Gp340 and syndecan inhibition based microbicide for HIV

描述（由申请人提供）：在缺乏保护性疫苗的情况下，教育和对艾滋病毒有活性的杀微生物剂是控制全球流行病的最佳方法。我们的研究项目研究生殖道传播的最早期事件。我们已经确定了一种由生殖道上皮细胞表达的蛋白质，可以作为抑制HIV传播的潜在靶点，称为gp-340。我们已经证明，gp-340表达在阴道和宫颈上皮细胞的细胞表面上，在体内，体外和离体和结合HIV包膜。对生殖道传播具有重要意义的是，病毒的gp-340结合导致病毒的感染性和半衰期增加。由生殖道组织和细胞系表达的GP-340也介导HIV的转胞吞作用，即大分子从细胞的一侧到另一侧的囊泡运输。第二种称为多配体蛋白聚糖的分子已被研究，并显示出具有类似的跨感染和转胞吞特性，也由生殖道细胞表达。我们已经确定了一种肽抑制剂的包膜结合糖蛋白-340，阻止糖蛋白-340介导的转感染和转胞吞作用在体外和体外模型的生殖道传播。该肽含有抑制多配体蛋白聚糖介导的转感染的基序的一部分，我们将修饰该肽以抑制与猕猴gp 340和多配体蛋白聚糖的包膜结合，并将其开发成杀微生物剂。gp-340和多配体蛋白聚糖在递送至生殖道腔后但在与靶细胞相互作用和感染靶细胞之前的感染阶段起作用的这种潜在作用在杀微生物剂设计中是非常有吸引力和新颖的。我们假设，干扰这一过程将抑制或阻断生殖道传播。在本提案的初始R21部分，我们将建立生殖道传播的体外猕猴系统。如果我们证明猕猴gp 340和多配体蛋白聚糖介导转感染和转胞吞作用，并且V3环衍生的肽或改进的形式阻断猕猴gp 340和多配体蛋白聚糖介导的转感染和转胞吞作用，我们将继续进行授权的R33部分。其具体目的是：通过体外试验开发杀微生物剂，并在恒河猴阴道传播模型中测试阻断gp 340和syndecan-HIV Env相互作用对生殖道SIV传播的影响。通过这些具体的目标，我们将开发一种新型的杀微生物剂，并确定生殖道gp-340和syndecan在HIV传播中的作用。如果成功，这些研究将提供一种基于宿主细胞与HIV相互作用的新的杀微生物剂，促进生殖道传播到临床前试验研究。

DESCRIPTION (provided by applicant): Education and microbicides active against HIV represent the best approaches to controlling the epidemic worldwide in the absence of a protective vaccine. Our research program studies the earliest events in genital tract transmission. We have identified a protein expressed by genital tract epithelial cells that could serve as a potential target for inhibition of transmission of HIV called gp-340. We have demonstrated that gp-340 is expressed on the cell surface of vaginal and cervical epithelial cells, in vivo, in vitro, and ex vivo and binds HIV envelope. Of significance to genital tract transmission, gp-340 binding of virus leads to an increase in both the infectivity and half-life of the virus. Gp-340 expressed by genital tract tissue and cell lines also mediates transcytosis of HIV, the vesicular transport of macromolecules from one side of a cell to the other. A second molecule called syndecan has been studied and shown to have similar trans-infection and transcytosis properties and is also expressed by genital tract cells. We have identified a peptide inhibitor of envelope binding to gp-340 that blocks both gp-340 mediated trans-infection and transcytosis in in vitro and ex vivo models of genital tract transmission. This peptide contains a portion of a motif that inhibits syndecan mediated transinfection, as well, and we will modify this peptide to inhibit envelope binding to both macaque gp340 and syndecan and develop it into a microbicide. This potential role of gp-340 and syndecan to act at a stage of infection after delivery to the lumen of the genital tract but prior to interaction with and infection of target cells is very attractive and novel in microbicide design. We hypothesize that interfering with this process will inhibit or block genital tract transmission. In the initial R21 portion of this proposal, we will establish in vitro macaque systems of genital tract transmission. If we demonstrate that macaque gp340 and syndecan mediate trans- infection and transcytosis and V3 loop derived peptides or improved versions block macaque gp340 and syndecan mediated transinfection and transcytosis, we will proceed with the R33 portion of the grant. The specific aims of this are: microbicide development with in vitro testing and to test the effect of blocking gp340 and syndecan-HIV Env interaction on genital tract SIV transmission in the rhesus macaque vaginal transmission model. Through these specific aims, we will develop a new type of microbicide and determine the role of genital tract gp-340 and syndecan in HIV transmission. If successful, these studies will deliver a new microbicide based on host cell interactions with HIV that promote genital tract transmission to preclinical trial studies.