The Role of FoxQ1 in Breast Cancer Chemoprevention by Allium Constituents - R01CA219180

摘要 背景：乳腺癌仍然是全球女性癌症相关死亡的主要原因。 尽管我们对人类免疫缺陷病毒的生物学、风险因素和基因组图景的了解日益广泛 疾病。化学预防是降低乳房发病率和死亡率的有效策略。 癌症。临床上对选择性雌激素的持续兴趣证明了这种方法的可行性和前景。 受体(ER)调节剂(例如，他莫昔芬)，以及最近的芳香酶抑制剂(例如，西美坦) 管腔型乳腺癌的化学预防。不幸的是，这些干预措施有副作用，而且缺乏 对雌激素受体阴性的乳腺癌亚型(例如，基底细胞样乳腺癌)的活性。因此，一个安全和 针对不同亚型乳腺癌的廉价化学预防干预仍然有效 令人向往。这项临床前研究项目的首要目标是获取活体证据 一种前景看好的饲料植物化学物质(二烯丙基三硫化物；DATS)的化学预防效果 使用啮齿动物模型的蔬菜(例如大蒜)表现出与基类和 管腔型人类乳腺癌。与人类相关的动物体内化学预防效果的证据 模型是启动DATS临床试验的先决条件，这在以前的第一个人类中是被很好地耐受的 间歇给药方案的干预性研究。发表的作品，包括我们实验室的作品， 已经证明了DATS对基底样型和管腔型人乳腺癌细胞株的活性 体外，以及它们的异种移植和体内的肿瘤干细胞(BCSC)群体。流行病学研究还发现 研究表明，饮食中大蒜和洋葱的摄入量与乳腺癌风险呈负相关。 这项提议的机械方面是非常新颖的，并且围绕着一个仍然鲜为人知的 转录因子，叉头盒Q1(FoxQ1)。受益于从TCGA获得RNA-Seq数据 数据库，并通过靶向基因表达谱，我们已经确定了FoxQ1的新靶点，包括 腊肠同系物1(DACH1)和单羧酸转运蛋白1(MCT1)。DACH1是细胞的命运 决定因子和肿瘤抑制因子，而MCT1在代谢调节中具有致癌作用 在癌细胞中重新编程。我们最近发表的研究表明，FoxQ1是一个直接转录的 DACH1的抑制子，因此FoxQ1的表达与DACH1的表达呈负相关 人类乳腺癌(TCGA)另一方面，FoxQ1的表达与FoxQ1表达呈正相关 MCT1.我们还发现DATS处理抑制了FoxQ1和MCT1的表达，但诱导了DACH1 体外培养乳腺癌细胞中的蛋白质。然而，这些细胞发现在体内的相关性仍然不清楚。 同样，FoxQ1/MCT1轴在乳腺癌进展中的作用及其在癌症中的潜在作用 DATS的化学预防作用尚不清楚。因此，机械论的研究在逻辑上集中在 FoxQ1及其下游新靶点(DACH1和MCT1)在乳腺癌化学预防中的作用 DATS。假设：我们假设口服DATS可阻止基底样和 人相关啮齿动物模型中管腔型乳腺癌的研究 FoxQ1(BCSC的自我更新和代谢从糖酵解转换到氧化磷酸化)。 具体目的：(1)确定DATS化学预防基底样癌的体内疗效 使用转基因小鼠模型，以及(B)使用大鼠模型进行管腔型乳腺癌；(2)确定 DATS对人基底细胞样型和腔型乳腺癌BCSC抑制作用中FoxQ1/DACH1轴的表达 确定FoxQ1/MCT1轴在代谢重编程中的作用以及DATS对其的抑制作用 使用细胞模型和代表人类基底样亚型和管腔亚型的乳腺肿瘤组织阵列。 来自对照和DATS处理的小鼠和大鼠(AIM 1)的肿瘤组织将被用来确定体内的 FoxQ1/DACH1和FoxQ1/MCT1轴在DATS化疗预防乳腺癌中的意义 意义：化学预防有效性的临床前证据是证明DATS临床试验的合理性的关键 乳腺癌患者。第一个特定目标直接测试化学预防类基底细胞和 使用特征良好的啮齿动物模型通过DATS给药的管腔型乳腺癌。建议数 机制研究可能区分药效学生物标志物(S)，可能在未来的临床研究中有用 用DATS。最后，进一步了解FoxQ1在乳腺癌中的作用可能最终导致新的 治疗这种异质疾病的方法。

ABSTRACT Background: Breast cancer remains a leading cause of cancer-related deaths among women worldwide despite our increasingly broader understanding of the biology, risk factors, and genomic landscape of the disease. Chemoprevention signifies a meaningful strategy for reducing the morbidity and mortality from breast cancer. Feasibility and promise of this approach is illustrated by continued clinical interest in selective estrogen receptor (ER) modulators (e.g., tamoxifen), and more recently, aromatase inhibitors (e.g., exemestane) for chemoprevention of luminal-type breast cancers. Unfortunately, these interventions have side effects and lack activity against ER-negative subtypes of breast cancer (e.g., basal-like breast cancer). Therefore, a safe and inexpensive chemopreventive intervention efficacious against different subtypes of breast cancer is still desirable. The overarching goal of this preclinical research project is to acquire in vivo evidence for chemopreventive efficacy of a highly promising dietary phytochemical (diallyl trisulfide; DATS) from Allium vegetables (e.g., garlic) using rodent models exhibiting significant molecular overlap with basal-like and luminal-type human breast cancers. In vivo evidence of chemopreventive efficacy in human-relevant animal models is a prerequisite for initiation of clinical trials of DATS, which was well-tolerated in a prior first-in-human interventional study with intermittent dosing schedule. Published work, including that from our laboratory, already demonstrates activity of DATS against basal-like and luminal-type human breast