Establishing MAGE-A4/RAD18 as a novel cancer-specific chemotherapeutic target

将 MAGE-A4/RAD18 确立为新型癌症特异性化疗靶点

基本信息

批准号：
10596489
负责人：
Kenneth Hugh Pearce
金额：
$ 39.15万
依托单位：
UNIV OF NORTH CAROLINA CHAPEL HILL
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-04-02 至 2024-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10596489
关键词：
Affect Affinity Antigens Antineoplastic Agents Binding Biological Testing Cancer Model Carcinogenesis Mechanism Cause of Death Cell Survival Cells Cellular Assay Chemicals Chemoresistance Complex DNA Damage DNA Repair DNA Repair Gene DNA Replication Induction Engineering Environmental Risk Factor Genes Genomic Instability Germ Cells Goals Knowledge Libraries Lung Neoplasms Malignant Neoplasms Malignant neoplasm of lung Mission Mitotic Mitotic spindle Molecular Mutagens Normal Cell Nude Mice Oncogenes Oncogenic Pathologic Pathway interactions Peptide Phage Display Library Peptides Permeability Phenotype Physiological Platinum Poison Process Proteins Public Health Research Resistance Role Signal Pathway Signal Transduction Somatic Cell Stress Testing Testis Therapeutic Therapeutic Agents Tissues Topoisomerase II Toxic effect Transgenic Mice Ubiquitin United States National Institutes of Health Work cancer cell cancer therapy carcinogenesis chemotherapeutic agent chemotherapy defined contribution drug discovery druggable target environmental agent experience experimental study improved in vivo inducible Cre inhibitor innovation melanoma mortality neoplastic cell new therapeutic target novel patient prognosis preclinical study preneoplastic cell pressure prevent programs refractory cancer replication stress response screening side effect small molecule targeted treatment taxane therapeutic target therapy resistant tumor tumorigenesis tumorigenic ubiquitin-protein ligase

Affect Affinity Antigens Antineoplastic Agents Binding Biological Testing Cancer Model Carcinogenesis Mechanism Cause of Death Cell Survival Cells Cellular Assay Chemicals Chemoresistance Complex DNA Damage DNA Repair DNA Repair Gene DNA Replication Induction Engineering Environmental Risk Factor Genes Genomic Instability Germ Cells Goals Knowledge Libraries Lung Neoplasms Malignant Neoplasms Malignant neoplasm of lung Mission Mitotic Mitotic spindle Molecular Mutagens Normal Cell Nude Mice Oncogenes Oncogenic Pathologic Pathway interactions Peptide Phage Display Library Peptides Permeability Phenotype Physiological Platinum Poison Process Proteins Public Health Research Resistance Role Signal Pathway Signal Transduction Somatic Cell Stress Testing Testis Therapeutic Therapeutic Agents Tissues Topoisomerase II Toxic effect Transgenic Mice Ubiquitin United States National Institutes of Health Work cancer cell cancer therapy carcinogenesis chemotherapeutic agent chemotherapy defined contribution drug discovery druggable target environmental agent experience experimental study improved in vivo inducible Cre inhibitor innovation melanoma mortality neoplastic cell new therapeutic target novel patient prognosis preclinical study preneoplastic cell pressure prevent programs refractory cancer replication stress response screening side effect small molecule targeted treatment taxane therapeutic target therapy resistant tumor tumorigenesis tumorigenic ubiquitin-protein ligase

