Biomarkers of Sulforaphane/Broccoli Sprout Extract in Prostate Cancer

萝卜硫素/西兰花芽提取物在前列腺癌中的生物标志物

• 批准号: 10314024
• 负责人: Shivendra Singh
• 金额: $ 66.05万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-01-01 至 2023-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10314024
• 关键词: ATP Citrate (pro-S)-Lyase; Acetates; Acetyl Coenzyme A; Acetyl-CoA Carboxylase; Adenocarcinoma; Affect; American; Biochemical; Bioenergetics; Biological Availability; Biological Markers; Broccoli - dietary; Cancer Cell Growth; Cancer Etiology; Cell Proliferation; Cell model; Cells; Cessation of life; Chemoprevention; Chemopreventive Agent; Clinical; Clinical Research; Clinical Trials; Coupled; Data; Dietary Administration; Disease; Double-Blind Method; Down-Regulation; Edible Plants; Enzyme Inhibition; Enzymes; Epithelial; Exposure to; Fatty Acids; Fatty-acid synthase; Formulation; Foundations; Genetic; Genetic Transcription; Glycolysis; Goals; Human; In Vitro; Incidence; Individual; Induction of Apoptosis; Intervention; Link; Malignant Neoplasms; Malignant neoplasm of prostate; Malonyl Coenzyme A; Mediating; Medicinal Plants; Messenger RNA; Mitochondria; Molecular; Morbidity - disease rate; Mus; Neoplasms; Nonesterified Fatty Acids; Normal tissue morphology; Oral; Oral Administration; Outcome Study; Oxidoreductase; PTEN gene; Pathogenesis; Pathway interactions; Pharmacology; Phospholipids; Phytochemical; Placebo Control; Positron-Emission Tomography; Pre-Clinical Model; Prostate; Prostatectomy; Prostatic; Prostatic Intraepithelial Neoplasias; Prostatic Neoplasms; Protein Inhibition; Proteins; Publishing; Randomized; Research; Rodent Model; Role; SRE-1 binding protein; Safety; Schedule; Serum; Solid Neoplasm; Sulforaphane; Technology; Testing; Transgenic Mice; Warburg Effect; c-myc Genes; cancer cell; cancer chemoprevention; clinical development; clinically relevant; design; experimental study; fatty acid metabolism; fatty acid oxidation; human disease; in vivo; in vivo imaging; lipidomics; male; men; mortality; mouse model; overexpression; oxidation; pharmacodynamic biomarker; pilot trial; placebo group; pre-clinical; preclinical study; primary endpoint; prostate cancer cell; prostate cancer model; secondary endpoint; transgenic adenocarcinoma of mouse prostate; translational impact; translational study; tumor; uptake

ABSTRACT Scientific Premise and Hypothesis: Chemoprevention using safe and inexpensive phytochemicals from edible or medicinal plants is appealing for reducing the death and suffering from prostate cancer, which continues to be a leading cause of cancer-linked mortality among American men. A chemopreventative intervention for prostate cancer is still lacking. Increased de novo synthesis coupled with β-oxidation of fatty acids is a rather unique and targetable mechanism of human prostate cancer. A role for upregulated de novo fatty acid synthesis in pathogenesis of prostate cancer is substantiated by studies showing overexpression of mRNA/protein levels of key fatty acid synthesis enzymes, including ATP citrate lyase (ACLY), acetyl-CoA carboxylase 1 (ACC1), and/or fatty acid synthase (FASN) in early (prostatic intraepitheilial neoplasia; PIN) as well as advanced (adenocarcinoma) disease when compared to normal tissue. In addition, genetic or pharmacological suppression of ACLY, ACC1, and FASN causes inhibition of prostate cancer cell growth in vitro and in vivo. Therefore, inhibition of synthesis and/or β-oxidation of fatty acids represents a promising strategy for chemoprevention of prostate cancer. The overall goal of this bench-cage-bedside project is to determine the feasibility of fatty acid metabolism inhibition for chemoprevention of prostate cancer using sulforaphane (SFN), which is the principal bioactive phytochemical in broccoli sprout extract (BSE). The preclinical studies are conceived to test the hypothesis that prostate cancer chemoprevention by SFN and BSE in a clinically-relevant transgenic mouse model (Hi-Myc) is associated with suppression of synthesis as well as β-oxidation of fatty acids leading to inhibition of cancer cell proliferation. A pilot double-blind, randomized, and placebo-controlled window of opportunity clinical trial in men scheduled for prostatectomy is also proposed to determine whether daily oral administration of a well-characterized BSE formulation (BroccoMax®), the safety of which has already been tested clinically, for 4 weeks leads to suppression of circulating and prostate tumor levels of fatty acids. Support for the above stated hypothesis derives from our own published and preliminary unpublished findings. Specifically, we found that SFN treatment not only suppresses protein/mRNA levels of ACC1 and FASN as well as the dehydrogenases implicated in β-oxidation of fatty acids but also decreases acetyl-CoA levels in human prostate cancer cells in vitro and prostate tumors of TRAMP mice in vivo. Acetyl-CoA is the building block of de novo fatty acid synthesis but is also generated in the mitochondria upon β-oxidation of fatty acids. Specific Aims: The well-integrated specific aims of this highly-focused application are to: (1) Determine the mechanism underlying SFN-mediated inhibition of fatty acid synthesis and β-oxidation using cellular models of prostate cancer and normal prostate cells; (2) Determine whether prostate cancer chemoprevention by SFN and BSE in Hi-Myc transgenic mice is associated with inhibition of fatty acid synthesis and β-oxidation; and (3) Determine whether daily oral BroccoMax® administration for 4 weeks decreases circulating and prostate tumor levels of fatty acids through a pilot double-blind, randomized, and placebo-controlled window of opportunity trial in men scheduled for prostatectomy. Translational Impact (Significance): Despite wealth of mechanistic data, clinical development of SFN or BSE like BroccoMax® for chemoprevention of prostate cancer is contingent upon: (a) demonstration of chemopreventive efficacy in a clinically-relevant rodent model of prostate cancer; (b) identification of pharmacodynamic biomarker(s) using cellular and preclinical rodent models of prostate cancer; and (c) pilot clinical translational studies to demonstrate modulation of the same mechanistic biomarker(s) identified from the cellular/preclinical models such as suppression of fatty acid metabolism hypothesized in this application. This stepwise progression of research is essential to justify a larger clinical study with prostate cancer incidence as the primary end point. The first two specific aims of this project will not only test the possibility of prostate cancer chemoprevention by SFN and BSE in a transgenic mouse model with strong molecular overlap with the human disease but will also identify non-invasive pharmacodynamic biomarker(s) (e.g., serum levels of fatty acids) applicable to the proposed clinical trial in specific aim 3.

