PMID- 27718541 OWN - NLM STAT- MEDLINE DCOM- 20170728 LR - 20181113 IS - 1471-4159 (Electronic) IS - 0022-3042 (Print) IS - 0022-3042 (Linking) VI - 140 IP - 2 DP - 2017 Jan TI - 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. PG - 216-230 LID - 10.1111/jnc.13861 [doi] AB - 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 (AAV9) has been shown to transduce cells in the CNS following intravascular administration in rodents, cats, pigs, and non-human primates. These results suggest that AAV9 is capable of crossing the BBB. However, mechanisms that govern AAV9 transendothelial trafficking at the BBB remain unknown. Furthermore, possibilities that AAV9 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 AAV9 to AAV2, a serotype that does not cross the BBB or transduce endothelial cells effectively in vivo. Results of our in vitro studies indicate that AAV9 penetrates brain microvascular endothelial cells barriers more effectively than AAV2, but has reduced transduction efficiency. In addition, our data suggest that (i) AAV9 penetrates endothelial barriers through an active, cell-mediated process, and (ii) AAV9 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. CI - (c) 2016 International Society for Neurochemistry. FAU - Merkel, Steven F AU - Merkel SF AUID- ORCID: 0000-0003-2889-5187 AD - Department of Pathology and Laboratory Medicine, The Lewis Katz School of Medicine at Temple University, Philadelphia, Pennsylvania, USA. AD - Center for Substance Abuse Research, The Lewis Katz School of Medicine at Temple University, Philadelphia, Pennsylvania, USA. FAU - Andrews, Allison M AU - Andrews AM AUID- ORCID: 0000-0003-1752-7450 AD - Department of Pathology and Laboratory Medicine, The Lewis Katz School of Medicine at Temple University, Philadelphia, Pennsylvania, USA. AD - Center for Substance Abuse Research, The Lewis Katz School of Medicine at Temple University, Philadelphia, Pennsylvania, USA. FAU - Lutton, Evan M AU - Lutton EM AD - Department of Pathology and Laboratory Medicine, The Lewis Katz School of Medicine at Temple University, Philadelphia, Pennsylvania, USA. FAU - Mu, Dakai AU - Mu D AD - Department of Neurology, The Massachusetts General Hospital, Charlestown, Massachusetts, USA. FAU - Hudry, Eloise AU - Hudry E AD - Department of Neurology, The Massachusetts General Hospital, Charlestown, Massachusetts, USA. AD - NeuroDiscovery Center, Harvard Medical School, Boston, Massachusetts, USA. AD - Alzheimer Research Unit, The Massachusetts General Hospital Institute for Neurodegenerative Disease, Charlestown, Massachusetts, USA. FAU - Hyman, Bradley T AU - Hyman BT AD - Department of Neurology, The Massachusetts General Hospital, Charlestown, Massachusetts, USA. AD - NeuroDiscovery Center, Harvard Medical School, Boston, Massachusetts, USA. AD - Alzheimer Research Unit, The Massachusetts General Hospital Institute for Neurodegenerative Disease, Charlestown, Massachusetts, USA. FAU - Maguire, Casey A AU - Maguire CA AD - Department of Neurology, The Massachusetts General Hospital, Charlestown, Massachusetts, USA. AD - NeuroDiscovery Center, Harvard Medical School, Boston, Massachusetts, USA. FAU - Ramirez, Servio H AU - Ramirez SH AD - Department of Pathology and Laboratory Medicine, The Lewis Katz School of Medicine at Temple University, Philadelphia, Pennsylvania, USA. AD - Center for Substance Abuse Research, The Lewis Katz School of Medicine at Temple University, Philadelphia, Pennsylvania, USA. AD - Shriners Hospitals Pediatric Research Center, The Lewis Katz School of Medicine at Temple University, Philadelphia, Pennsylvania, USA. LA - eng GR - K99 AG047336/AG/NIA NIH HHS/United States GR - P30 DA013429/DA/NIDA NIH HHS/United States GR - P30 NS045776/NS/NINDS NIH HHS/United States GR - R01 NS086570/NS/NINDS NIH HHS/United States GR - T32 DA007237/DA/NIDA NIH HHS/United States PT - Comparative Study PT - Journal Article PT - Research Support, N.I.H., Extramural PT - Research Support, Non-U.S. Gov't DEP - 20161215 PL - England TA - J Neurochem JT - Journal of neurochemistry JID - 2985190R SB - IM MH - Blood-Brain Barrier/cytology/*virology MH - Brain/metabolism/*virology MH - Cell Movement/*physiology MH - Cells, Cultured MH - *Dependovirus MH - Endothelial Cells/*virology MH - Humans MH - Tight Junctions/*virology MH - Transcytosis/physiology PMC - PMC5298820 MID - NIHMS822576 OTO - NOTNLM OT - adeno-associated virus OT - blood-brain barrier OT - gene therapy OT - neurological disorders COIS- The authors declare no competing financial interests. EDAT- 2016/10/09 06:00 MHDA- 2017/07/29 06:00 PMCR- 2018/01/01 CRDT- 2016/10/09 06:00 PHST- 2016/03/29 00:00 [received] PHST- 2016/08/07 00:00 [revised] PHST- 2016/09/19 00:00 [accepted] PHST- 2016/10/09 06:00 [pubmed] PHST- 2017/07/29 06:00 [medline] PHST- 2016/10/09 06:00 [entrez] PHST- 2018/01/01 00:00 [pmc-release] AID - 10.1111/jnc.13861 [doi] PST - ppublish SO - J Neurochem. 2017 Jan;140(2):216-230. doi: 10.1111/jnc.13861. Epub 2016 Dec 15.