A Novel Adeno-Associated Viral Variant for Efficient and Selective Intravitreal Transduction of Rat Muller Cells

被引:145
作者
Klimczak, Ryan R.
Koerber, James T.
Dalkara, Deniz
Flannery, John G.
Schaffer, David V.
机构
[1] Department of Molecular and Cellular Biology, University of California, Berkeley, CA
[2] Department of Chemical Engineering, Department of Bioengineering, The University of California, Berkeley, CA
[3] Helen Wills Neuroscience Institute, University of California, Berkeley, CA
来源
PLOS ONE | 2009年 / 4卷 / 10期
关键词
RETINAL-DETACHMENT; VISUAL FUNCTION; GENE-TRANSFER; VIRUS; VECTOR; THERAPY; ASTROCYTES; TROPISM; BINDING; VISION;
D O I
10.1371/journal.pone.0007467
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
Background: The pathologies of numerous retinal degenerative diseases can be attributed to a multitude of genetic factors, and individualized treatment options for afflicted patients are limited and cost-inefficient. In light of the shared neurodegenerative phenotype among these disorders, a safe and broad-based neuroprotective approach would be desirable to overcome these obstacles. As a result, gene delivery of secretable-neuroprotective factors to Muller cells, a type of retinal glia that contacts all classes of retinal neurons, represents an ideal approach to mediate protection of the entire retina through a simple and innocuous intraocular, or intravitreal, injection of an efficient vehicle such as an adeno-associated viral vector (AAV). Although several naturally occurring AAV variants have been isolated with a variety of tropisms, or cellular specificities, these vectors inefficiently infect Muller cells via intravitreal injection. Methodology/Principal Findings: We have previously applied directed evolution to create several novel AAV variants capable of efficient infection of both rat and human astrocytes through iterative selection of a panel of highly diverse AAV libraries. Here, in vivo and in vitro characterization of these isolated variants identifies a previously unreported AAV variant ShH10, closely related to AAV serotype 6 (AAV6), capable of efficient, selective Muller cell infection through intravitreal injection. Importantly, this new variant shows significantly improved transduction relative to AAV2 (>60%) and AAV6. Conclusions/Significance: Our findings demonstrate that AAV is a highly versatile vector capable of powerful shifts in tropism from minor sequence changes. This isolated variant represents a new therapeutic vector to treat retinal degenerative diseases through secretion of neuroprotective factors from Muller cells as well as provides new opportunities to study their biological functions in the retina.
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页数:10
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