex vivo delivery of this vector has been carried out by immersion of arterial tissue in a chamber containing vector-solution, 3,5,11 while in vivo delivery has been carried out through the lumen of an artery, that is, by direct injection, or via its outer ''adventitial'' layer, that is, perivascularly. 2,4,7,8,10,13,[19][20][21] A number of groups have demonstrated the ability of perivascular (adventitial) gene delivery to favorably and profoundly modify vasomotor function in a variety of cardiovascular models [3][4][5][11][12][13]19,20,22 and this approach, which has been reported to minimize the proinflammatory effects of adenoviral vectors, 10 can be used as an effective adjunct to the deployment of intravascular gene delivery stents and catheters. In vivo, current vascular gene delivery techniques have limited ability to produce specific targeting and selective localization of vector in a desired arterial territory or individual vessel.…”

Liposome-mediated transfer of the bcl-2 gene results in neuroprotection after in vivo transient focal cerebral ischemia in an animal model

“…Several studies have shown that gene transfer to the brain by intrathecal injection of vecGene Therapy tors may be efficient and may take part in different compartments. [21][22][23] AV vectors carrying the ␤-galactosidase gene were injected into rat CSF and produced transgene expression in cerebral blood vessels and perivasucular tissue. 21 Meuli-Simmen et al 23 have achieved liposomemediated gene transfer to mouse brain by intrathecal administration.…”

“…[21][22][23] AV vectors carrying the ␤-galactosidase gene were injected into rat CSF and produced transgene expression in cerebral blood vessels and perivasucular tissue. 21 Meuli-Simmen et al 23 have achieved liposomemediated gene transfer to mouse brain by intrathecal administration. Intrathecal injection of lipoplexes containing the chloramphenicol acetyltransferase (CAT) reporter gene showed that CAT activity in the brain reached peak levels between 24 h and 1 week after injection, and was still present as traces 3 weeks after a single injection.…”

Age-related neuronal vulnerability to brain ischemia: A potential target of gene therapy

“…In that case, needs of stopping blood flow may be a critical limitation in the cerebral circulation. Therefore, we have examined efficacy of an alternative method, perivascular approach [22,23]. To deliver the foreign gene to cerebral blood vessels, we administered an adenoviral vectors encoding cDNA of bacterial B-galactosidase as a reporter gene into cerebrospinal fluid (CSF) via the cisterna magna [24].…”