HIV-envelope-specific DARPin-based microbicide strategies

描述（由申请人提供）：基于hiv包膜特异性darpin的杀微生物剂策略为了成为可行的候选杀微生物剂，抗hiv策略必须高效，阻断相关体外模型系统中的无细胞感染和细胞相关感染，在不干扰免疫功能或引起任何毒性反应的情况下预防体内感染，并且能够以低成本生产。通过最近建立的合作，我们确定了一种新的，高度特异性的方法来抑制HIV感染，使用设计锚蛋白重复蛋白（DARPin）技术。最初的概念证明研究记录了CD4特异性darpin的选择，这些darpin具有高度特异性，并且在纳摩尔水平上有效地预防各种HIV分离株和SIV的体外感染，而不会干扰基本细胞功能。darpin具有独特的特性，表现出卓越的稳定性和特异性，并且生产成本低。将这些特征与预防HIV感染的能力相结合，DARPins代表了一种有希望的预防HIV传播的新策略。在这项新的合作研究的基础上，我们建议选择能够预防粘膜HIV感染的高活性HIV env特异性darpin。我们假设，识别gp120和gp41的env特异性DARPins的鸡尾酒将(i)有效地防止体外无细胞感染和细胞相关感染许多HIV和SIV分离物，在低纳摩尔范围内起作用，（ii）不损害先天或适应性免疫功能，（iii）防止SHIV162P3在猕猴中的粘膜感染，而对局部免疫功能，菌群，这种创新的抗艾滋病毒技术正在Trkola博士的实验室（与Pl¿ckthun博士的小组合作）开发，一旦通过严格的筛选分析被选中，有希望的候选人将在各种体外病毒和免疫系统中进行彻底的测试，使用两位博士建立的相关细胞和组织为基础的分析。Robbiani和Trkola的实验室。然后通过Robbiani博士的实验室（与Blanchard博士合作）在猕猴身上测试主要候选药物的体内活性。利用一种新技术来开发抗HIV Env特异性策略，这种高风险的DARPins方法应该在预防HIV感染和传播方面取得重大进展。

DESCRIPTION (provided by applicant): HIV-envelope-specific DARPin-based microbicide strategies In order to be a viable candidate microbicide, an anti-HIV strategy must be highly effective, block cell-free and cell-associated infections in relevant in vitro model systems, prevent in vivo infection without perturbing immune function or causing any toxic reactions, and be able to be produced at low costs. Through a recently established collaboration, we identified a novel, highly specific approach to inhibit HIV infection using the Designed Ankyrin Repeat Protein (DARPin) technology. The initial proof of concept studies documented the selection of CD4- specific DARPins that are highly specific and efficiently prevent in vitro infection with a wide range of HIV isolates as well as SIV at nanomolar levels, without interfering with basic cellular functions. DARPins possess unique characteristics exhibiting exceptional stability and specificity, and cost little to produce. Combining these features with the ability to prevent HIV infection, DARPins represent a promising novel strategy to explore for the prevention of HIV spread. Expanding on this new collaborative research, we propose to select highly active HIV Env-specific DARPins that will be able to prevent mucosal HIV infection. We hypothesize that cocktails of Env-specific DARPins, which recognize gp120 and gp41, will (i) efficiently prevent in vitro cell-free and cell-associated infection with numerous HIV and SIV isolates, functioning in the low nanomolar range, (ii) not impair innate or adaptive immune functions, and (iii) prevent mucosal infection with SHIV162P3 in macaques, with no adverse impact on local immune function, flora, and pH. This innovative anti-HIV technology is being developed in Dr. Trkola's laboratory (in collaboration with Dr. Pl¿ckthun's group) and, once selected through rigorous screening assays, promising candidates will be thoroughly tested in a variety of in vitro viral and immune systems using relevant cell and tissue-based assays established in both Drs. Robbiani's and Trkola's laboratories. The lead novel candidate(s) will then be tested for in vivo activity in macaques through Dr. Robbiani's laboratory (in collaboration with Dr. Blanchard). Utilizing a novel technology to develop an anti-HIV Env specific strategy, this high-risk DARPins approach should yield critical advances towards preventing HIV infection and spread. Project Summary-Relevance: These studies will provide the first data on the efficacy of this novel topical microbicide to prevent vaginal infection in vivo. With proof of concept that this represents a promising strategy to tackle HIV at the mucosa, we will be able to explore application of this approach to coitally independent modalities, as well as for the potential application against rectal transmission. These data will advance the design and future testing of this novel technology to improve microbicide strategies to test in women and men.