Phosphorylation-dependent regulation of GLI transcription factors

GLI 转录因子的磷酸化依赖性调节

• 批准号: 10586028
• 负责人: Scott Atwood
• 金额: $ 33.46万
• 依托单位: UNIVERSITY OF CALIFORNIA-IRVINE
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-04-11 至 2025-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10586028
• 关键词: Affect; Allografting; Alternative Therapies; Antibodies; Basal cell carcinoma; Biochemistry; Biological Assay; Brain; Carcinoma; Catalogs; Cell Line; Cellular biology; Cessation of life; Clinical; Cyclic AMP-Dependent Protein Kinases; Data; Databases; Development; Disease; Drug resistance; Erinaceidae; Exhibits; FDA approved; GLI gene; GLI2 gene; Genes; Genetic; Genetic Techniques; Genetic Transcription; Goals; Growth; Health; Hematopoietic Neoplasms; Human; Human Development; In Vitro; Malignant Bone Neoplasm; Malignant Neoplasms; Malignant neoplasm of brain; Malignant neoplasm of prostate; Molecular Biology; Mutation; Oncogenic; Outcome; Pancreas; Pathway interactions; Patients; Persons; Phosphorylation; Phosphotransferases; Prostate; Recurrence; Regulation; Resistance; Signal Pathway; Skin Cancer; Somatic Mutation; Stains; Testing; Transcription Coactivator; Transcriptional Activation; United States; Work; Zinc Fingers; acquired drug resistance; antagonist; atypical protein kinase C; cancer type; cell growth; comparative; drug resistance development; gain of function; hedgehog signal transduction; inhibitor; loss of function; malignant breast neoplasm; malignant stomach neoplasm; mortality; mutant; new therapeutic target; overexpression; protein kinase C iota; receptor; screening; small cell lung carcinoma; smoothened signaling pathway; targeted treatment; therapeutic target; therapy development; transcription factor; tumor; tumor growth

Project Summary Basal cell carcinomas (BCCs) are locally invasive epithelial cancers that affect over 4 million patients a year in the United States and are solely driven by activating mutations in the Hedgehog (HH) pathway. Inappropriate HH pathway activation also drives growth of a variety of cancers including brain, pancreatic, prostate, and small cell lung cancer that account for up to 25% of all human cancer deaths. HH antagonists such as vismodegib are FDA-approved to treat advanced and metastatic BCCs, however, nearly 60% of advanced tumors display inherent vismodegib resistance and 20% of tumors that do respond acquire drug resistance every year. This is a highly relevant issue as advanced BCC cases are estimated to approach 400,000 patients each year, illustrating a critical need to identify therapeutic targets downstream of SMO to suppress HH pathway activity. The consequence of not meeting this need will likely be the inability to treat patients who are resistant to current approved therapies, leading to an increase in mortality for patients inflicted with BCC and other HH-dependent cancers. Our long-term objective is to identify and develop targeted therapeutics to treat drug-resistant HH-driven cancers. The overall objective of this application is to define how GLI is oncogenically activated in HH-driven cancer. Our central hypothesis is that GLI phosphorylation drives transcriptional activation and SMO antagonist-resistant BCC growth, and targeting the signaling pathways that activate GLI will suppress tumor growth. My two specific aims will define 1) how GLI zinc finger phosphorylation and 2) how clinically recurrent GLI mutations promote transcriptional activity, tumor growth, and drug resistance. Defining how GLI is activated in cancer may reveal novel therapeutic targets to treat patients with HH-driven cancers. To achieve these aims, we will use BCC cell lines and allografts that overexpress clinically observed and recurrent GLI mutants to assay for tumor growth in the presence or absence of HH antagonists. We will use the GLI mutants that show increased transcriptional activity to define how GLI is activated in cancer using standard molecular biology, biochemistry, cell biology, and genetic techniques. Our preliminary data has already identified three kinases that regulate GLI activity, and we plan to generate phospho-specific mutants to define when, where, and how each kinase acts on GLI. We will immunofluorescently stain human tumors with appropriate antibodies to verify these pathways operate in humans, and we will perform standard gain- and loss-of-function studies to analyze the pathways involved over the lifetime of the tumor.

项目摘要 基底细胞癌（BCC）是局部侵入性上皮癌，每年影响超过400万患者 美国，仅由刺猬（HH）途径激活突变驱动。不当 HH途径的激活还驱动了包括大脑，胰腺，前列腺和 小细胞肺癌占所有人类癌症死亡的25％。 HH拮抗剂，例如 vismodegib经过FDA批准以治疗高级和转移性BCC，但是近60％的高级 肿瘤表现出固有的vismodegib耐药性和20％的肿瘤，这些肿瘤确实会获得耐药性 每年。这是一个高度相关的问题，因为估计高级BCC病例可接近40万名患者 每年，说明识别SMO下游的治疗靶标的至关重要的需求以抑制HH 途径活动。不满足这一需求的结果可能是无法治疗的 对当前批准的疗法有抵抗力，导致因BCC和BCC造成的患者的死亡率增加 其他HH依赖性癌症。 我们的长期目标是识别和开发有针对性的治疗剂来治疗耐药性HH驱动 癌症。该应用的总体目的是定义GLI在HH驱动的 癌症。我们的中心假设是GLI磷酸化驱动转录激活和SMO 抗拮抗剂的BCC生长，靶向激活GLI的信号通路将抑制肿瘤 生长。我的两个具体目标将定义1）Gli锌指磷酸化以及2）如何临床复发 GLI突变促进转录活性，肿瘤生长和耐药性。定义格利 在癌症中激活的可能揭示了新的治疗靶标，可以治疗HH驱动的癌症患者。 为了实现这些目标，我们将使用过表达临床观察到的BCC细胞系和同种异体移植物，并且 复发性GLI突变体在存在或不存在HH拮抗剂的情况下测定肿瘤生长。我们将使用 显示出增加转录活性的GLI突变体，以定义使用GLI在癌症中使用GLI的使用 标准分子生物学，生物化学，细胞生物学和遗传技术。我们的初步数据 已经确定了调节GLI活性的三种激酶，我们计划生成磷酸特异性突变体 定义何时，何地以及每个激酶在GLI上的作用。我们将与 适当的抗体以验证这些途径在人类中运行，我们将执行标准增益和 功能丧失研究以分析肿瘤寿命中涉及的途径。