PERSONALIZED THERAPY FOR p16-DEFICIENT MELANOMA

p16 缺乏的黑色素瘤的个性化治疗

基本信息

批准号：
9933633
负责人：
Romi Gupta
金额：
$ 7.43万
依托单位：
UNIVERSITY OF ALABAMA AT BIRMINGHAM
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-08-01 至 2020-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9933633
关键词：
Affinity Chromatography Area Biochemical Genetics CDKN2A gene Cell Culture Techniques Cell Cycle Cells Cessation of life Clinical Combined Modality Therapy Complement Data Disease Doxycycline Epigenetic Process Gene Expression Profiling Genes Genetic Genetic Predisposition to Disease Genetic Transcription Genetic study Genome Genomic approach Growth Human Immunotherapy Maintenance Mass Spectrum Analysis Mediating Medical Melanoma Cell Metastatic Melanoma Molecular Mutation Neoplasm Metastasis Organ Pathogenesis Pathway interactions Patients Pharmacology Phosphotransferases Proteins Proteomics Public Health RNA Interference RNA interference screen Regulation Role Skin Cancer Testing The Cancer Genome Atlas Therapeutic Tumor Suppressor Proteins Tumor-Suppressor Gene Inactivation United States base cancer cell cancer type effective therapy experimental study functional genomics improved in vivo in vivo Model inhibitor/antagonist innovation knock-down melanoma mouse model novel personalized medicine response small hairpin RNA small molecule inhibitor transcription factor transcriptome tumor tumor growth

项目摘要

Project Summary Melanoma account for over 10,000 deaths annually in the United States alone and current therapies for advanced-stage melanoma do not provide durable clinical benefit. Therefore, improved molecular understanding of melanoma is essential for developing effective therapies. According to The Cancer Genome Atlas (TCGA) melanoma data over 45% melanoma lack tumor suppressor p16. However, there is no specific personalized treatment option available for p16-deficient melanoma. Therefore, we performed an innovative large-scale druggable genome RNAi screen and identified Cell division cycle 7 (CDC7) kinase as a candidate that is necessary for the survival of p16-deficient melanoma. shRNA-induced knockdown of CDC7 inhibited p16-deficient melanoma tumors and metastases in vivo. Our central hypothesis is that p16-deficient melanoma depends upon CDC7 for their survival and CDC7 inhibition will be of therapeutic value for treating p16-deficient melanoma. The overall objective is to determine the effect of genetic and pharmacological inhibition of CDC7 on melanoma tumor growth and metastasis and understand why CDC7 is necessary for p16-deficient melanoma. Specifically, in Aim 1, we will determine the effect of genetic and pharmacological inhibition of CDC7 on melanoma tumor growth and metastasis. To this end, will determine the role of CDC7 in tumor growth and metastasis in the context of p16-deficiency. Additionally, using XL413, a small molecule inhibitor of CDC7, alone or in combination with BRAFV600E inhibitor vemurafenib (for melanoma co-harboring both BRAFV600E mutation and p16-deficiency), we will determine if pharmacological inhibition of CDC7 alone or in combination with BRAFV600E inhibitors can be employed to effectively treat p16-deficient melanoma in vivo. The experimental approach will utilize mouse models of melanoma tumor growth and metastasis and will utilize established and melanoma patient-derived short-term melanoma cultures available from Yale SPORE in Skin Cancer. In Aim 2, we will determine why CDC7 is necessary for the survival of p16-deficient melanoma. Using transcriptome-wide gene expression profiling and Tandem Affinity Purification and mass spectrometry- based proteomics approaches, we find that the p16-deficient melanoma cells show activation of Unfolded Protein Response (UPR) pathway and that CDC7 maintains the activity of UPR pathway potentially via transcription factor HOXA1 to promote melanoma tumor growth and metastasis. Therefore, we will determine how CDC7 regulates UPR pathway via HOXA1 in p16-deficient melanoma cells. Additionally, we will test whether CDC7-loss induced tumor growth and metastasis inhibition of p16-deficient melanoma cells can be rescued by restoring HOXA1 expression and the UPR pathway activity. Collectively, we expect to uncover a novel CDC7-driven druggable genetic vulnerability pathway that can be targeted for effective personalized therapy of p16-deficient melanoma, including for highly aggressive metastatic melanoma.

项目概要仅在美国，每年就有超过 10,000 人因黑色素瘤死亡，目前的治疗方法晚期黑色素瘤不能提供持久的临床益处。因此，改进的分子了解黑色素瘤对于开发有效的治疗方法至关重要。根据癌症基因组 Atlas (TCGA) 黑色素瘤数据显示超过 45% 的黑色素瘤缺乏肿瘤抑制因子 p16。但并没有具体规定针对 p16 缺乏的黑色素瘤提供个性化治疗选择。因此，我们进行了一次创新性的大规模可药物基因组 RNAi 筛选并确定细胞分裂周期 7 (CDC7) 激酶作为候选者这对于缺乏 p16 的黑色素瘤的生存是必需的。 shRNA 诱导的 CDC7 敲低受到抑制 p16 缺陷的黑色素瘤肿瘤和体内转移。我们的中心假设是 p16 缺陷黑色素瘤的生存依赖于 CDC7，而 CDC7 抑制对于治疗黑色素瘤具有治疗价值 p16 缺乏的黑色素瘤。总体目标是确定遗传和药理学的影响 CDC7 对黑色素瘤生长和转移的抑制作用，并了解为什么 CDC7 对于黑色素瘤肿瘤生长和转移是必需的 p16 缺乏的黑色素瘤。具体来说，在目标 1 中，我们将确定遗传和药理学的影响 CDC7对黑色素瘤生长和转移的抑制作用。为此，将确定CDC7在 p16 缺乏情况下的肿瘤生长和转移。此外，使用小分子 XL413 CDC7 抑制剂，单独或与 BRAFV600E 抑制剂维莫非尼 (vemurafenib) 组合（用于黑色素瘤共同携带） BRAFV600E 突变和 p16 缺陷），我们将确定是否单独对 CDC7 进行药理学抑制或与 BRAFV600E 抑制剂联合使用可有效治疗 p16 缺陷型黑色素瘤体内。该实验方法将利用黑色素瘤肿瘤生长和转移的小鼠模型，并将利用耶鲁 SPORE 提供的已建立的黑色素瘤患者来源的短期黑色素瘤培养物皮肤癌。在目标 2 中，我们将确定为什么 CDC7 对于 p16 缺陷型黑色素瘤的生存是必需的。使用全转录组基因表达谱分析、串联亲和纯化和质谱分析- 基于蛋白质组学方法，我们发现 p16 缺陷的黑色素瘤细胞显示出 Unfolded 的激活蛋白质反应 (UPR) 途径，并且 CDC7 可能通过以下方式维持 UPR 途径的活性转录因子HOXA1促进黑色素瘤生长和转移。因此，我们将确定 CDC7 如何通过 HOXA1 在 p16 缺陷的黑色素瘤细胞中调节 UPR 通路。此外，我们将测试 CDC7缺失诱导的p16缺陷黑色素瘤细胞的肿瘤生长和转移抑制是否可以通过恢复 HOXA1 表达和 UPR 通路活性来挽救。总的来说，我们期望发现一个新型 CDC7 驱动的可药物遗传脆弱性途径，可针对有效的个性化治疗 p16 缺陷黑色素瘤的治疗，包括高度侵袭性转移性黑色素瘤。