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）：越来越多的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且能够自我更新。在发现 CSC 之前，肿瘤进展的克隆模型获得了大多数支持。该模型假设肿瘤内的每个细胞都具有自我更新的能力，因此，如果在治疗期间没有被杀死，则可以用对治疗具有抗性的亚克隆重新填充肿瘤。 CSC 模型认为肿瘤由细胞层次结构组成。肿瘤内的 CSC 是能够自我再生并共同产生非 CSC 后代的细胞，这些后代构成了剩余肿瘤体积的很大一部分。由于 CSC 是一组肿瘤细胞中的一个亚群，因此它们被附着在细胞外部的两种不同分子标记。我们提出了一种基于抗体的成像系统，该抗体对我们将开发的其中两种标记物具有高亲和力。我们还将开发一种用于小报告分子的第三种抗体，该抗体将用染料标记。该抗体将被分成两半，每一半将与两种细胞表面标记物的抗体融合。当细胞表面标记物与抗体结合时，两半将紧密靠近并形成小报告分子的结合袋。我们将通过在显微镜下观察人类肿瘤细胞上的过程来可视化小报告分子是否结合，从而证明该系统的工作原理。小报告分子将带有染料，使其可视化。下面的目标 1-3 概述了该过程。目标 1. 制备并测试细胞表面标记物和小报告分子的抗体。目标 2 将半抗体与全抗体融合并重新测试以表明结合能力没有受到阻碍。目标 3 将融合的抗体和染料标记的小报告分子添加到人类癌细胞中，并在显微镜下观察过程以确定系统是否可以工作。如果我们能够使这种成像系统发挥作用，就可以在人类癌症动物模型中对其进行测试。如果成功，它也可用于人类，以观察治疗期间和治疗后的 CSC 水平。当开发出抗 CSC 疗法时，这将特别有用。