Vitamin K: Body Pools and Function in Breast Cancer

维生素 K：乳腺癌中的身体储备和功能

• 批准号: 10524195
• 负责人: JoEllen Welsh
• 金额: $ 11.17万
• 依托单位: STATE UNIVERSITY OF NEW YORK AT ALBANY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-02-09 至 2026-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10524195
• 关键词: Adverse effects; Affect; Breast Cancer Cell; Breast Cancer cell line; Cancer Biology; Cell Respiration; Cell model; Cells; Coagulation Process; Data; Data Set; Diet; Dimethylallyltranstransferase; Disease; Energy Metabolism; Enzymes; Gene Expression; Generations; Genes; Growth; Histology; Homeostasis; In Vitro; Intake; Link; Mammary Neoplasms; Mammary gland; Mammospheres; Masks; Measures; Mediating; Metabolism; Mus; Oncogenes; Oxidoreductase; Pathway interactions; Patients; Phenotype; Physiological; Protein Biosynthesis; Proteins; Proteomics; Role; Testing; The Cancer Genome Atlas; Therapeutic; Tissues; Translations; Tumor Biology; Tumor Subtype; Vitamin K; Vitamin K 1; Vitamin K 2; Woman; Xenograft procedure; advanced breast cancer; aggressive breast cancer; aldehyde dehydrogenase 1; bone; cancer genomics; carboxylate; carboxylation; cell growth; clinically relevant; cofactor; dietary; enzyme biosynthesis; feeding; gamma-glutamyl carboxylase; ginsenoside M1; in vivo; insight; loss of function; malignant breast neoplasm; migration; patient subsets; protein function; response; restoration; screening; stem cell biomarkers; stem cells; therapeutic target; triple-negative invasive breast carcinoma; tumor; tumor growth; tumor progression; uptake

PROJECT SUMMARY/ABSTRACT This proposal focuses on the divergent effects of the two major dietary forms of vitamin K on breast cancer. K vitamins act as cofactors for gamma-glutamyl carboxylase (GGCX), which post-translationally introduces γ- carboxyglutamate residues into proteins. Although most of the 17 known γ-carboxylated proteins function in coagulation and bone homeostasis, the presence of GGCX in most tissues (including mammary gland) suggests more extensive physiological roles for vitamin K. We have demonstrated that triple negative breast cancer (TNBC) cell lines express GGCX and produce γ-carboxylated proteins in response to vitamin K1 (phylloquinone), the major dietary form. In TNBC cells, K1 treatment enriches for the stem cell marker aldehyde dehydrogenase 1 (ALDH1) and promotes mammosphere formation. These data suggest that K1 sustains GGXC mediated γ- carboxylation to drive aggressive breast cancer phenotypes. Through analysis of genomic cancer datasets, we find that ~25% of breast tumors express GGCX and the vitamin K oxidoreductase (VKOR) genes required for its activity. Patients with such tumors have poorer survival than those whose tumors do not express these genes at high levels. Patients with this subtype of tumor would be candidates for therapies that limit K1 availability and/or inhibit GGCX. Surprisingly, we found that vitamin K2 (menaquinone-4), another naturally occurring form present in diet, does not stimulate γ-carboxylation or stem cell phenotypes in TNBC cells, but instead strongly suppresses cell growth, migration and energy metabolism. These provocative data indicate that K1 and K2 exert distinct effects on breast cancer cells, with K1 promoting and K2 suppressing aggressive phenotypes. We also found that expression of the vitamin K2 biosynthesis enzyme UbiA Prenyltransferase Domain Containing 1 (UBIAD1) is undetectable in TNBC, suggesting altered cellular handling of vitamin K. In Aim 1 we will dissect the effects of K1 and K2 in vitro, evaluate the role of UBIAD1 and conduct feeding studies to measure accumulation of K1 and K2 in TNBC xenografts and host mammary gland in relation to tumor growth. In Aim 2 we will determine whether deletion of GGCX from TNBC cells impacts γ-carboxylated protein synthesis and aggressive phenotypes in vitro and in vivo. Aim 3 will identify relevant γ-carboxylated GGCX substrate proteins that mediate the effects of K1. We anticipate that growth of tumors with high GGCX activity and low UBIAD1 will be stimulated by high dietary K1 and inhibited by high dietary K2. These findings would identify GGCX as an oncogene and the vitamin K pathway as a therapeutic target in a subset of patients with advanced breast cancer.

项目概要/摘要 该提案的重点是维生素 K 的两种主要膳食形式对乳腺癌的不同影响。 K 维生素充当 γ-谷氨酰羧化酶 (GGCX) 的辅助因子，该酶在翻译后引入 γ- 羧基谷氨酸残基转化为蛋白质。尽管 17 种已知的 γ-羧化蛋白中的大多数在 凝血和骨稳态，大多数组织（包括乳腺）中存在 GGCX 表明 维生素 K 具有更广泛的生理作用。我们已经证明，三阴性乳腺癌 (TNBC) 细胞系表达 GGCX 并产生 γ-羧化蛋白以响应维生素 K1（叶绿醌）， 主要的饮食形式。在 TNBC 细胞中，K1 处理可富集干细胞标记物醛脱氢酶 1 (ALDH1) 并促进乳腺球形成。这些数据表明 K1 维持 GGXC 介导的 γ- 羧化驱动侵袭性乳腺癌表型。通过对癌症基因组数据集的分析，我们 发现约 25% 的乳腺肿瘤表达 GGCX 和其所需的维生素 K 氧化还原酶 (VKOR) 基因 活动。患有此类肿瘤的患者比那些肿瘤不表达这些基因的患者的生存率更差 高水平。患有这种肿瘤亚型的患者将是限制 K1 可用性和/或 抑制 GGCX。令人惊讶的是，我们发现维生素 K2（menaquinone-4）是另一种天然存在的形式 在饮食中，不会刺激 TNBC 细胞中的 γ-羧化或干细胞表型，而是强烈抑制 细胞生长、迁移和能量代谢。这些令人兴奋的数据表明 K1 和 K2 发挥着不同的作用 对乳腺癌细胞的影响，K1 促进侵袭性表型，K2 抑制侵袭性表型。我们还发现 维生素 K2 生物合成酶 UbiA Prenyltransferase Domain Containing 1 (UBIAD1) 的表达 在 TNBC 中检测不到，表明细胞对维生素 K 的处理发生了改变。在目标 1 中，我们将剖析其影响 K1 和 K2 体外分析，评估 UBIAD1 的作用并进行喂养研究以测量 K1 的积累 TNBC 异种移植物和宿主乳腺中的 K2 和 K2 与肿瘤生长的关系。在目标 2 中，我们将确定 从 TNBC 细胞中删除 GGCX 是否会影响 γ-羧化蛋白合成和侵袭性 体外和体内的表型。目标 3 将鉴定介导相关的 γ-羧化 GGCX 底物蛋白 K1的影响。我们预计，具有高 GGCX 活性和低 UBIAD1 的肿瘤生长将会受到刺激 受高膳食 K1 的影响，并受高膳食 K2 的抑制。这些发现将确定 GGCX 是一种癌基因，并且 维生素 K 通路作为晚期乳腺癌患者子集的治疗靶点。