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项目摘要

SUMMARY There are fundamental gaps in our understanding of how cancer cells acquire DNA damage-tolerance and evade chemotherapy (termed ‘chemoresistance’). The gaps in our knowledge of DNA damage tolerance (and how it differs between normal and neoplastic cells), limit our ability to kill tumors without causing side effect toxicities to normal healthy tissues. Our long-term goals are to solve the problem of how neoplastic cells tolerate therapy, and identify new molecular vulnerabilities that can be specifically targeted for improved treatment. The objective here is to define a novel tumor-specific mechanism of chemoresistance and to establish its tractability as a therapeutic target. Remarkably, we discovered that Melanoma Antigen-A4 (MAGE-A4, a Cancer/Testes Antigen or 'CTA') is an activating binding partner of the DNA repair protein RAD18 (an E3 ubiquitin ligase). MAGE-A4 is absent from all normal somatic cells but pathologically activates DNA repair in cancer cells. MAGE-A4 also activates the E3 ligase TRIM69 which confers resistance to mitotic spindle poisons. MAGE-A4 expression is associated with poor patient prognosis yet its potential impact on the responsiveness of tumors to chemotherapy in a physiological setting is untested. Based on exciting and compelling preliminary studies, we will test the central hypothesis that MAGE-A4 pathologically reprograms ubiquitin signaling in tumors to confer chemoresistance to genotoxins and spindle poisons. The rationale is that defining the contribution of MAGE-A4 to chemoresistance will allow extraordinarily specific therapeutic strategies that target a unique molecular vulnerability of neoplastic cells. The Specific Aims are: SA1 Define contribution of pathologically-activated DNA repair to chemoresistance in vivo. SA2 Establish chemical tractability of MAGE-A4/RAD18 as a therapeutic target. SA3 Mechanistically define MAGE-A4/TRIM69 functions in mitotic progression and resistance to spindle poisons. In SA1 We will use a new transgenic mouse and orthotopic lung cancer models to determine how MAGE-A4/RAD18 impacts responses to chemotherapy in vivo. In SA2 we will screen peptide phage display libraries to identify sequence motifs that bind MAGE-A4 and disrupt the MAGE-A4/RAD18 interaction. Bioactive MAGE-A4 inhibitor peptides will be tested for anti-neoplastic activity. In SA3 we will define a novel role of MAGE-A4 in regulating TRIM69 and conferring resistance to spindle poisons. We will mechanistically define the MAGE-A4/TRIM69 signaling pathway and establish its role in tolerance of therapeutic taxanes. These experiments will likely establish MAGE-A4 as a druggable target for ameliorating chemoresistance in cancer cells, serving as a crucial gateway in the drug discovery process. The proposed ideas and research are innovative because they are the first studies to test how biological activities of CTAs affect cancer therapy. The proposed work is significant because it will provide new paradigms for chemoresistance due to pathological ubiquitin signaling leading directly to novel targeted therapies for chemoresistant cancer.

概括我们对癌细胞如何获得DNA耐受性和逃避化疗（称为“化学抗性”）。我们了解DNA损伤耐受性的差距（和它在正常细胞和肿瘤细胞之间的不同之处），限制我们杀死肿瘤的能力而不会引起副作用对正常健康组织的毒性。我们的长期目标是解决肿瘤细胞的问题耐受治疗，并确定可以专门针对改进的新分子漏洞治疗。这里的目的是定义一种新型的化学耐药性机制和确定其作为治疗靶标的障碍。值得注意的是，我们发现黑色素瘤抗原A4 （MAGE-A4，癌/睾丸抗原或“ CTA”）是DNA修复蛋白的激活结合伙伴 RAD18（E3泛素连接酶）。所有正常的体细胞都不存在MAGE-A4，但病理激活癌细胞中的DNA修复。 Mage-A4还激活E3连接酶TRIM69，该连接酶Trim69赋予了对有丝分裂的抗性主轴毒物。法师A4表达与患者较差有关，但其潜在影响未经测试的肿瘤对化疗的反应性。基于令人兴奋的引人入胜的初步研究，我们将测试MAGE-A4病理重新编程的中心假设肿瘤中的泛素信号传导与基因毒素和主轴毒物的化学抗性。理由是定义法师A4对化学抗性的贡献将允许特定于特定的治疗针对肿瘤细胞独特的分子脆弱性的策略。具体目的是：SA1定义病理激活的DNA修复对体内化学抗性的贡献。 SA2建立化学物质 MAGE-A4/RAD18作为治疗靶标的障碍性。 SA3机械定义的法师A4/TRIM69 在有丝分裂进展和抗螺旋毒物的抗性中的功能。在SA1中，我们将使用新的转基因小鼠和原位肺癌模型，以确定法师A4/rad18如何影响对体内化学疗法。在SA2中，我们将筛选辣椒噬菌体显示库，以识别序列图案结合MAGE-A4并破坏MAGE-A4/RAD18相互作用。生物活性MAGE-A4抑制剂肽将是测试了抗塑性活性。在SA3中，我们将定义法师A4在确定Trim69和赋予对螺旋毒物的抵抗力。我们将机械定义MAGE-A4/TRIM69信号传导途径并确立其在耐受性紫杉烷耐力中的作用。这些实验可能会建立 MAGE-A4是改善癌细胞化学耐药性的可药物靶标，作为关键网关在药物发现过程中。拟议的思想和研究具有创新性，因为它们是第一个测试CTA的生物学活性如何影响癌症治疗的研究。拟议的工作很重要因为它将由于病理泛素信号传导引起的化学抗性提供新的范式直接用于化学耐药性癌症的新型靶向疗法。