PRECISION METABOLIC THERAPY OF p53 MUTANT TRIPLE NEGATIVE BREAST CANCERS

p53 突变三阴性乳腺癌的精准代谢治疗

基本信息

批准号：
10621315
负责人：
Romi Gupta
金额：
$ 31.92万
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：
2021
资助国家：
美国
起止时间：
2021-07-20 至 2026-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10621315
关键词：
American Amidohydrolases Attenuated Automobile Driving Basic Science Breast Cancer Cell Breast Cancer Patient Breast Cancer cell line Breast Cancer therapy Bromodomain Bromodomains and extra-terminal domain inhibitor CRISPR/Cas technology Cell Culture Techniques Cell Nucleus Cell Survival Cell membrane Ceramidase Ceramides Characteristics Clinical Combined Modality Therapy Data Dependence Development Diagnosis Disease Doxycycline Drug Targeting ERBB2 gene Enzymes Estrogen receptor positive Fatty acid glycerol esters Female Genes Genetic Glucose Glucose Transporter Growth Guide RNA Human Immune Evasion Immune system Immunocompetent Immunocompromised Host Immunodeficient Mouse Immunologic Deficiency Syndromes Injections Mammary Neoplasms Measures Mediating Metabolic Metabolic Pathway Modeling Monitor Mus Mutate Mutation National Cancer Institute Neoplasm Metastasis Normal Cell Pathway interactions Patients Precision therapeutics Prognosis Proteins Public Health Repression Role SLC2A1 gene Sampling Series Starvation TP53 gene Testing The Cancer Genome Atlas Therapeutic Tumor Promotion Tumor stage Up-Regulation Woman Xenograft procedure adenylate kinase cancer cell cancer subtypes cancer therapy chemotherapy effective therapy experimental study humanized mouse improved in vivo inhibitor innovation knock-down malignant breast neoplasm mammary metabolome metabolomics mouse model mutant neoplastic cell non-genetic novel therapeutic intervention patient derived xenograft model pharmacologic prevent response small hairpin RNA small molecule small molecule inhibitor targeted treatment timeline translational potential triple-negative invasive breast carcinoma tumor tumor growth tumor heterogeneity tumor microenvironment tumor progression

项目摘要

PROJECT SUMMARY The Cancer Genome Atlas (TCGA) studies have identified p53 as the most frequently mutated gene in breast cancer. In particular, 80% of triple-negative breast cancers (TNBCs) harbor p53 mutations. However, there is no specific therapy available for treating p53-mutant TNBC. Cancer cells have distinct metabolic needs compared to normal cells, and this observation has spurred the development of small molecule inhibitors to target metabolic enzymes that are specifically needed by cancer cells for survival. Notably, although there is evidence to support a role for metabolic pathway deregulation in breast cancer growth, the metabolic dependencies of p53-mutant TNBC growth and metastasis have not been comprehensively identified. Therefore, to specifically identify the metabolic dependencies of p53-mutant TNBCs, we performed an innovative and unbiased large-scale in vivo short-hairpin RNA (shRNA) screen by targeting 2000 genes with known metabolic functions. With this screen, we identified N-acylsphingosine amidohydrolase 1 (ASAH1) as an enzyme that is necessary for tumor-forming ability and metastatic activity of p53-mutant TNBCs. Additionally, we identified two small molecule inhibitors of ASAH1 with potent anti-p53-mutant TNBC activity, thereby indicating that ASAH1 is a potential drug target for the treatment of p53-mutant TNBCs. The central hypothesis of this proposal is that p53-mutant TNBC cells depend upon ASAH1 for their survival, and thus, ASAH1 inhibition selectively eradicates p53-mutant TNBCs. Our overall objectives are to determine the role of ASAH1 in driving p53-mutant TNBC tumor growth and metastasis, understand the mechanism underlying the dependency of p53-mutant TNBCs on ASAH1, and evaluate the translational potential of small molecule ASAH1 inhibitors for treating p53-mutant TNBC. In Aim 1, we will establish the role of ASAH1 in p53-mutant TNBC tumor growth and metastasis using a series of complementary, state-of-the-art mouse models that recapitulate characteristic features of TNBC growth and metastasis. These include a highly innovative humanized mouse model with a human immune system. In Aim 2, we will test our hypotheses that p53 represses ASAH1 activity by sequestering this protein in the nucleus. Additionally, based on our new preliminary results, we will confirm whether loss of ASAH1 activates the glucose starvation response, leading to AMP kinase pathway activation via a ceramide-mediated reduction in expression of the glucose transporter GLUT1 on the cell membrane. In Aim 3, we will evaluate the translational potential of ASAH1 inhibitors in immunocompromised and immunocompetent humanized mouse models of p53-mutant TNBC, either alone, or based on our preliminary findings, in combination with BET domain inhibitors. Collectively, we predict that the results of the experiments proposed in this application will establish ASAH1 as an important vulnerability inherent to p53-mutant TNBC cells, elucidate the mechanism underlying the dependency of p53- mutant TNBCs on ASAH1 activity, and evaluate a novel therapeutic approach for treatment of p53-mutant TNBC.

