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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：
10294457
负责人：
Romi Gupta
金额：
$ 32.57万
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/10294457
关键词：
American Amidohydrolases Attenuated Automobile Driving Basic Science Breast Cancer Cell Breast Cancer Patient Breast Cancer cell line Breast Cancer therapy Bromodomain 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 Injections Mammary Neoplasms Measures Mediating Metabolic Metabolic Pathway Modeling Monitor Mus Mutate Mutation National Cancer Institute Neoplasm Metastasis Normal Cell Pathway interactions Patients Pharmacology Precision therapeutics Prognosis Proteins Public Health Role SLC2A1 gene Sampling Series Starvation TP53 gene Testing The Cancer Genome Atlas Therapeutic TimeLine Tumor stage Tumor-Derived Up-Regulation Woman Xenograft procedure adenylate kinase base cancer cell cancer subtypes cancer therapy chemotherapy effective therapy experimental study humanized mouse improved in vivo inhibitor/antagonist innovation knock-down malignant breast neoplasm mammary metabolome metabolomics mouse model mutant neoplastic cell non-genetic novel therapeutic intervention patient derived xenograft model prevent response small hairpin RNA small molecule small molecule inhibitor targeted treatment 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.

项目摘要癌症基因组图谱（TCGA）研究已经确定p53是乳腺癌中最常见的突变基因。癌特别是，80%的三阴性乳腺癌（TNBC）含有p53突变。但没有可用于治疗p53突变型TNBC的特异性疗法。癌细胞有不同的代谢需要，这一发现刺激了小分子抑制剂的发展，以靶向代谢癌细胞生存所需的酶。值得注意的是，尽管有证据表明代谢途径失调在乳腺癌生长中的作用，p53突变体的代谢依赖性 TNBC生长和转移尚未得到全面鉴定。因此，为了具体确定 p53突变型TNBCs的代谢依赖性，我们进行了一个创新的和无偏见的大规模体内短发夹RNA（shRNA）通过靶向2000个具有已知代谢功能基因进行筛选。有了这个屏幕，我们鉴定出N-酰基鞘氨醇酰胺水解酶1（ASAH 1）是肿瘤形成所必需的酶， p53突变型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， ASAH 1具有有效的抗p53突变TNBC活性，从而表明ASAH 1是TNBC的潜在药物靶标。 p53突变型TNBC的治疗。该提议的中心假设是p53突变型TNBC细胞它们的生存依赖于ASAH 1，因此，ASAH 1抑制选择性地根除p53突变型TNBC。我们的总体目标是确定ASAH 1在驱动p53突变型TNBC肿瘤生长中的作用，转移，了解p53突变型TNBC对ASAH 1依赖性的机制，评估小分子ASAH 1抑制剂治疗p53突变型TNBC的翻译潜力。在目标1中，我们将通过一系列的研究来确定ASAH 1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，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。在Aim中 2，我们将测试我们的假设，即p53抑制ASAH 1的活性，通过隔离这种蛋白质在细胞核中。此外，基于我们新的初步结果，我们将确认ASAH 1的缺失是否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激酶途径活化细胞膜上的葡萄糖转运蛋白GLUT 1。在目标3中，我们将评估 P53突变体的免疫受损和免疫活性人源化小鼠模型中的ASAH 1抑制剂 TNBC，单独或基于我们的初步发现，与BET结构域抑制剂组合。总的来说，我们预测，本申请中提出的实验结果将确立ASAH 1作为一种重要的 p53-突变TNBC细胞固有的脆弱性，阐明了p53-突变TNBC细胞依赖性的机制。本发明的目的是研究突变型TNBC对ASAH 1活性的影响，并评估用于治疗p53突变型TNBC的新的治疗方法。