cancer cell lines in vitro, and their xenografts and cancer stem cell (bCSC) populations in vivo. Epidemiological studies have also suggested an inverse association between dietary intake of garlic and onions and the risk of breast cancer. The mechanistic aspects of this proposal are exceedingly novel and revolve around a still poorly understood transcription factor, Forkhead box Q1 (FoxQ1). Benefitted by access to the RNA-Seq data from TCGA database and through targeted gene expression profiling, we have identified novel targets of FoxQ1, including Dachshund homolog 1 (DACH1) and monocarboxylate transporter 1 (MCT1). DACH1 is a cell fate determination factor and tumor suppressor, whereas MCT1 has an oncogenic role in regulation of metabolic reprogramming in cancer cells. Our recently published work indicates that FoxQ1 is a direct transcriptional repressor of DACH1, and consequently FoxQ1 expression is inversely associated with that of DACH1 in human breast cancers (TCGA). On the other hand, FoxQ1 expression is positively associated with that of MCT1. We also found that DATS treatment suppresses FoxQ1 and MCT1 expression but induces DACH1 protein in breast cancer cells in vitro. However, the in vivo relevance of these cellular findings is still unclear. Likewise, the contribution of FoxQ1/MCT1 axis to breast cancer progression and its potential role in cancer chemoprevention by DATS are yet to be elucidated. Therefore, the mechanistic studies logically focus on the role of FoxQ1 and its novel downstream targets (DACH1 and MCT1) in breast cancer chemoprevention by DATS. Hypothesis: We hypothesize that oral administration of DATS prevents development of basal-like and luminal-type breast cancer in human-relevant rodent models by suppressing oncogenic processes regulated by FoxQ1 (self-renewal of bCSC and metabolic switch from glycolysis to oxidative phosphorylation). Specific Aims: (1) Determine the in vivo efficacy of DATS for chemoprevention of: (a) basal-like breast cancer using a transgenic mouse model, and (b) luminal-type breast cancer using a rat model; (2) Determine the role of FoxQ1/DACH1 axis in bCSC inhibition by DATS using human basal-like and luminal-type breast cancer cells; and (3) Determine the role of FoxQ1/MCT1 axis in metabolic reprogramming and its inhibition by DATS using cellular models and breast tumor tissue arrays representative of human basal-like and luminal subtypes. The tumor tissues from control and DATS-treated mice and rats (aim 1) will be used to determine the in vivo significance of FoxQ1/DACH1 and FoxQ1/MCT1 axes in breast cancer chemoprevention by DATS. Significance: Preclinical evidence of chemopreventive efficacy is essential to justify clinical trial of DATS in breast cancer patients. The first specific aim directly tests the possibility of chemoprevention of basal-like and luminal-type breast cancer by DATS administration using well-characterized rodent models. The proposed mechanistic studies may distinguish pharmacodynamic biomarker(s) potentially useful in future clinical studies with DATS. Finally, further understanding of the role of FoxQ1 in breast cancer may ultimately lead to novel therapies for this heterogeneous disease.