Probe-based two photon microscopy for functional, label-free early cancer diagnosis

基于探针的双光子显微镜用于功能性、无标记早期癌症诊断

• 批准号: 10030979
• 负责人: ADELA BEN-YAKAR
• 金额: $ 74.7万
• 依托单位: TUFTS UNIVERSITY MEDFORD
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-07-01 至 2024-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10030979
• 关键词: Academia; Algorithms; Area; Benign; Biological Markers; Biopsy; Bladder; Breast; Caliber; Cancer Diagnostics; Cancer Patient; Cancerous; Carcinoma; Cervical; Cervix Uteri; Cessation of life; Clinic; Clinical; Clinical Research; Clinical Treatment; Collection; Colon; Colposcopy; Consumption; Custom; Detection; Development; Diagnosis; Diagnostic; Diagnostic Procedure; Early Diagnosis; Ensure; Epithelial; Epithelium; Esophagus; Evaluation; Fiber; Goals; Gold; Hand; Histologic; Histopathology; Human; Image; Image Analysis; In Situ; Inflammation; Label; Lasers; Lesion; Location; Lung; Malignant Neoplasms; Measurement; Medical center; Metabolic; Metabolism; Metaplasia; Methods; Mitochondria; Morphology; NADH; Nature; Nuclear; Optics; Organ; Outcome; Oxidation-Reduction; Pain; Patients; Physicians; Process; Property; Protocols documentation; Reporting; Resolution; Sensitivity and Specificity; Signal Transduction; Site; Skin; Specificity; Survival Rate; System; Techniques; Testing; Thick; Time; Tissue imaging; Tissues; Training; Triage; Variant; Visualization; austin; base; cancer care; cancer diagnosis; care costs; cervical biopsy; clinical Diagnosis; cost; design; diagnostic accuracy; effective therapy; experience; first-in-human; fluorescence imaging; imaging platform; imaging probe; imaging system; improved; in vivo; in vivo imaging; in vivo imaging system; innovation; miniaturize; multiphoton imaging; outcome forecast; portability; precision medicine; premalignant; psychologic; quantitative imaging; submicron; technology development; tissue biomarkers; two photon microscopy; two-photon

Improvements in the detection of cancerous changes at the pre-invasive stage have the potential to make a significant impact in the prognosis and treatment of most cancer patients. Despite important advances in our understanding of cancer pathobiology, the most prevalent diagnostic methods continue to rely on low magnification tissue visualization, followed by biopsy and histopathology. This process is invasive, often limited in its sensitivity and/or specificity, time-consuming, and relies heavily on the expertise of highly trained physicians, who in turn exploit primarily morphological tissue changes for their assessments. These limitations present barriers to effective treatment and raise monetary and psychological costs. Our long-term objective is to transform pre- and early epithelial cancer diagnosis through the use of functional (metabolic) and morphological tissue biomarkers that are extracted non-invasively, automatically, and in near real time from fiber-probe-based endogenous two-photon (2P) images. In this proposal, we aim to establish and validate such measurements and biomarkers for the detection of human cervical pre-cancers in vivo. The cervix is an ideal organ for developing and testing our approach as it relaxes some of the size limitations presented for endoscopic applications, enabling us to focus on demonstrating the principles of this innovative platform. In addition, we expect that our proposed methods will enable significant near-term improvements in the sensitivity and specificity of detection of cervical pre-cancerous lesions during colposcopy and triage with therapy. To achieve our goals, we have forged a strong partnership among colleagues in academia and the clinic, leveraging strengths and expertise of multiple teams. Specifically, we will exploit our experience in laser development (Xu, Cornell) and multiphoton imaging probe design (Ben-Yakar, UT Austin) to develop a probe-based 2P imaging system that is portable and enables fast image acquisition throughout the cervical epithelium depth with submicron resolution (Aim 1). The final design specifications will be optimized to enable automated, near-real time analysis of images that provides quantitative metrics of metabolic function and morphology (Aim 2). In particular, we will assess: a) multiple quantitative biomarkers of cellular metabolism based on redox ratio and mitochondrial organization parameters extracted from endogenous NAD(P)H and FAD 2P excited fluorescence images of the epithelium, and b) morphological metrics associated with depth-dependent variations of the nuclear to cytoplasmic area ratio and epithelial thickness (Georgakoudi, Tufts). The real time algorithms will be established using freshly excised normal and pre-cancerous human cervical tissues (Aim 2). They will be tested and further optimized in vivo when the innovative 2P imaging platform will be used to perform the first-in-human 2P probe-based imaging during colposcopy (Thieu/Genega, Tufts Medical Center) (Aim3). We expect these studies will provide high sensitivity and specificity of cervical pre-cancer detection and will enable further studies that have the potential to change the paradigm of diagnosis and ultimately prognosis for a broad range of cancers that are accessible via a probe.

在侵入阶段发现癌性变化的检测的改善有可能使 对大多数癌症患者的预后和治疗的重大影响。尽管我们的重要进展 了解癌症病理生物学，最普遍的诊断方法继续依赖 放大组织可视化，然后进行活检和组织病理学。这个过程是侵入性的，通常有限 在其敏感性和/或特异性，耗时，并在很大程度上取决于训练有素的专业知识 医师反过来剥削主要是形态组织的评估变化。这些限制 目前的有效治疗障碍并提高货币和心理成本。我们的长期目标是 通过使用功能（代谢）和形态学转化前和早期上皮癌的诊断 从基于纤维探针的，无创，自动且几乎实时地提取的组织生物标志物 内源性两光子（2p）图像。在此提案中，我们旨在建立和验证此类测量和 在体内检测人类宫颈前癌的生物标志物。子宫颈是发展的理想器官 并测试我们的方法，因为它放松了内窥镜应用的一些尺寸限制， 使我们能够专注于展示这个创新平台的原理。此外，我们希望我们的 提出的方法将在检测的敏感性和特异性方面实现显着的近期改善 在阴道镜检查和治疗分类过程中的颈前病变。为了实现我们的目标，我们有 在学术界和诊所之间建立了牢固的合作伙伴关系，利用优势和专业知识 多个团队。具体而言，我们将利用激光开发（XU，Cornell）和Multiphoton的经验 成像探针设计（Ben-Yakar，UT Austin）开发一种基于探针的2P成像系统，该系统是便携式的 通过亚微米分辨率在整个宫颈上皮深度中启用快速图像采集（AIM 1）。这 最终设计规范将被优化，以实现提供的图像的自动化，近乎真实的时间分析 代谢功能和形态学的定量指标（AIM 2）。特别是，我们将评估：a）多个 基于氧化还原比和线粒体组织参数的细胞代谢的定量生物标志物 从上皮的内源性NAD（P）H和FAD 2P激发荧光图像中提取，B） 核与细胞质面积比的深度依赖性变化相关的形态指标和 上皮厚度（Georgakoudi，簇）。实时算法将使用新鲜切除 正常和癌前的人宫颈组织（AIM 2）。它们将在体内进行测试并进一步优化 创新的2P成像平台将用于执行第一个人类2p探针基于探针的成像 阴道镜检查（Thieu/Genega，Tufts医疗中心）（AIM3）。我们预计这些研究将提供高灵敏度 和宫颈前检测的特异性，并将实现进一步的研究 可以通过探针进入的广泛癌症的诊断范式和最终预后。