项目概要癌症基因组图谱 (TCGA) 研究已确定 p53 是乳腺癌中最常见的突变基因癌症。特别是，80% 的三阴性乳腺癌 (TNBC) 带有 p53 突变。然而，没有可用于治疗 p53 突变 TNBC 的特异性疗法。相比之下，癌细胞具有不同的代谢需求对正常细胞的影响，这一观察刺激了针对代谢的小分子抑制剂的开发癌细胞生存所特别需要的酶。值得注意的是，尽管有证据支持 p53突变体的代谢依赖性在乳腺癌生长中代谢途径失调的作用 TNBC 的生长和转移尚未得到全面鉴定。因此，要具体识别为了研究 p53 突变 TNBC 的代谢依赖性，我们进行了一次创新且公正的大规模体内研究通过针对 2000 个具有已知代谢功能的基因进行短发夹 RNA (shRNA) 筛选。有了这个屏幕，我们确定 N-酰基鞘氨醇酰胺水解酶 1 (ASAH1) 是肿瘤形成所必需的酶 p53 突变 TNBC 的能力和转移活性。此外，我们还发现了两种小分子抑制剂 ASAH1 具有有效的抗 p53 突变 TNBC 活性，从而表明 ASAH1 是潜在的药物靶点 p53突变TNBC的治疗。该提案的中心假设是 p53 突变 TNBC 细胞它们的生存依赖于 ASAH1，因此，ASAH1 抑制选择性地根除 p53 突变 TNBC。我们的总体目标是确定 ASAH1 在驱动 p53 突变 TNBC 肿瘤生长和转移，了解 p53 突变 TNBC 对 ASAH1 依赖性的机制，以及评估小分子 ASAH1 抑制剂治疗 p53 突变 TNBC 的转化潜力。在目标 1 中，我们将使用一系列的方法来确定 ASAH1 在 p53 突变型 TNBC 肿瘤生长和转移中的作用互补的、最先进的小鼠模型，概括了 TNBC 生长的特征和转移。其中包括具有人类免疫系统的高度创新的人源化小鼠模型。瞄准 2，我们将测试我们的假设，即 p53 通过将这种蛋白质隔离在细胞核中来抑制 ASAH1 活性。此外，根据我们新的初步结果，我们将确认 ASAH1 的缺失是否会激活葡萄糖饥饿反应，通过神经酰胺介导的表达减少导致 AMP 激酶途径激活细胞膜上的葡萄糖转运蛋白 GLUT1。在目标 3 中，我们将评估以下内容的转化潜力： p53 突变体免疫功能低下和免疫功能正常的人源化小鼠模型中的 ASAH1 抑制剂 TNBC，可以单独使用，也可以根据我们的初步发现，与 BET 结构域抑制剂联合使用。总的来说，我们预测本申请中提出的实验结果将确立 ASAH1 作为重要的 p53 突变 TNBC 细胞固有的脆弱性，阐明了 p53- 依赖性的潜在机制突变 TNBC 对 ASAH1 活性的影响，并评估治疗 p53 突变 TNBC 的新治疗方法。