Identification of metabolic adducts associated with prostate cancer progression in African American men

摘要 前列腺癌（PCa）是男性癌症相关死亡的第二大原因。非裔美国人/黑人 (AA/B）男性受前列腺癌的影响，发病率高60%， 与欧洲裔美国人（EA）男性相比，死亡风险增加。迫切需要查明 潜在的生物学变化，导致这种差异，以开发包容性的诊断和预测测试 和量身定制的治疗方法。驱动PCa健康的生物学变化的原因很多 已经提出了差异，包括社会经济、遗传和环境因素。这些因素都 引起代谢改变，这是一种与PCa发作和进展相关的生物学变化。一项拟议 这些变化驱动PCa的机制是通过产生反应性亲电试剂丙酮醛 （MG）. MG是脂质、蛋白质和糖代谢的副产物，并在DNA、RNA和糖上形成共价加合物。 蛋白这些加合物，称为MG-晚期糖基化终产物（MG-AGEs），导致DNA突变， 基因组的不稳定性，改变RNA的稳定性和翻译，并改变蛋白质的稳定性和功能。此外，MG- AGEs结合并激活AGEs受体（AGEs）。为了调节MG和MG-AGEs，细胞使用谷胱甘肽酶1 （GLO 1）以解毒MG和可溶性AGEs（sNAG）以螯合MG-AGEs并防止AGEs活化。这些 这些分量被称为AGE/Age轴。我们的长期目标是确定千年发展目标-老龄化群体的作用， 年龄/年龄轴作为PCa的生物标志物和驱动因素，并确定种族差异如何影响这一过程。 为了明确MG-AGEs、GLO 1、β-淀粉样蛋白和sIgA与前列腺癌健康差异的关系，我们设计了一个 AA/B和EA男性伴和不伴PCa的巢式病例对照试验。我们用质谱法测定血清MG-AGEs， 用荧光分光光度法、ELISA法测定血清学，并对100名受试者的GLO 1和AGER（编码AGER的基因）基因座进行测序。 从全血中分离的基因组DNA。我们发现MG-AGEs、sNAG、GLO 1和AGER SNPs 在AA/B型男性中与PCa显著相关，但在EA型男性中与PCa无关。我们还观察到一个显著的差异， AA/B组与EA组无PCa组比较，差异有统计学意义（P < 0. 05）。这让我们假设MG-AGEs， sIgA、GLO 1和AGER SNPs可用作AA/B男性中PCa生物标志物，GLO 1 SNPs 可能在MG-AGEs的积累、突变和PCa细胞生长中起作用。为了验证这个假设，我们 建议1）使用AGE/ESTA轴的分子和遗传特征沿着人口统计学和临床 使用多变量临床-遗传风险模型预测AA/B和EA男性PCa风险的变量，以及2） 使用来自AA/B和EA PCa肿瘤的细胞系来确定GLO 1 SNP对MG-AGE的影响 在某些实施方案中，所述方法包括抑制肿瘤细胞的增殖、蓄积、细胞生长、转移标志物的表达和基因组突变的诱导。这 本工作首次采用PCA法对AA/B和EA患者的MG-AGEs进行了分析，采用了新的质谱法 方法，并描述了来自不同种族群体的表达GLO 1多态性的细胞系的产生。

Abstract Prostate cancer (PCa) is the second highest cause of cancer-related deaths in men. African American/Black (AA/B) men are disproportionally impacted by PCa with a 60% higher incidence of disease and a 2-3x fold increase in mortality risk compared to European American (EA) men. There is an urgent need to identify the underlying biological changes that give rise to this disparity to develop inclusive diagnostic and predictive tests and tailored therapeutic treatments. A myriad of causes for the biological changes that drive PCa health disparities have been proposed, including socioeconomic, genetic, and environmental factors. These factors all give rise to altered metabolism, a biological change associated with PCa onset and progression. A proposed mechanism for how these changes drive PCa is through the production of the reactive electrophile methylglyoxal (MG). MG is a by-product of lipid, protein, and sugar metabolism and forms covalent adducts on DNA, RNA, and protein. These adducts, termed MG-advanced glycation end products (MG-AGEs) lead to DNA mutations and genomic instability, change RNA stability and translation, and alter protein stability and function. In addition, MG- AGEs bind and activate the receptor for AGEs (RAGE). To regulate MG and MG-AGEs, cells use glyoxalase 1 (GLO1) to detoxify MG and soluble RAGE (sRAGE) to sequester MG-AGEs and prevent RAGE activation. These components are termed the AGE/RAGE axis. Our long-term goal is to define the role of MG-AGEs and the AGE/RAGE axis as biomarkers and drivers of PCa and determine how racial disparities influence this process. To define the association of MG-AGEs, GLO1, RAGE, and sRAGE with PCa health disparities, we designed a nested case-control trial of AA/B and EA men with and without PCa. We measured serum MG-AGEs using mass spectrometry, serum sRAGE using ELISA, and sequenced the GLO1 and AGER (gene encoding RAGE) loci in genomic DNA isolated from whole blood. We discovered that MG-AGEs, sRAGE, and GLO1 and AGER SNPs were significantly associated with PCa in AA/B men but not EA men. We also observed a significant difference between these components in AA/B and EA men without PCa. This led us to hypothesize that MG-AGEs, sRAGE, and GLO1 and AGER SNPs may have utility as biomarkers for PCa in AA/B men and that GLO1 SNPs may play a role in the accumulation of MG-AGEs, mutations, and PCa cell growth. To test this hypothesis, we propose to 1) use molecular and genetic features of the AGE/RAGE axis along with demographic and clinical variables to predict the risk of PCa in AA/B and EA men using a multivariable clinical-genetic risk model and 2) use cell lines derived from AA/B and EA PCa tumors to determine the impact of GLO1 SNPs on MG-AGE accumulation, cell growth, the expression of metastatic markers, and the induction of genomic mutations. This work represents the first analysis of MG-AGEs in AA/B and EA men with PCA, utilizes novel mass spectrometry methods, and describes generation of cell lines expressing GLO1 polymorphisms from diverse racial groups.