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

• 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
• 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; 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

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 癌症基因组图集（TCGA）研究已确定p53是乳房中最常见的突变基因 癌症。特别是，有80％的三阴性乳腺癌（TNBC）携带p53突变。但是，没有 可用于治疗p53突变TNBC的特定疗法。癌细胞比较有不同的代谢需求 对于正常细胞，这种观察刺激了小分子抑制剂的发展以靶向代谢 癌细胞特别需要的酶才能生存。值得注意的是，尽管有证据支持 代谢途径放松管制在乳腺癌生长中的作用，p53突变的代谢依赖性 TNBC的生长和转移尚未全面鉴定。因此，专门确定 p53突变型TNBC的代谢依赖性，我们进行了创新且无偏的大规模体内 通过靶向具有已知代谢功能的2000个基因，短发RNA（SHRNA）筛选。在此屏幕上， 我们将N-酰基肾上腺素酶1（ASAH1）确定为肿瘤形成所必需的酶 p53突变TNBC的能力和转移活性。此外，我们确定了两个小分子抑制剂 ASAH1具有有效的抗P53突变TNBC活性，从而表明ASAH1是潜在的药物靶标 P53突变TNBC的处理。该提议的中心假设是p53-突出TNBC细胞 依靠ASAH1的生存，因此，ASAH1抑制选择性地消除了p53突变的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-突动剂TNBC对ASAH1的依赖性的机制，并 评估小分子ASAH1抑制剂用于治疗p53突变TNBC的转化潜力。在AIM 1中， 我们将使用一系列ASAH1在p53突变的TNBC肿瘤生长和转移中的作用 互补的，最先进的小鼠模型，这些模型概括了TNBC增长的特征和 转移。这些包括具有人类免疫系统的高度创新的人性化小鼠模型。目标 2，我们将测试我们的假设，即p53通过隔离核中的该蛋白质来抑制ASAH1活性。 此外，根据我们的新初步结果，我们将确认ASAH1的损失是否激活葡萄糖 饥饿反应，导致通过神经酰胺介导的表达降低的AMP激酶途径激活 细胞膜上的葡萄糖转运蛋白glut1。在AIM 3中，我们将评估的翻译潜力 ASAH1抑制剂p53突变物的免疫功能低下和免疫能力的人型小鼠模型 TNBC是单独的，或基于我们的初步发现，结合了BET结构域抑制剂。共同 我们预测，本应用程序中提出的实验结果将确立ASAH1为重要的 p53突变TNBC细胞固有的脆弱性，阐明了p53-依赖性的机制 在ASAH1活性上突变TNBC，并评估一种用于治疗p53突变TNBC的新型治疗方法。