抽象的 科学前提和假设：使用安全且廉价的食用植物化学物质进行化学预防 或药用植物对于减少前列腺癌的死亡和痛苦具有吸引力，前列腺癌仍在继续 是美国男性癌症相关死亡的主要原因。化学预防干预 前列腺癌仍然缺乏。增加从头合成与脂肪酸的β-氧化相结合是一种相当有效的方法。 人类前列腺癌的独特且可靶向的机制。上调脂肪酸从头合成的作用 mRNA/蛋白质水平过度表达的研究证实了前列腺癌的发病机制 关键脂肪酸合成酶，包括 ATP 柠檬酸裂解酶 (ACLY)、乙酰辅酶A羧化酶 1 (ACC1)、 和/或脂肪酸合酶（FASN）在早期（前列腺上皮内瘤变；PIN）以及晚期 与正常组织相比，（腺癌）疾病。此外，遗传或药物抑制 ACLY、ACC1 和 FASN 的作用可在体外和体内抑制前列腺癌细胞的生长。所以， 抑制脂肪酸的合成和/或β-氧化代表了一种有前途的化学预防策略 前列腺癌。这个实验台-笼子-床边项目的总体目标是确定脂肪酸的可行性 使用萝卜硫素（SFN）抑制代谢以化学预防前列腺癌，这是主要的 西兰花芽提取物 (BSE) 中的生物活性植物化学物质。临床前研究旨在测试 假设SFN和BSE在临床相关转基因中对前列腺癌进行化学预防 小鼠模型 (Hi-Myc) 与脂肪酸合成和 β-氧化的抑制有关 从而抑制癌细胞增殖。双盲、随机和安慰剂对照试验 还建议对计划进行前列腺切除术的男性进行机会之窗临床试验，以确定是否 每日口服经过充分表征的 BSE 制剂 (BroccoMax®)，其安全性已得到证实 经临床测试，持续 4 周可抑制循环和前列腺肿瘤的脂肪酸水平。 对上述假设的支持来自我们自己已发表的和初步未发表的研究结果。 具体来说，我们发现 SFN 治疗不仅抑制 ACC1 和 FASN 的蛋白质/mRNA 水平 因为脱氢酶参与脂肪酸的β-氧化，但也会降低人体中乙酰辅酶A的水平 体外前列腺癌细胞和 TRAMP 小鼠体内前列腺肿瘤。乙酰辅酶A是de的构建模块 脂肪酸的新合成，但也在脂肪酸的β-氧化作用下在线粒体中产生。 具体目标：这个高度集中的应用程序的综合具体目标是：（1）确定 使用细胞模型研究 SFN 介导的脂肪酸合成和 β-氧化抑制的机制 前列腺癌和正常前列腺细胞； (2) 确定 SFN 和 SFN 是否能对前列腺癌进行化学预防 Hi-Myc 转基因小鼠中的 BSE 与脂肪酸合成和 β-氧化的抑制有关；和（3） 确定每日口服 BroccoMax® 给药 4 周是否可以减少循环肿瘤和前列腺肿瘤 通过双盲、随机和安慰剂对照机会窗口试点试验来确定脂肪酸水平 计划进行前列腺切除术的男性。 转化影响（意义）：尽管有丰富的机制数据，SFN 或 BSE 的临床发展 像 BroccoMax® 一样，用于前列腺癌的化学预防取决于： (a) 证明 在临床相关的前列腺癌啮齿动物模型中的化学预防功效； (b) 查明 使用前列腺癌的细胞和临床前啮齿动物模型的药效生物标志物； (c) 试点 临床转化研究，以证明对从 细胞/临床前模型，例如本申请中假设的脂肪酸代谢抑制。这 研究的逐步进展对于证明前列腺癌发病率的更大规模临床研究的合理性至关重要 主要终点。该项目的前两个具体目标不仅会测试前列腺癌的可能性 SFN 和 BSE 在与人类分子高度重叠的转基因小鼠模型中进行化学预防 疾病，但还将识别非侵入性药效生物标志物（例如脂肪酸的血清水平） 适用于具体目标 3 中拟议的临床试验。