Vitamin D and Progestins for the Chemoprevention of Fallopian Tube/Ovarian Cancer

维生素 D 和孕激素用于输卵管/卵巢癌的化学预防

• 批准号: 10302294
• 负责人: Gustavo Rodriguez
• 金额: $ 35万
• 依托单位: NORTHSHORE UNIVERSITY HEALTHSYSTEM
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-12-01 至 2024-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10302294
• 关键词: Abdominal Cavity; Aftercare; Animal Model; Apoptosis; Apoptotic; Binding; Biological Markers; CA-125 Antigen; CRISPR/Cas technology; Calcitriol; Cancer cell line; Carcinoma; Carcinoma in Situ; Catabolism; Cell Death; Cell Line; Cells; Cessation of life; Chemoprevention; Chemopreventive Agent; Cholecalciferol; Data; Development; Disabled Persons; Electrophoretic Mobility Shift Assay; Epithelial; Epithelial Cells; Epithelial ovarian cancer; Exhibits; Female; Genes; Glycoproteins; Goals; Gynecologic; Half-Life; Hormone use; Human; In Vitro; Incidence; Invaded; Large T Antigen; Lead; Lesion; Life; Light; Luciferases; Malignant Female Reproductive System Neoplasm; Malignant Neoplasms; Malignant neoplasm of fallopian tube; Malignant neoplasm of ovary; Mammalian Oviducts; Mediating; Mesenchymal; Messenger RNA; Metabolism; Mifepristone; Modeling; Molecular; Molecular Target; Mouse Strains; Mus; Mutation; Neoplasm Metastasis; Oligonucleotides; Oral Contraceptives; Ovarian; Ovary; PTEN gene; Pathway interactions; Pharmacology; Phenotype; Prevention; Prevention approach; Preventive; Process; Production; Progesterone; Progesterone Receptors; Progestins; Proteins; Publishing; Reporter; Research; Response Elements; Risk; Serous; Simian virus 40; Site; Structure of paramesonephric duct; Surface; Surrogate Endpoint; TGF Beta Signaling Pathway; TP53 gene; Testing; Tissues; Transforming Growth Factor beta; Tube; Vitamin D; Vitamin D3 Receptor; Woman; Xenograft procedure; base; cancer chemoprevention; cancer prevention; carcinogenesis; cell immortalization; cell injury; design; efficacy testing; experimental study; genome editing; in vivo; inhibitor; mouse model; novel; ovarian cancer prevention; pre-clinical; prevent; progesterone receptor negative; progesterone receptor positive; promoter; response; stable cell line; synergism; tumor; tumorigenesis

Project Summary/Abstract There is significant potential to decrease ovarian cancer incidence through prevention. Oral contraceptives (OCs) confer a 30-50% reduced risk of developing epithelial ovarian cancer, suggesting that an effective preventive approach using hormones is possible. Progestin-potent OCs confer twice the ovarian cancer protection as newer weak-progestin OCs. These data suggest that a progestin-based pharmacologic strategy may be highly effective for prevention of ovarian cancer. Progestin potency can be enhanced through combination with vitamin D, a second preventive agent that is also non-toxic when administered as cholecalciferol; therefore, the two agents may act synergistically on chemopreventive endpoints in the gynecologic tract such that progestins positively influence vitamin D metabolism. The tissue of origin of ovarian cancer remains uncertain. There is now compelling evidence that the origin of many high grade ovarian cancers may be the fallopian tube, thus creating an opportunity for identifying molecular targets in the fallopian tube for effective ovarian cancer prevention. The proposed research is designed to determine the molecular mechanisms by which ovarian cancer, whether it originates from the ovary or the fallopian tube, may be prevented. Prevention of fallopian tube/ovarian cancer using a combination of vitamin D and each of the three classes of progestins will be tested in the following three aims. First, the impact of these treatments on molecular pathways that lead to cell death in fallopian tube and ovarian epithelial cells will be investigated. This will include extension of the active life of vitamin D by inhibition of CYP24A1, as well as increased sensitivity of these cells to vitamin D by enhanced production of the vitamin D receptor. Secondly, the interaction between the progesterone receptor and progesterone response elements PRE(s) will be mechanistically defined in the CYP24A1 promoter through the use of CYP24A1 promoter-luciferase reporter constructs. Endogenous functional PRE(s) in ovarian cancer cell lines will be selectively inactivated using CRISPR/Cas9-mediated genome editing. The resulting stable cell line(s) harboring non-functional PRE(s) in the CYP24A1 promoter will then be used to validate loss of synergy between progesterone and vitamin D in vitro as well as in vivo in mouse xenografts. Thirdly, in vivo experiments using two genetically modified mouse strains that develop fallopian tube cancer closely resembling human ovarian cancer will be performed to determine the in vivo impact of progestins and vitamin D on fallopian tube epithelial cancer development. Understanding the mechanisms by which progestins are integrally involved in each of these processes, will facilitate development of a robust pharmacologic approach to prevent ovarian/fallopian tube cancer. The goal of this research is to identify an optimal pharmacologic strategy combining progestins and vitamin D for an effective ovarian cancer chemopreventive approach that can be tested in women at risk of developing ovarian cancer.

项目摘要/摘要 通过预防降低卵巢癌发病率的潜力很大。口服避孕药 （OCS）赋予30-50％的降低患卵巢癌的风险，这表明有效 可以使用激素的预防方法。孕激素功能OC赋予卵巢癌的两倍 作为较新的弱蛋白OCS保护。这些数据表明基于孕激素的药物策略 对于预防卵巢癌可能是非常有效的。孕激素效力可以通过 与维生素D结合使用，这是第二种预防剂，当给药时也无毒 胆固醇；因此，两种代理可以在化学预防终点协同作用 妇科学使孕激素阳性地影响维生素D代谢。卵巢的起源组织 癌症仍然不确定。现在有令人信服的证据表明许多高级卵巢的起源 癌症可能是输卵管，因此为识别输卵管中的分子靶标创造了机会 用于预防有效卵巢癌的管。拟议的研究旨在确定分子 卵巢癌（无论是起源于卵巢还是输卵管）的机制可能是 阻止。使用维生素D和三种组合预防输卵管/卵巢癌 孕激素类别将在以下三个目标中进行测试。首先，这些治疗对 将研究导致输卵管和卵巢上皮细胞中细胞死亡的分子途径。 这将包括通过抑制CYP24A1的延长维生素D的活性寿命，以及增加 通过增强维生素D受体的产生，这些细胞对维生素D的敏感性。其次， 孕酮受体与孕酮反应元件之间的相互作用是 通过使用CYP24A1启动子 - 卢西酶报道器在CYP24A1启动子中定义的机械定义 构造。卵巢癌细胞系中的内源性功能性PRE将被选择性地灭活 CRISPR/CAS9介导的基因组编辑。由此产生的稳定细胞系（s）在 然后，CYP24A1启动子将用于验证孕酮和维生素D之间的协同作用损失 小鼠异种移植物中的体外和体内。第三，使用两种基因修饰小鼠的体内实验 患有输卵管癌的菌株将与人类卵巢癌紧密相似 确定孕激素和维生素D对输卵管上皮癌发育的体内影响。 了解孕激素与每个过程中的每个过程中的每个过程中的机制，将 促进开发可预防卵巢/输卵管癌的强大药理方法。目标 这项研究的是确定一种结合孕激素和维生素D的最佳药理策略 有效的卵巢癌化学预防方法可以在患有卵巢的风险的女性中进行测试 癌症。