Breast Cancer Prevention by Ayurvedic Medicine Constituents

阿育吠陀医学成分预防乳腺癌

• 批准号: 8996137
• 负责人: Shivendra Singh
• 金额: $ 34.65万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-12-04 至 2019-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8996137
• 关键词: Accounting; Adverse effects; Affect; Alanine; Animal Model; Apoptosis; Apoptotic; Archives; Aromatase Inhibitors; Ashwagandha; Ayurvedic Medicine; Biochemical; Breast; Breast Cancer Cell; Breast Cancer Prevention; Breast Epithelial Cells; Cancer Etiology; Cancerous; Cell Death; Cell Membrane Permeability; Cells; Cessation of life; Chemoprevention; Chemopreventive Agent; Clinical; Clinical Trials Design; Cysteine; Development; Diet; Disease; Dose; Down-Regulation; Electron Transport; Electron Transport Complex III; Estrogen Receptors; Estrogen receptor negative; Estrogen receptor positive; Event; Evolution; Family; Fracture; Freezing; Funding; Growth; Health; Histone H3; Hormones; Human; Impairment; Incidence; MCF7 cell; Malignant Neoplasms; Mammary Neoplasms; Mammary gland; Mediating; Medicinal Plants; Methylnitrosourea; Microtubules; Mitochondria; Mitotic; Modeling; Modification; Molecular; Morbidity - disease rate; Mouse Mammary Tumor Virus; Neoadjuvant Therapy; PTTG1 gene; Pharmacology; Phytochemical; Plants; Post-Translational Modification Site; Post-Translational Protein Processing; Postmenopause; Prevention; Preventive Intervention; Proteomics; Publications; Publishing; Rattus; Reactive Oxygen Species; Regimen; Regulation; Research; Research Project Grants; Resistance; Risk; Risk Factors; Role; SKBR3; SUM-159 Breast Cancer Cell Line; Safety; Selective Estrogen Receptor Modulators; Technology; Testing; Toxic effect; Transgenic Mice; Tubulin; Tumor Tissue; United States; Up-Regulation; Validation; Woman; Work; base; biomarker-driven; cancer cell; cardiovascular risk factor; clinically relevant; cost; design; expectation; high risk; in vivo; innovation; knock-down; malignant breast neoplasm; mitochondrial membrane; mortality; mouse model; mutant; novel; overexpression; personalized medicine; polymerization; predictive marker; premature; prevent; receptor expression; response; small molecule; success; targeted treatment; taxane; therapeutic target; triple-negative invasive breast carcinoma; tumor

DESCRIPTION (provided by applicant): Background: Chemoprevention of breast cancer, which continues to be a leading cause of cancer-related mortality among women globally, is feasible as exemplified by clinical success of selective estrogen receptor (ER) modulators (SERM) and aromatase inhibitors. However, the SERMs are ineffective against ER- negative breast cancers and have serious side effects. The aromatase inhibitors appear promising for prevention of breast cancer in high-risk postmenopausal women. Even though the safety of aromatase inhibitors in a preventative setting is still under study, their use raises concerns for increased risk of cardiovascular events and bone fracture. Therefore, a non-toxic preventive intervention effective against both ER-positive and ER-negative breast cancers is highly desirable. In the previous funding period, we demonstrated that a small-molecule (withaferin A; hereafter abbreviated as WA) derived from an Ayurvedic medicine plant (Withania somnifera) not only prevents ER-negative mammary cancer development in a clinically-relevant transgenic mouse model (MMTV-neu) without any toxicity, but also inhibits ER- � expression and activity at pharmacologic doses in cultured human breast cancer cells. At the molecular level, we have identified novel mechanistic targets of WA, including tubulin (downregulation of �- and �- tubulin as well as covalent modification of Cys-303 of � -tubulin in MCF-7 cells) and complex III of the mitochondrial electron transport chain (activity inhibition), potentially contributing to its chemopreventive activity by eliciting mitotic arrest and reactive oxygen species (ROS)-dependent apoptotic cell death. These effects of WA are observed in mammary cancer cells representing major subtypes, including ER-positive, ER-negative, and triple-negative breast cancer. More importantly, normal mammary epithelial cells are significantly more resistant to tubulin and complex III targeting by WA compared with breast cancer cells. Despite these exciting mechanistic findings, the functional significance of down-regulation or Cys-303 modification of tubulin in growth arrest by WA is still unclear. Likewise, the molecular basis for WA-mediated inhibition of complex III activity remains elusive. Hypothesis: The present renewal application logically builds upon these novel and largely published observations to test an exciting hypothesis that WA administration prevents both ER-positive and ER-negative (already shown in the previous funding period) breast cancers in relevant animal models in association with post-translational modifications of tubulin(s) and complex III subunit(s) leading to mitotic arrest and ultimately apoptotic cell death selectively in cancerous cells. Specific Aims: The proposed research utilizes relevant cellular and in vivo animal models of breast cancer and cutting-edge technologies to: (1) determine the efficacy of dietary WA administration for prevention of ER-positive mammary cancer in a well-established rat model of chemically-induced (N-methyl- N-nitrosourea) breast cancer; (2) study the functional significance of downregulation and post-translational modification of tubulins in mitotic arrest by WA; and (3) determine the molecular mechanism by which WA inhibits complex III activity. Translational Impact: The progress in the previous funding period exceeded our own expectations as evidenced by completion and publication of the proposed work as well as identification of novel mechanistic targets of WA. The translational impact of the studies proposed in this renewal application is ultimately realized by: (a) rational design of a pilot biomarker-driven trial in a neoadjuvant window setting, which is beyond the scope of this application because clinical trial design without a full appreciation of the molecular pharmacology of WA is premature; (b) identification of mechanistic biomarker(s) predictive of WA exposure, and possibly response, which is critical for its clinical development because primary cancer incidence is too demanding of an end point; and (c) identification of a non-toxic regimen for targeting tubulin/microtubule network selectively in cancer cells as currently available anti-mitotics (e.g., taxanes) have side effects. In this era of targeted therapies and personalized medicine, the tubulin/microtubule network still remains an attractive therapeutic target for breast and other cancers.

