SELECTIVELY IMAGING CANCER STEM CELLS

选择性成像癌症干细胞

基本信息

批准号：
8095997
负责人：
Wilson Barry Edwards
金额：
$ 16.48万
依托单位：
UNIVERSITY OF PITTSBURGH AT PITTSBURGH
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-09-26 至 2013-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8095997
关键词：
Affinity Animal Model Antibodies Antibody Affinity Antigens Automobile Driving Bacteriophages Binding Binding Sites Bispecific Antibodies CD44 Antigens CD44 gene Cell model Cell surface Cells Chimeric Proteins Contrast Media Dyes Flow Cytometry Fluorescence Microscopy Fluorescent Dyes Haptens Health Human Image Imagery Imaging Techniques Immunoglobulin Fragments Knowledge Label Libraries Ligation Lobe Malignant Neoplasms Microscope Modeling Molecular Biology Techniques Molecular Weight Natural regeneration Phage Display Plant Roots Population Process Property Proteins Radio Radioisotopes Reporting Resistance Role Structure System Testing Tracer Ursidae Family Work antigen binding base cancer cell cancer imaging cancer stem cell cancer therapy driving force fluorophore human subject in vivo killings neoplastic cell novel self-renewal stem cell therapy therapy resistant tumor tumor growth tumor progression tumorigenesis

项目摘要

DESCRIPTION (provided by applicant): Evidence is mounting that tumor growth is driven by cancer stem cells (CSCs).These cells are believed to act as the driving force for tumor growth because they are resistant to therapy and are able to self renew. Prior to the discovery of CSCs, the clonal model of tumor progression held the majority of support. This model posits that every cell within a tumor has the capacity for self-renewal and therefore, if not killed during therapy, can repopulate the tumor with subclones resistance to the therapy. The CSC model maintains that a tumor consists of a hierarchy of cells. The CSCs within the tumor are the cells that are able to regenerate themselves and co-produce non-CSC progeny which make up a good deal of the remaining tumor bulk. Because CSCs are a sub population within a group of tumor cells, they are marked with two different molecules that are attached to the outside of the cell. We propose an imaging system based on antibodies that have high affinity for two of these markers which we will develop. We will also develop a third antibody for a small reporting molecule that will be labeled with a dye. This antibody will be divided in half and each half will be fused to the antibodies for the two cell surface markers. When the cell surface markers are bound by the antibodies the two halves will be brought in close proximity and form a binding pocket for the small reporting molecule. We will prove this system works by visualizing whether the small reporting molecule binds by viewing the process on human tumor cells in a microscope. The small reporting molecule will bear a dye that will enable its visualization. The process is outlined below in Aims 1-3. Aim 1. Prepare and test antibodies to the cell surface markers and the small reporting molecule. Aim 2 Fuse the half-antibodies to the whole antibodies and retest to show that binding ability has not been hampered. Aim 3 Add the fused antibodies and dye-labeled small reporting molecule to the human cancer cells and view the process under the microscope to determine whether the system can work. If we are able to make this imaging system work, it can then be tested in animal models of human cancer. If successful, it also be used in humans to view the levels of CSCs during and after therapy. This will be particularly useful when anti-CSC therapies are developed.

描述（由申请人提供）：有证据表明肿瘤生长是由癌症干细胞（CSC）驱动的。这些细胞被认为是肿瘤生长的驱动力，因为它们对治疗有抵抗力并能够自我更新。在发现CSC之前，肿瘤进展的克隆模型拥有大多数支持。该模型认为，肿瘤中的每个细胞都具有自我更新的能力，因此，如果在治疗过程中未杀死，则可以用亚克隆对治疗的耐药性重新填充肿瘤。 CSC模型认为，肿瘤由细胞的层次结构组成。肿瘤内的CSC是能够再生并共同生产非CSC后代的细胞，这些细胞构成了其余肿瘤的大量。由于CSC是一组肿瘤细胞中的亚种群，因此它们具有连接到细胞外部的两个不同分子。我们提出了一个基于对这些标记中的两个标记具有高亲和力的抗体的成像系统。我们还将开发用于将染料标记的小报告分子的第三抗体。该抗体将分为一半，每半将融合到两个细胞表面标记的抗体。当细胞表面标记被抗体结合时，将两半将近距离接近，并形成一个小报告分子的结合袋。我们将通过可视化小报告分子是否通过观察显微镜中人类肿瘤细胞的过程来可视化来证明该系统有效。小型报告分子将带有一种染料，可以使其可视化。该过程在AIMS 1-3中概述。 AIM 1。准备并测试对细胞表面标记和小报告分子的抗体。 AIM 2将半抗体融合到整个抗体中，并重新测试以表明结合能力尚未受到阻碍。 AIM 3向人类癌细胞添加融合抗体和染料标记的小报告分子，并查看显微镜下的过程以确定系统是否可以工作。如果我们能够使该成像系统起作用，则可以在人类癌症的动物模型中进行测试。如果成功的话，它也可以在人类中使用，以查看治疗期间和之后的CSC水平。当开发抗CSC疗法时，这将特别有用。