High-resolution optical imaging assessment of microbicide toxicity

描述（由申请方提供）：评价杀微生物剂的安全性和有效性需要评估杀微生物剂对宫颈阴道和直肠上皮造成的潜在损伤。可用于局部应用杀微生物剂后组织损伤的内窥镜、快速和定量评估的成像技术和方案的开发可能对杀微生物剂在动物模型和临床研究中的开发和测试产生重大影响。不幸的是，目前的成像技术，例如白色光阴道镜检查或结肠镜检查不能提供上皮损伤的高分辨率图像，并且不能探测上皮损伤和炎症可能明显的表面之下。然而，近年来，已经开发了新的成像技术，其中现在可以在体内以微观分辨率对上皮组织进行高分辨率成像。我们的总体目标是开发一种基于内窥镜成像的方法，该方法可用于1）评估杀微生物剂可能诱导的损伤程度和2）将成像研究结果与HSV-2引起的小鼠宫颈阴道道和直肠生殖器感染的易感性相关联。我们将部署新兴的高分辨率成像模式，包括共聚焦荧光显微镜和光学相干断层扫描（OCT），以表征未治疗的性幼稚小鼠以及用已知刺激性杀微生物剂治疗的小鼠的宫颈，阴道和直肠上皮结构中发生的变化。这些结果将与使用HSV-2感染的良好表征的小鼠模型的感染易感性相关。这将使我们能够基于杀微生物剂诱导的上皮变化评估共聚焦荧光和OCT成像对观察到的HSV-2易感性的预测价值，并注意结果的再现性/一致性。在该项目的第一阶段，我们将展示高分辨率光学成像的能力，以定量评估宫颈阴道和直肠组织对已知杀微生物剂的反应，并测试预测杀微生物剂诱导的宫颈阴道敏感性变化的生物终点的能力。在第二阶段，拟议的基于成像的直肠反应评估将扩展到建立基于图像的标记物和应用杀微生物剂后直肠敏感性之间的相关性。此外，仪器和成像参数将进行优化，使成像协议适用于大型动物模型和人类的宫颈阴道和直肠上皮反应的成像。我们还计划使用开发的成像协议，以评估新的杀微生物剂在小动物模型的新产品开发的性能。

DESCRIPTION (provided by applicant): Evaluation of safety and efficacy of microbicides requires an assessment of potential injury caused by microbicides in the epithelium of cervicovaginal tract and rectum. The development of imaging technology and protocols that can be used for endoscopic, rapid, and quantitative assessment of tissue injury following topical application of microbicides could have significant impact on the development and testing of microbicides in animal models and clinical studies. Unfortunately, current imaging technology such as white light colposcopy or colonoscopy cannot provide high resolution images of epithelial injury and cannot probe below the surface where epithelial injury and inflammation may be evident. However, in recent years, new imaging technology has been developed where it is now possible to perform high resolution imaging of epithelial tissue with microscopic resolution in vivo. Our overall goal is to develop an endoscopic imaged based approach that can be used to 1) assess the degree of injury that may be induced by microbicides and 2) correlate the results of imaging studies to susceptibility to genital infection in mice cervicovaginal tract and rectum caused by HSV-2. We will deploy emerging high resolution imaging modalities including confocal fluorescence microscopy and optical coherence tomography (OCT) to characterize the changes that occur in the architecture of cervical, vaginal, and rectal epithelium of untreated sexually na¿ve mice as well as mice treated with known irritative microbicides. These results will be correlated to susceptibility to infection using a well characterized mouse model of HSV-2 infection. This will allow us to assess the predictive value of confocal fluorescence and OCT imaging for observed HSV-2 susceptibility based on microbicide-induced epithelial changes with attention to reproducibility/consistency of findings. In phase I of this project we will demonstrate the capabilities of high resolution optical imaging to quantitatively assess the response of cervicovaginal and rectal tissue to known microbicides and test the ability to predict the biological end point of microbicide-induced changes in susceptibility in cervicovaginal tract. In phase II, the proposed imaged based assessment of rectal response will be extended to establish correlation between image-based markers and rectal susceptibility following application of microbicides. Furthermore, instrumentation and imaging parameters will be optimized to make the imaging protocol suitable for imaging of cervicovaginal and rectal epithelial response in large animal models and humans. We also plan to use the developed imaging protocol to assess the performance of novel microbicides in small animal models as new products are developed.