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Trafficking of adeno‐associated virus vectors across a model of the blood–brain barrier; a comparative study of transcytosis and transduction using primary human brain endothelial cells
Author(s) -
Merkel Steven F.,
Andrews Allison M.,
Lutton Evan M.,
Mu Dakai,
Hudry Eloise,
Hyman Bradley T.,
Maguire Casey A.,
Ramirez Servio H.
Publication year - 2017
Publication title -
journal of neurochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.75
H-Index - 229
eISSN - 1471-4159
pISSN - 0022-3042
DOI - 10.1111/jnc.13861
Subject(s) - transcytosis , blood–brain barrier , transduction (biophysics) , microbiology and biotechnology , biology , endothelial stem cell , human brain , adeno associated virus , central nervous system , tight junction , neuroscience , in vitro , immunology , cell , endocytosis , vector (molecular biology) , recombinant dna , biophysics , biochemistry , gene
Developing therapies for central nervous system ( CNS ) diseases is exceedingly difficult because of the blood–brain barrier ( BBB ). Notably, emerging technologies may provide promising new options for the treatment of CNS disorders. Adeno‐associated virus serotype 9 ( AAV 9) has been shown to transduce cells in the CNS following intravascular administration in rodents, cats, pigs, and non‐human primates. These results suggest that AAV 9 is capable of crossing the BBB . However, mechanisms that govern AAV 9 transendothelial trafficking at the BBB remain unknown. Furthermore, possibilities that AAV 9 may transduce brain endothelial cells or affect BBB integrity still require investigation. Using primary human brain microvascular endothelial cells as a model of the human BBB , we performed transduction and transendothelial trafficking assays comparing AAV 9 to AAV 2, a serotype that does not cross the BBB or transduce endothelial cells effectively in vivo . Results of our in vitro studies indicate that AAV 9 penetrates brain microvascular endothelial cells barriers more effectively than AAV 2, but has reduced transduction efficiency. In addition, our data suggest that (i) AAV 9 penetrates endothelial barriers through an active, cell‐mediated process, and (ii) AAV 9 fails to disrupt indicators of BBB integrity such as transendothelial electrical resistance, tight junction protein expression/localization, and inflammatory activation status. Overall, this report shows how human brain endothelial cells configured in BBB models can be utilized for evaluating transendothelial movement and transduction kinetics of various AAV capsids. Importantly, the use of a human in vitro BBB model can provide import insight into the possible effects that candidate AVV gene therapy vectors may have on the status of BBB integrity.Read the Editorial Highlight for this article on page 192.