Peptide Reagents for Detection of Colonic Dysplasia

用于检测结肠发育不良的肽试剂

• 批准号: 7255822
• 负责人: Thomas D Wang
• 金额: --
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-07-01 至 2007-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7255822
• 关键词: Address; Adenomatous Polyps; Affect; Affinity; Amino Acids; Architecture; Area; Award; Bacteriophage M13; Binding; Biopsy; Biopsy Specimen; Bladder; Caliber; Cancer Etiology; Carcinoma; Cervix Uteri; Cessation of life; Clinical; Colon; Colonoscopy; Columnar Cell; Compatible; Data; Depth; Detection; Dimensions; Disease; Dyes; Dysplasia; Early Diagnosis; Endoscopes; Endoscopy; Epithelial; Epithelium; Esophagus; Excision; Fluorescein-5-isothiocyanate; Fluorescence; Future; Genus Cola; Guidelines; Hand; Histopathology; Image; Imaging Techniques; Intestines; Large Intestine Carcinoma; Lesion; Libraries; Light; Lung; Lymph Node Involvement; Malignant - descriptor; Malignant Neoplasms; Methodology; Methods; Microscope; Modeling; Monitor; Morbidity - disease rate; Mucous Membrane; Noise; Numbers; Optical Coherence Tomography; Optical Methods; Optics; Organ; Pan Genus; Patients; Penetration; Peptides; Phage Display; Premalignant; Procedures; Publications; Purpose; Range; Reagent; Recording of previous events; Research; Resected; Resolution; Risk; Sampling Errors; Scanning; Screening procedure; Signal Transduction; Specimen; Stomach; Stratification; Submucosa; Surface; Surgical margins; Techniques; Time; Tissues; Ulcerative Colitis; Work; adenoma; base; colon dysplasia; colorectal cancer screening; cost; fluorescence imaging; gastrointestinal; in vivo; millimeter; mortality; novel; novel strategies; prevent; prototype

DESCRIPTION (provided by applicant): Cancer originating from the epithelium of hollow organs, such as colon, esophagus, stomach, lung, cervix, and bladder, result in over 90% of all deaths by cancer in the U.S. The morbidity and mortality associated with this disease may be prevented by early detection of pre-malignant (dysplastic) mucosa. Confocal microendoscopy is an emerging technique of optical sectioning that can perform real time in vivo histopathology when adequately developed. The dual axes confocal architecture is a novel approach that provides sub-cellular resolution with long working distance, deep tissue penetration, high dynamic range, and scalability to millimeter dimensions with use of post-objective scanning and MEMS micro-mirrors. The long term objectives of this application are to identify peptide reagents that bind preferentially to dysplastic mucosa and to evaluate the clinical utility of confocal microendoscopy to perform in vivo detection, monitoring, and risk stratification of dysplasia present on epithelial surfaces with use of these peptide-dye reagents. This integrated optical methodology will first be demonstrated in colon using the adenoma as a model for dysplasia, and can then be applied to the early detection of other cancers with epithelial origin. The specific aims of this proposal include 1) to identify high affinity peptides that preferentially bind colonic dysplasia with high target-to-background ratio, screen candidate peptides for optimal mucosal binding affinity, and conjugate peptides to FITC; 2) to collect confocal fluorescence images ex vivo from biopsy specimens of colonic mucosa with topically administered peptide-FITC conjugates with the tabletop dual axes prototype, and 3) with the miniature, endoscope compatible dual axes prototype, including characterizing image signal-to-noise ratio, contrast ratio, and resolution. The preferential binding peptides will be identified using a M13 phage display library that expresses linear 7 amino acid peptides with a complexity of 109. Non-specific binding peptides will be removed from the library by panning against non-dysplastic columnar cells in culture and excised specimens of normal colonic mucosa. Dysplasia specific binding peptides will then be identified by panning with freshly resected adenomas. Candidate peptide-FITC reagents will then be administered topically to additional freshly resected adenomas, and then imaged first with the tabletop dual axes confocal microscope, and then with the miniature prototype.

描述（由申请人提供）： 在美国，源自中空器官（如结肠、食道、胃、肺、子宫颈和膀胱）上皮的癌症占所有癌症死亡人数的 90% 以上。与这种疾病相关的发病率和死亡率可以通过早期预防来预防。检测癌前（发育不良）粘膜。共焦显微内窥镜是一种新兴的光学切片技术，如果发展充分，可以进行实时体内组织病理学分析。双轴共焦架构是一种新颖的方法，通过使用后物镜扫描和 MEMS 微镜，可提供亚细胞分辨率、长工作距离、深层组织穿透、高动态范围以及毫米尺寸的可扩展性。本申请的长期目标是鉴定优先与不典型增生粘膜结合的肽试剂，并评估共聚焦显微内窥镜的临床实用性，以使用这些肽对上皮表面上存在的不典型增生进行体内检测、监测和风险分层。染料试剂。这种集成光学方法将首先在结肠中使用腺瘤作为不典型增生模型进行验证，然后可应用于其他上皮来源癌症的早期检测。该提案的具体目标包括1）鉴定以高靶背景比优先结合结肠发育不良的高亲和力肽，筛选最佳粘膜结合亲和力的候选肽，并将肽与FITC缀合； 2) 使用桌面双轴原型，使用局部施用的肽-FITC缀合物，从结肠粘膜活检标本中收集离体共焦荧光图像，以及 3) 使用微型、内窥镜兼容的双轴原型，包括表征图像信噪比比率、对比度和分辨率。将使用 M13 噬菌体展示文库来鉴定优先结合肽，该文库表达复杂性为 109 的线性 7 个氨基酸肽。通过对培养物和切除标本中的非发育不良柱状细胞进行淘选，将非特异性结合肽从文库中去除正常结肠粘膜。然后通过用新鲜切除的腺瘤进行淘选来鉴定不典型增生特异性结合肽。然后将候选肽-FITC 试剂局部施用到另外的新鲜切除的腺瘤上，然后首先使用桌面双轴共聚焦显微镜成像，然后使用微型原型成像。