Targeted Inhibition in Triple Negative Breast Cancer

三阴性乳腺癌的靶向抑制

• 批准号: 9750252
• 负责人: Nicolas Nassar
• 金额: $ 20.75万
• 依托单位: CINCINNATI CHILDRENS HOSP MED CTR
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-07-24 至 2020-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9750252
• 关键词: Acute; Binding; Breast Cancer Cell; Breast Cancer Patient; Breast Cancer Treatment; Breast Cancer therapy; Cancer Cell Growth; Cell Culture Techniques; Cell Survival; Cells; Cessation of life; Chronic; Clinic; Clinical; Clinical Trials; Data; Disease; Dose; Drug Kinetics; Emergency Department patient; Epidermal Growth Factor Receptor; Epithelial Cells; Estrogen receptor positive; FRAP1 gene; Genetic; Goals; Growth; Health; Homeostasis; Individual; Intervention; Lead; Molecular Target; Mus; Myeloid Leukemia; Neoplasm Metastasis; Normal Cell; Oncogenic; Operative Surgical Procedures; Pathway interactions; Pharmacogenetics; Phase I Clinical Trials; Phenotype; Phosphoric Monoester Hydrolases; Phosphorylation; Preclinical Testing; Protein Tyrosine Kinase; Protein Tyrosine Phosphatase; Proteomics; Proto-Oncogene Proteins c-akt; Radiation; Relapse; Research Personnel; Research Proposals; Resistance; Serum; Signal Transduction; Signaling Protein; Site-Directed Mutagenesis; Structure; Testing; Therapeutic; Time; Toxic effect; Ubiquitination; Woman; Xenograft procedure; analog; base; biophysical techniques; cancer subtypes; cancer type; cell growth; cellular targeting; chemotherapy; dosage; efficacy testing; hormone therapy; human model; in vivo; inhibitor/antagonist; leukemia; malignant breast neoplasm; mortality; mouse model; novel; novel therapeutics; outcome forecast; overexpression; phosphatase inhibitor; preclinical safety; proto-oncogene protein c-cbl; side effect; small hairpin RNA; small molecule; small molecule inhibitor; small molecule therapeutics; targeted treatment; tool; triple-negative invasive breast carcinoma; tumor; tumor growth; tumor xenograft

ABSTRACT We have identified a small molecule lead that at low doses decreases the proliferation of triple negative breast cancer (TNBC) cells while sparing normal epithelial cells. Mechanistically, our lead targets the phosphatase activity of Sts-1, a novel target that is overexpressed in TNBC and whose high expression confers poor overall survival in ER-(-) but not in ER-(+) patients. Our preliminary data show that inhibiting Sts-1 is a potent mechanism of treatment, as tumors induced by HCC1937 cell xenografts in mice carrying a Sts-1 targeting shRNA grew significantly smaller than tumors from control cells with non-targeting shRNA. In cell culture, decreasing Sts-1 expression decreased proliferation of TNBC but not of control cells. Given these activities, the goal of this research proposal is to validate the mechanism of action and to test the efficacy of our lead in an in vivo xenograft mouse model of TNBC. We will focus on TNBC because it is among the most aggressive of breast cancer subtype, with a high propensity for metastasis, poor prognosis, and short time to relapse and death. TNBC represents a clinical challenge because it does not benefit from hormonal therapies or other targeted treatments. Chemotherapy, with its adverse side effects, remains the cornerstone for TNBC treatment and chemo- agents are often administered individually or in combination; surgery and radiation are also commonly used. Our long-term goal is to develop small molecule inhibitors of the phosphatase activity of Sts-1 into a novel TNBC therapy. Our working hypothesis is that small molecule inhibitors of Sts-1 will restore c-Cbl's ability to degrade its targets (e.g. EGFR, c-Met) by ubiquitination - thereby inhibiting TNBC growth. To validate the mechanism of action of H9 and determine its potency in vivo, Aim 1 will test the effect of H9 on the activity of the c-Cbl/EGFR, c-Cbl/c-Met, and mTOR/S6K axes under acute and chronic growth conditions, investigate the cellular pathways modulated by compound H9 and compare them to Sts-1 silencing using unbiased approaches, and examine and validate binding of H9 to Sts-1 phosphatase domain using structural and biophysical techniques and site-directed mutagenesis. Aim 2 will test the ability of compound H9 to inhibit growth of TNBC tumors in vivo. We will Initiate toxicity and pharmacokinetic studies to determine compound stability and optimal dosage to achieve functional serum concentrations of compound H9 and test the ability of H9 to inhibit growth of TNBC xenograft tumors in vivo.

摘要 我们已经确定了一种小分子铅，在低剂量下可以减少三阴性细胞的增殖， 乳腺癌（TNBC）细胞，而不包括正常上皮细胞。从机制上讲，我们的线索目标是 Sts-1是一种在TNBC中过表达的新靶点，其高表达 ER-（-）患者的总生存率较低，但ER-（+）患者的总生存率较低。初步数据显示， 抑制Sts-1是一种有效的治疗机制，因为HCC 1937细胞异种移植物诱导的肿瘤在 携带Sts-1靶向shRNA的小鼠生长的肿瘤明显小于对照细胞， 非靶向shRNA。在细胞培养中，降低Sts-1表达降低了TNBC的增殖， 而不是控制细胞。鉴于这些活动，本研究提案的目标是验证 本发明的目的是为了研究其作用机制，并测试我们的先导化合物在TNBC的体内异种移植小鼠模型中的功效。 我们将关注TNBC，因为它是最具侵袭性的乳腺癌亚型之一， 转移倾向、预后差、复发和死亡时间短。TNBC代表一个 临床挑战，因为它不受益于激素疗法或其他靶向治疗。 化疗及其不良副作用仍然是TNBC治疗和化疗的基石。 药剂通常单独或组合施用;手术和放射也通常 采用我们的长期目标是开发Sts-1磷酸酶活性的小分子抑制剂 一种新的TNBC疗法 我们的工作假设是，Sts-1的小分子抑制剂将恢复c-Cbl的降解能力， 通过泛素化作用抑制其靶点（例如EGFR、c-Met），从而抑制TNBC生长。验证 为了研究H9的作用机制并确定其在体内的效力，目的1将测试H9对 急性和慢性生长条件下c-Cbl/EGFR、c-Cbl/c-Met和mTOR/S6 K轴的活性 条件下，研究由化合物H9调节的细胞途径，并将它们与Sts-1进行比较 使用无偏的方法沉默，并检查和验证H9与Sts-1磷酸酶的结合 结构域使用结构和生物物理技术和定点诱变。目标2将测试 化合物H9在体内抑制TNBC肿瘤生长的能力。我们将启动毒性， 药代动力学研究，以确定化合物的稳定性和实现功能的最佳剂量。 测定化合物H9的血清浓度并测试H9抑制TNBC异种移植物生长的能力 体内肿瘤