Human Islet Antigen Discovery and Imaging

人类胰岛抗原的发现和成像

• 批准号: 6576437
• 负责人: Paul E Harris
• 金额: $ 16.35万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-09-30 至 2004-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6576437
• 关键词: NOD mouse; SCID mouse; antibody specificity; antigen antibody reaction; bioimaging /biomedical imaging; electron microscopy; gene expression; genetic library; genetic markers; human tissue; immunoglobulin G; immunoprecipitation; insulin dependent diabetes mellitus; magnetic resonance imaging; method development; microarray technology; pancreas imaging /visualization; pancreatic islet transplantation; polymerase chain reaction; radiotracer; reagent /indicator; serology /serodiagnosis; surface antigens; tissue /cell culture; transplantation immunology; western blottings

DESCRIPTION (provided by applicant): The overall goal of our studies is to develop reagents suitable for imaging human islets in the context of islet transplantation or type 1 diabetes. In this islet marker discovery program, we will borrow heavily from tumor immunology using the antigen mining techniques of gene expression analysis, bioinformatics, serology and protein chemistry. In Specific Aim One, we combine these methods to characterize the cell surface of human islet tissue. We will use two convergent strategies to identify candidate islet specific markers. In part A, transcript profiling will be used to obtain a map of human islet, exocrine pancreas and liver tissue gene expression. This information will allow us to generate Venn diagrams of candidate markers that are expressed by human islets and not by the exocrine component of the pancreas and not by the liver. Alternate nucleic acid based methods will be used to validate the tissue specificity and distribution of candidate markers determined by transcript profiling. In part B, we will use an adaptation of the serological analysis of recombinant cDNA expression libraries (SEREX) technique (a.k.a., antibody screening of phage expression libraries) to identify islet cell surface associated molecules that may be useful in imaging. Expression libraries will be created in a unique manner as to remove genes that are expressed both in islets and in the exocrine component of the pancreas (or liver). Candidate marker proteins will then be expressed in bacterial systems for the generation of specific antisera. In Specific Aim Two, we will characterize the biodistribution and uptake kinetics of radiolabeled antibodies recognizing candidate markers. We will explore and compare the ability of Dithizone based and immunoglobulin (Ig) based radiolabel imaging reagents to detect islet tissue in vivo. In the last phase of the proposal we shall determine whether these reagents can be used as magnetic resonance imaging (MRI) contrast reagents. In these preclinical studies we will determine whether human islets, transplanted into an immunodeficient animal model, can be detected using reagents modified for use in non-invasive magnetic resonance (MR) imaging studies. We have drawn together an interdisciplinary research team of cell biologists, tumor antigen biologists, transplant surgeons and medical physicists with a common goal of developing of imaging reagents and techniques that distinguish islets from their surrounding tissue.

描述（由申请人提供）：我们的研究的总体目标是开发适合在胰岛移植或1型糖尿病的背景下成像人类胰岛的试剂。在这个小岛标记发现程序中，我们将使用基因表达分析，生物信息学，血清学和蛋白质化学的抗原挖掘技术从肿瘤免疫学中借用。在特定目标中，我们结合了这些方法以表征人类胰岛组织的细胞表面。我们将使用两种收敛策略来识别候选胰岛特定标记。在A部分中，转录物分析将用于获得人类胰岛，外分泌胰腺和肝组织基因表达的图。这些信息将使我们能够生成由人类胰岛表达的候选标记的维恩图，而不是由胰腺的外分泌成分而不是肝脏。基于核酸的替代方法将用于验证通过转录分析确定的候选标记的组织特异性和分布。在B部分中，我们将使用重组cDNA表达文库（SEREX）技术的血清学分析（又称噬菌体表达文库的抗体筛选）的适应性来识别可能在成像中有用的胰岛细胞表面相关分子。表达式库将以独特的方式创建，以消除胰腺（或肝脏）的外分泌成分中既表达的基因。然后，候选标记蛋白将在细菌系统中表达，以生成特定的抗血清。在特定目标二中，我们将表征识别候选标记的放射性标记抗体的生物分布和摄取动力学。我们将探索和比较基于二硫酮的基于二硫酮和免疫球蛋白（IG）的放射性标映成像试剂在体内检测胰岛组织的能力。在提案的最后阶段，我们应确定这些试剂是否可以用作磁共振成像（MRI）对比试剂。在这些临床前研究中，我们将使用修饰的试剂用于非侵入性磁共振（MR）成像研究来检测到移植到免疫缺陷动物模型的人类胰岛。我们将细胞生物学家，肿瘤抗原生物学家，移植外科医生和医学物理学家的跨学科研究团队汇总在一起，其共同目标是开发成像试剂和技术，这些试剂和技术将胰岛与周围的组织区分开来。