Novel Markers for Disease Outcome in Breast Cancer

乳腺癌疾病结果的新标志物

• 批准号: 8552879
• 负责人: Stefan Ambs
• 金额: $ 41.8万
• 依托单位: DIVISION OF BASIC SCIENCES - NCI
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8552879
• 关键词: Adrenergic Agents; Affect; Africa; African; African American; American; Asians; Award; Biochemical; Biological; Biological Assay; Biological Markers; Blood specimen; Body mass index; Breast Cancer Cell; Breast Diseases; CD44 Antigens; CD44 gene; Cancer Patient; Cancer Research Project; Cancer cell line; Cell Culture System; Cells; Characteristics; Chicago; Clinical; Clinical Research; Collaborations; Consent; DNA Modification Process; Data; Development; Diagnostic Neoplasm Staging; Disease; Disease Marker; Disease Outcome; Disease Progression; Disease susceptibility; ERBB2 gene; Epidermal Growth Factor Receptor; Estrogen receptor negative; European; Event; Freezing; Frequencies; Funding; Gene Expression; Gene Expression Profile; Gene Expression Profiling; Genetic Predisposition to Disease; Genetic Transcription; Genetic Variation; Goals; IL8 gene; Income; Laboratories; Life; Link; Lipids; Malignant Neoplasms; Malignant neoplasm of lung; Mammary Gland Parenchyma; Mammary Neoplasms; Medicine; Menopausal Status; Modeling; Molecular; Mutation; NOS2A gene; Names; Nitric Oxide; Nitric Oxide Signaling Pathway; Normal tissue morphology; Outcome; Oxidation-Reduction; Pathway interactions; Patient Education; Patients; Pattern; Phenotype; Pilot Projects; Population Group; Predisposition; Production; Property; Proteins; Proteome; Publishing; Radiation therapy; Reporting; Research; Research Personnel; Resistance; Signal Transduction; Stem cells; Stress; Stressful Event; Surveys; Testing; Tissue Sample; Tissues; Tumor Biology; Tumor Markers; Tumor Subtype; Tumor stage; Universities; Up-Regulation; Vascularization; Winking; Woman; Work; Wound Healing; adrenergic; angiogenesis; c-myc Genes; cancer surgery; carcinogenesis; clinically significant; college; cyclooxygenase 2; design; embryonic stem cell; experience; follow-up; genome wide association study; health disparity; human NOS2A protein; inflammatory breast cancer; inhibitor/antagonist; innovation; malignant breast neoplasm; mortality; new therapeutic target; novel; novel marker; osteopontin; outcome forecast; overexpression; prognostic; response; stem cell division; triple-negative invasive breast carcinoma; tumor; validation studies

