Peptide Reagents for Detection of Colonic Dysplasia

用于检测结肠发育不良的肽试剂

• 批准号: 7589439
• 负责人: Thomas D Wang
• 金额: $ 6.98万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 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 2009-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7589439
• 关键词: Adenomatous Polyps; Affinity; Amino Acids; Architecture; Award; Bacteriophage M13; Binding; Biopsy Specimen; Bladder; Caliber; Cervix Uteri; Cessation of life; Clinical; Colon; Columnar Cell; Compatible; Data; Depth; Detection; Dimensions; Disease; Dyes; Dysplasia; Early Diagnosis; Endoscopes; Epithelial; Epithelium; Esophagus; Fluorescein-5-isothiocyanate; Fluorescence; Future; Genus Cola; Histopathology; Image; Libraries; Lung; Malignant Neoplasms; Methodology; Microscope; Modeling; Monitor; Morbidity - disease rate; Mucous Membrane; Noise; Optical Methods; Optics; Organ; Pan Genus; Penetration; Peptides; Phage Display; Premalignant; Publications; Purpose; Range; Reagent; Research; Resected; Resolution; Risk; Scanning; Signal Transduction; Specimen; Stomach; Stratification; Surface; Techniques; Time; Tissues; Work; adenoma; base; colon dysplasia; colorectal cancer screening; fluorescence imaging; 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）： 源自空心器官上皮的癌症，例如结肠，食道，胃，肺，子宫颈和膀胱，导致美国癌症的所有死亡人数超过90％，可以通过早期发现与这种疾病相关的发病率和死亡率来预防，并通过早期检测到预防性（pplasticatistrastic）mucosa。共聚焦微观镜检查是一种光学切片的新兴技术，可以在充分发育后实时在体内组织病理学。双轴共聚焦结构是一种新颖的方法，可提供较长的工作距离，深层组织穿透，高动态范围和毫米尺寸的可扩展性，并通过使用后尺寸扫描和MEMS微型练习。本应用的长期目标是鉴定优先结合与发育不良粘膜的肽试剂，并评估共焦微观镜检查的临床实用性，以在体内检测，监测和使用这些肽-Dye Reegents上皮表面上存在性异常的风险分层进行体内检测，监测和风险分层。这种综合的光学方法论将首先在结肠中使用腺瘤作为发育不良的模型证明，然后可以应用于其他具有上皮起源的癌症的早期检测。该提案的具体目的包括1）确定优先结合具有高目标与背景比的结肠发育不良的高亲和力肽，用于最佳粘膜结合亲和力的筛查候选肽，以及偶联肽与FITC； 2）从局部施用的肽-FITC共轭物与桌面双轴原型和3）中，并具有微型，内窥镜兼容轴兼容轴兼容双重轴原型，包括局部给药的肽 - 纤维 - 偶联物，包括局部给予的肽-FITC偶联物，从而从结肠粘膜的活检标本中收集共聚焦荧光图像。将使用M13噬菌体展示库来鉴定优先结合肽，该肽表达线性7氨基酸肽的复杂性为109。非特异性结合肽将通过培养中的非塑性柱细胞在培养物中的非塑性柱细胞（并切除正常结肠粘液菌）来清除文库。然后，将通过新鲜切除的腺瘤平键来鉴定出异常特异性结合肽。然后将候选肽-FITC试剂局部施用到其他新鲜切除的腺瘤，然后首先用桌面双轴共聚焦显微镜成像，然后再用微型原型进行成像。