Fluorescence Lifetime Imaging (FLIm): a method to evaluate colon inflammation in vivo

项目总结/摘要 炎症性肠病（IBD）是胃肠道（GI）疾病的总称，其中慢性炎症性肠病（IBD）是胃肠道（GI）疾病的一种。 炎症及其后遗症显著影响患者的身体健康、生活质量和医疗保健 利用率胃肠道炎症的早期诊断可以促进早期医疗干预，直接影响患者的健康。 患者的预后和生活质量。需要一种能够早期和低水平检测肿瘤的新方法。 胃肠道炎症分级。这个提议的目的是证明荧光寿命成像的可行性 （FLIm）用于检测体内早期结直肠炎症。在不需要外源性标记剂的情况下，FLIm是 对细胞代谢的变化敏感，细胞代谢在炎症过程开始时改变。 我们的具体目标集中在建立FLIM作为一种研究工具，以量化和监测炎症， 组织水平，使用体内鼠结肠作为模型。在目标1中，我们将（子目标1.1）制造侧视图 用于结肠全长的无损、原位和体内腔内成像的内窥镜探头， （子目标1.2）显示FLIm对使用野生型和PPAR-g敲除小鼠的上皮代谢敏感 用抗生素（链霉素）治疗，以产生短暂的生态失调，并用5-阿萨预防抗生素 方面的影响.在目标2中，我们将测试FLIM用于检测早期炎症变化的相关性， 概括了IBD前期的各个方面（高脂肪饮食和抗生素）。使用图像处理和统计分析， 我们将用组织病理学和生物化学测定（H&E，组织缺氧， 细胞内乳酸盐、NAD+/NADH、ADP/ATP、PDH活性）（n = 6只动物/组， 雄性和雌性小鼠）。这项研究产生的广泛的炎症反应将 证明FLIm作为无标记、非破坏性体内管腔内检测的研究工具的灵敏度 并监测宿主对胃肠道炎症的反应。 该研究结果有望为后续R 01型 研究补助金申请，建立在拟议的概念。PI的长期目标是建立FLIM 作为一种非破坏性和无标记的原位和体内介观成像方式，用于研究1）发病机制 2）宿主-微生物群与药理学和饮食的关系 变化，以及3）将该方法转化为用于人体GI的体内内窥镜成像的临床工具 道。我们预计FLIm拟议实施的应用范围包括早期检测 炎症和感染性疾病和癌症的药物和营养的密切监测 胃肠道的治疗

PROJECT SUMMARY/ABSTRACT Inflammatory bowel disease (IBD) is an umbrella term for gastrointestinal (GI) diseases where chronic inflammation and its sequelae significantly impact a patient’s physical health, quality of life, and healthcare utilization. Early diagnosis of GI inflammation could prompt earlier medical intervention with a direct impact on patient’s prognosis and quality of life. There is a need for novel methods capable of detecting early and low- grade GI inflammation. The goal of this proposal is to demonstrate the feasibility of fluorescence lifetime imaging (FLIm) for detecting early colorectal inflammation in vivo. Without requiring exogenous labeling agents, FLIm is sensitive to changes in cellular metabolism, which is altered at the onset of inflammatory processes. Our specific aims focus on establishing FLIm as a research tool to quantify and monitor inflammation at the tissue level using the in vivo murine colon as a model. In Aim 1 we will (sub-aim 1.1) fabricate a side-viewing endoscopic probe for nondestructive, in situ, and in vivo intraluminal imaging of the full length of the colon and (sub-aim 1.2) show that FLIm is sensitive to epithelial metabolism using wild-type and PPAR-g knockout mice treated with antibiotic (streptomycin) to generate transient dysbiosis and with 5-ASA to protect against antibiotic effects. In Aim 2 we will test the relevance of FLIm for detecting early inflammatory changes with a model that recapitulates aspects of pre-IBD (high-fat diet and antibiotics). Using image processing and statistical analysis, we will validate the FLIm parameters with histopathology and biochemical assays (H&E, tissue hypoxia, intracellular lactate, NAD+/NADH, ADP/ATP, PDH activity) performed after necropsy (n = 6 animals/group for both male and female mice). The broad range of inflammatory responses generated with this study will demonstrate the sensitivity of FLIm as a research tool for label-free, nondestructive, in vivo intraluminal detecting and monitoring the host response to GI inflammation. Results from this research are expected to provide convincing preliminary data for subsequent R01-type research grant applications that build on the proposed concept. The long-term goal of the PI is to establish FLIm as a nondestructive and label-free, in situ and in vivo, mesoscopic imaging modality to study 1) pathogenesis and treatment of GI inflammation over time, 2) the host-microbiota relationship with pharmacological and dietary changes, and 3) to translate this approach into a clinical tool for in vivo endoscopic imaging of the human GI tract. We anticipate that the applications of the proposed implementation of FLIm range from early detection of inflammatory and infectious diseases and cancer to the close monitoring of pharmacological and nutritional treatments on the GI tract.