ANTI-PDZ DOMAIN ANTIBODIES ENHANCE THE EFFICACY OF RADIOTHERAPY

抗 PDZ 结构域抗体增强放射治疗的疗效

基本信息

批准号：
9198674
负责人：
Sriram Devanathan
金额：
$ 21.69万
依托单位：
MEDICAL GUIDANCE SYSTEMS, LLC
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-09-19 至 2019-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9198674
关键词：
Affinity Antibodies Antineoplastic Agents Apoptosis Binding Binding Proteins Biological Assay Blocking Antibodies Breast Cancer Prognosis Cell Line Cell Lineage Cell Proliferation Cell Survival Cell membrane Cell surface Cells Clinical Trials Cytostatics Cytotoxic Chemotherapy Data Development Docking Effector Cell Enhancing Antibodies Enzymes Epitopes Evaluation Exhibits Exposure to Fc Receptor Fibroblasts Flow Cytometry Future G-Protein-Coupled Receptors GRP78 gene Gene Silencing Glioblastoma Goals Human Hybridomas IgG1 Immune Immunoglobulin Fragments Immunoglobulin G In Vitro Interruption Ionizing radiation Lead Lung Malignant Neoplasms Malignant neoplasm of lung Measures Mediating Molecular Molecular Chaperones Monoclonal Antibodies Motor Mus Normal tissue morphology Phage Display Phase Play Population Production Proteins Radiation Radiation therapy Research Role Scaffolding Protein Signal Pathway Signal Transduction Signal Transduction Pathway Signaling Protein Small Business Innovation Research Grant Specificity Surface Taxes Technology Therapeutic Effect Toxic effect Tumor Volume Xenograft Model base biological adaptation to stress cancer cell cancer therapy comparative efficacy cytotoxic cytotoxicity drug development humanized antibody improved in vivo mouse model neoplastic cell novel strategies optical imaging overexpression polyclonal antibody response rho safety testing tax Gene Products tumor tumor growth tumor xenograft

项目摘要

Poor prognosis cancers including cancers of the lung and glioblastoma show robust overexpresion of Tax Interacting Protein-1 (TIP1). The proposed research is a novel approach to cancer drug development in which we target this chaperone protein that binds to signal transduction proteins that enhance cell viability following cytotoxic therapy. Stress responses in cancer cells are exaggerated over that of normal tissues. Examples are signal transduction pathways that include GRP78, PKC, PLC, Rho and others. Mechanisms by which these proteins dock on the plasma membrane in response to ionizing radiation include motor and scaffold proteins. One scaffold protein that caps the carboxyl terminus of plasma membrane-associated signaling proteins, TIP1 is overexpressed in cancer and translocates to the surface of the cell membrane of cancer cells following exposure to ionizing radiation. The functional domain of TIP1 is the PDZ binding domain which binds enzymes that regulate cell viability such as PLC, PKC, GPCR and Rho. Our preliminary data show that gene silencing of TIP1 and blocking antibodies that bind to the PDZ domain of TIP1 enhance radiation-induced cytotoxicity in cancer but not normal tissues. Anti-PDZ-domain antibodies injected into mice bearing irradiated lung cancer bind specifically to cancer and substantially enhance tumor growth delay over that of controls. We will study antibodies that enhance cytotoxity and improve tumor control. One mechanism by which the anti-PDZ domain antibodies enhance the efficacy of radiotherapy is through interruption of cell viability signal transduction pathways and subsequent programmed cell death specifically in cancer. We will compare antibody fragment (scFv) to whole IgG antibodies that bind to the PDZ domain of TIP1 and determine which antibodies achieve cancer specific binding and enhancement of the efficacy of radiotherapy in cancer without normal tissue toxicity. We will determine the lead human anti-TIP1 antibody fragment that achieves cancer specific binding and cytotoxicity in cancer cells. Our preliminary data show that assays which measure the interaction between anti-TIP1 antibodies and radiation include the clonogenic, and apoptosis assays. We will compare our lead antibodies that are specific to the PDZ domain on TIP1 to determine which is the most cancer specific and cytotoxic. We will determine the efficacy of anti-TIP1/PDZ IgG compared to anti-TIP1/PDZ scFv antibodies in mouse models of cancer. We will determine the role of immune effector cells in the cancer response to anti-TIP1 antibodies. We Hypothesize that IgG could further enhance the efficacy of radiotherapy by activating immune effector cells. We will study mouse models that lack Fc receptors and do not activate antibody mediated cancer cytotoxicity. These mouse models will allow us to determine whether the efficacy of the anti-TIP1 antibodies is a direct effect on cancer cells or whether immune effector cell activation contributes to the therapeutic effect.

预后不良的癌症（包括肺癌和胶质母细胞瘤）显示出 Tax 的强烈过度表达相互作用蛋白-1 (TIP1)。拟议的研究是一种癌症药物开发的新方法，其中我们的目标是这种伴侣蛋白，它与信号转导蛋白结合，从而增强细胞活力细胞毒疗法。癌细胞的应激反应比正常组织的应激反应更强烈。例子是信号转导通路包括 GRP78、PKC、PLC、Rho 等。这些机制响应电离辐射而停靠在质膜上的蛋白质包括运动蛋白和支架蛋白。一种覆盖质膜相关信号蛋白羧基末端的支架蛋白 TIP1 在癌症中过度表达并易位至癌细胞的细胞膜表面暴露于电离辐射。 TIP1的功能域是结合酶的PDZ结合域调节细胞活力，如 PLC、PKC、GPCR 和 Rho。我们的初步数据表明，基因沉默 TIP1 和与 TIP1 PDZ 结构域结合的阻断抗体可增强辐射诱导的细胞毒性癌症但不是正常组织。将抗 PDZ 结构域抗体注射到患有辐射肺癌的小鼠体内与癌症特异性结合，与对照相比显着增强肿瘤生长延迟。我们将学习增强细胞毒性并改善肿瘤控制的抗体。抗 PDZ 结构域的一种机制抗体增强放疗疗效是通过中断细胞活力信号转导途径和随后的程序性细胞死亡，特别是在癌症中。我们将比较抗体片段 (scFv) 到与 TIP1 的 PDZ 结构域结合的完整 IgG 抗体，并确定哪些抗体实现癌症特异性结合并增强无正常组织癌症的放射治疗效果毒性。我们将确定可实现癌症特异性结合的先导人类抗 TIP1 抗体片段，并癌细胞的细胞毒性。我们的初步数据表明，测量抗 TIP1 之间相互作用的测定抗体和辐射包括克隆形成和细胞凋亡测定。我们将比较我们的先导抗体特异性针对 TIP1 上的 PDZ 结构域，以确定哪个是最具癌症特异性和细胞毒性的。我们将确定在小鼠癌症模型中，抗 TIP1/PDZ IgG 与抗 TIP1/PDZ scFv 抗体的功效比较。我们将确定免疫效应细胞在抗 TIP1 抗体的癌症反应中的作用。我们假设 IgG 可以通过激活免疫效应细胞进一步增强放疗疗效。我们将研究小鼠模型缺乏 Fc 受体并且不会激活抗体介导的癌症细胞毒性。这些鼠标模型将使我们能够确定抗 TIP1 抗体的功效是否对癌细胞有直接作用，或者是否具有免疫作用效应细胞的激活有助于治疗效果。