Development of novel quantitative methods for in vivo assessment of mucosal irrit

描述（由申请人提供）：本项目的目标是开发敏感和定量成像和细胞方法，用于评价阴道和直肠粘膜对旨在预防HIV感染的非疫苗生物医学预防（nBP）产品的反应。安全有效的nBP策略有助于降低艾滋病毒的发病率，挽救数百万人的生命。虽然近年来出现了几种有希望的候选nBP，并进展到临床试验，但令人失望的结果是由于安全问题或未能预防艾滋病毒而停止了几项此类试验。这些结果强调了对候选nBP的临床前安全性评估的改进的需要，包括对敏感的诊断方法的需要，该方法可以评估nBP诱导的粘膜屏障损伤的程度，并且可以检测可能导致感染易感性增加的炎性细胞和介质的存在。然而，在nBP产品的背景下，用于监测体内粘膜微环境的先进工具的可用性存在关键差距，特别是在易于获得的动物模型中，这些动物模型提供了与人类大体和微观解剖结构和规模相似的粘膜环境。支持模型和技术，提供了一种途径，用于非侵入性和重复体内评估粘膜屏障对nBP预防策略的反应，可以在nBP管道中发挥重要作用，大大有助于开发更安全和有效的产品。在这里，我们假设，在局部应用nBP后，可以通过体内阴道和直肠粘膜的亚细胞和分子成像检测粘膜屏障的微观结构和渗透性的变化。我们的具体目标是开发和验证新的细胞和分子成像技术，用于定量监测和检测敏感的粘膜刺激 并验证这些技术作为粘膜刺激的标志物 以及局部应用nBP后可能与炎症相关的损伤。预期结果包括开发比目前可用于评估nBP作用的更敏感的上皮屏障评估方法，并推进使用绵羊模型对候选nBP进行体内评估，从而有助于开发安全有效的nBP。

DESCRIPTION (provided by applicant): The goal of this project is to develop sensitive and quantitative imaging and cellular approaches for the evaluation of vaginal and rectal mucosal responses to non-vaccine biomedical prevention (nBP) products aimed at preventing acquisition of HIV. A safe and effective nBP strategy could contribute to reducing the incidence of HIV saving millions of lives. While several promising candidate nBPs have emerged in recent years and progressed to clinical trials, a disappointing outcome has been the halting of several such trials due to safety concerns or failure to prevent HIV. These outcomes highlight the need for improved preclinical safety assessment of candidate nBPs, including the need for the sensitive diagnostic methods that can assess the degree of nBP-induced damage in the mucosal barrier and that can detect the presence of inflammatory cells and mediators that may contribute to increased susceptibility to infection. However, there is a critical gap in the availability of advanced tools for monitoring the in vivo mucosal microenvironment in the context of nBP products, particularly in readily accessible animal models that provide a mucosal environment similar to humans in gross and micro-anatomy and scale. Supporting models and technologies that provide an avenue for non-invasive and repeated in vivo assessment of mucosal barrier response to nBP prevention strategies could play an important role in the nBP pipeline, significantly contributing to the development of more safe and effective products. Here, we hypothesize that changes in microstructure and permeability of mucosal barrier can be detected by subcellular and molecular imaging of vaginal and rectal mucosa in vivo, following topical application of nBPs. Our specific aims are to develop and validate novel cellular and molecular imaging techniques for quantitative monitoring and detection of mucosal irritation with sensitivity that is not currently available, and to validate these techniques as a marker of mucosal irritation and injury that can be correlated to inflammation following topical applications of nBP. The expected outcomes include the development of more sensitive epithelial barrier assessment approaches than are currently available for evaluating the effects of nBPs and advancing the use of the sheep model for in vivo assessment of candidate nBPs, thereby contributing to the pipeline for development of safe and effective nBPs.