背景：乳腺癌的化学预防仍然是全球女性癌症相关死亡的主要原因，选择性雌激素受体调节剂(SERM)和芳香酶抑制剂的临床成功证明了这一点是可行的。然而，SERM对ER阴性的乳腺癌无效，并有严重的副作用。芳香酶抑制剂似乎有希望预防高危绝经后妇女的乳腺癌。尽管芳香酶抑制剂在预防环境下的安全性仍在研究中，但它们的使用引发了人们对心血管事件和骨折风险增加的担忧。因此，一种对ER阳性和ER阴性乳腺癌都有效的无毒预防性干预是非常可取的。在之前的资助期间，我们证明了来自阿育吠陀药用植物的小分子(以下简称WA)不仅可以在没有任何毒性的临床相关转基因小鼠模型(MMTV-NEU)中阻止ER阴性乳腺癌的发展，而且在药物剂量下也可以抑制培养的人乳腺癌细胞中ER-�的表达和活性。在分子水平上，我们已经确定了WA的新的机制靶点，包括微管蛋白(下调�-和�-微管蛋白以及�-微管蛋白的半胱氨酸-303Cys-303在MCF-7细胞中的共价修饰)和线粒体电子传递链复合体III(活性抑制)，通过诱导有丝分裂停滞和活性氧(ROS)依赖的细胞死亡来潜在地参与其化学预防活性。这些作用在代表主要亚型的乳腺癌细胞中观察到，包括ER阳性、ER阴性和三阴性乳腺癌。更重要的是，与乳腺癌细胞相比，正常乳腺上皮细胞对微管蛋白和WA靶向的复合体III的抵抗力明显更强。尽管有这些令人兴奋的机制发现，下调微管蛋白或Cys-303修饰微管蛋白在WA抑制生长中的功能意义仍然不清楚。同样，WA介导的抑制复合体III活性的分子基础仍然难以捉摸。假设：目前的更新应用逻辑建立在这些新颖的和大量发表的观察结果的基础上，以测试一个令人兴奋的假说，即Wa给药可以预防相关动物模型中ER阳性和ER阴性的乳腺癌(已在上一个资助期显示)，并与微管蛋白(S)和复合体III亚单位(S)的翻译后修饰相关，从而选择性地导致癌细胞中有丝分裂停止，最终导致细胞凋亡。具体目的：拟议的研究利用相关的乳腺癌细胞和体内动物模型以及尖端技术来：(1)确定饮食中给予水杨酸对已建立的化学诱导(N-甲基-N-亚硝胺)乳腺癌大鼠模型中ER阳性乳腺癌的预防效果；(2)研究微管蛋白下调和翻译后修饰在水杨酸抑制有丝分裂中的功能意义；(3)确定水杨酸抑制复合体III活性的分子机制。翻译影响：前一个供资期间的进展超出了我们自己的预期，这体现在拟议工作的完成和出版以及确定了西澳的新机制目标。在这一更新申请中提出的研究的翻译影响最终通过：(A)在新的辅助窗口设置下合理地设计试点生物标记物驱动的试验，这超出了本申请的范围，因为在没有充分了解WA的分子药理学的情况下进行临床试验设计是过早的；(B)确定预测WA暴露和可能的反应的机械性生物标记物(S)，这对其临床发展至关重要，因为原发癌症发病率对终点要求太高；以及(C)由于目前可用的抗有丝分裂药物(例如紫杉烷)有副作用，因此确定一种用于在癌细胞中选择性地靶向微管/微管网络的无毒方案。在这个靶向治疗和个性化医学的时代，微管/微管网络仍然是乳腺癌和其他癌症的有吸引力的治疗靶点。