Inducible nitric oxide synthase (NOS2) and cyclooxygenase-2 are involved in wound healing, angiogenesis, and carcinogenesis. NOS2 up-regulation and increased nitric oxide (NO) production also affects the redox state of cells and induces protein, lipid, and DNA modifications. Recent research by our laboratory led to the novel and clinically significant observation that NOS2 expression is associated with a prognostic basal-like transcription pattern and is an independent predictor of poor survival in women with ER-negative breast tumors. NOS2 remained a predictor of poor survival when the analysis was restricted to only those patients with basal-like breast tumors. This result suggests that in addition to inducing a basal-like signature in ER-negative tumors, NOS2 may further enhance disease aggressiveness in the presence of this signature. Increased NOS2 in breast tumors also correlated with other poor outcome markers, such as an increased tumor vascularization and p53 mutation frequency, and activated EGFR. Both p53 mutations and EGFR overexpression occur more commonly in basal-like breast tumors than other breast cancer subtypes. Additional work showed that nitric oxide activates EGFR, consistent with previous findings in lung cancer, and induces proteins, such as CD44 and c-Myc, in ER-negative human breast cancer cells. These markers have been linked to an embryonic stem cell-like phenotype in breast cancer and disease outcome. Recently, an increased expression of stem cell markers in basal-like tumors has been reported. Among them was CD44, which is a receptor for hyaluronan and osteopontin. CD44 is a poor outcome marker in breast cancer and CD44-positive breast cancer cells have an increased invasive activity, increased resistance to radiation therapy and chemotherapeutics. Together, these novel observations link NOS2 to the development of a poorly differentiated breast cancer phenotype with stem cell-like characteristics. NO may induce this phenotype by activation of c-Myc or by inducing the release of stem cell renewal factors like IL-8, a property that nitric oxide has, as we have shown. We propose that NOS2 and downstream targets of NOS2 signaling are novel therapeutic targets for ER-negative breast cancer in general, and more specifically for basal-like breast cancer and the triple-negative disease. These findings are currently further pursued in collaboration with the laboratory of Dr. David Wink at the NCI. His group is studying the NO signaling pathways in ER-negative breast cancer cell lines and how these pathways induce a basal-like and stem cell-like phenotype in these cells. In addition, our clinical findings are possibly followed up as part of a Major Opportunity initiative at the NCI in which the Clinical Center (PI: Elise Kohn) will test a NOS2-specific inhibitor provided by GlaxoSmithKline in patients with triple-negative and inflammatory breast cancer. In FY12, we continued to comprehensively examine the metabolome, proteome and transcriptome of ER-positive and ER-negative breast tumors from African-American and European-American patients for biomarker discovery. This project received funds through a NCI Director Innovation Award. The promise of the study is the discovery of novel biomarkers for prognosis, and for elucidating what may drive the aggressiveness of breast cancer in African-American women. To date, we have completed the metabolome and transcriptome analysis of the tumors and the adjacent non-cancerous tissues and have collected the raw data from the proteome analysis for about 75% of the tissues. The transcriptome data are being analyzed and compared with data from a previously published pilot study (Martin et al. PLoS One, 2009), showing significant gene expression differences between tumors from African-American and European-American breast cancer patients. The analysis of the metabolome yielded preliminary information on 536 biochemicals in the breast tumors and the surrounding normal tissues. Of those, 333 are named/identified. 360 biochemicals differed significantly in their concentrations between ER-negative and normal surrounding tissue. 311 differed in their concentrations between ER-positive and normal surrounding tissue. There were significant differences in metabolite levels between breast cancer subtypes and tumors from African-American patients and European-American patients, specifically in ER-negative tumors. We did not find any differences in tissue metabolite concentrations by body mass index, menopausal status, or by tumor stage and tumor HER2 status. There are perhaps also differences by income and education of the patients, a finding which will need further examination. Currently, we are validating these findings in a follow-up study consisting of 106 estrogen receptor-negative tumors and adjacent non-cancerous tissues (as available) from African-American (n = 49) and European-American breast cancer patients (n = 21). In this study, we will evaluate the abundance of those 80-150 metabolites that showed the most significant differences in the pilot study between tumor and normal and between African-American and European-American patients. This validation study is conducted at the Baylor College of Medicine in collaboration with Arun Sreekumar, using targeted assays. In addition, we are examining the influence of candidate metabolites on cancer phenotypes in cell culture systems.In a collaboration with Dr. Olopade at the University of Chicago and investigators from other academic centers, we are participating in a GWAS study of breast cancer in women of African ancestry. This study is aimed to identify genetic susceptibility loci in African-American women and women from West Africa. First results show that many of the disease susceptibility loci first discovered for European and European-American women cannot be replicated in women of African descent. Future research will focus on novel susceptibility loci that have not been detected in other population groups (of European or Asian descent).Lastly, we started a new project evaluating the impact of stressful life events on tumor biology. In cell culture systems, we will model the effects of beta adrenergic signaling on tumor stroma interactions. In a clinical study, we will give breast cancer patients, who have breast cancer surgery, a short survey evaluating their perceived stress within the last one month. We will also collect frozen tumor and adjacent normal breast tissue and blood samples from these patients and evaluate whether the breast tissue or the blood samples have a biological signatures related to their perceived stress. We hypothesize that patients with a high perceived stress exposure have a biological signature consistent with a more aggressive disease and poorer survival. The pilot study is designed to collect 100 tumor/normal pairs from consented patients with a completed survey.

诱导型一氧化氮合酶（NOS2）和环氧酶-2参与伤口愈合，血管生成和致癌作用。 NOS2上调和一氧化氮（NO）产生增加还会影响细胞的氧化还原状态，并诱导蛋白质，脂质和DNA修饰。我们实验室的最新研究导致了新颖且临床上显着的观察结果，即NOS2表达与预后的基础样转录模式有关，并且是ER阴性乳腺肿瘤女性生存不良的独立预测指标。当分析仅限于那些基底样乳腺肿瘤的患者时，NOS2仍然是存活不良的预测指标。该结果表明，除了在ER阴性肿瘤中诱导基底样特征外，NOS2还可以在这种特征的存在下进一步提高疾病的侵略性。乳腺肿瘤中NOS2的增加也与其他较差的预后标志物相关，例如肿瘤血管增加和p53突变频率和激活的EGFR。与其他乳腺癌亚型相比，p53突变和EGFR过表达在基础样乳腺肿瘤中更常见。其他工作表明，一氧化氮激活EGFR，与先前的肺癌发现一致，并在ER阴性的人类乳腺癌细胞中诱导蛋白质（例如CD44和C-MYC）。这些标记与乳腺癌和疾病预后中的胚胎干细胞样表型有关。最近，据报道，干细胞标记物在基底样肿瘤中的表达增加。其中包括CD44，它是透明质酸和骨桥蛋白的受体。 CD44在乳腺癌中的预后标志很差，CD44阳性乳腺癌细胞具有增加的侵入性活性，对放射治疗和化学治疗的耐药性增加。这些新颖的观察结果将NOS2与干细胞样特征的开发联系起来。否可以通过激活C-MYC或诱导干细胞更新因子（例如IL-8）释放这种表型，如我们所示，这是一氧化氮具有的特性。我们建议NOS2信号的NOS2和下游靶标通常是ER阴性乳腺癌的新型治疗靶标，更专门针对基底样乳腺癌和三阴性疾病。 目前，这些发现与NCI的David Wink博士实验室合作进一步追求。他的小组正在研究ER阴性乳腺癌细胞系中的NO信号通路，以及这些途径如何在这些细胞中诱导基础样和干细胞样表型。此外，我们的临床发现可能是在NCI的一项主要机会计划的一部分中进行的，其中临床中心（PI：Elise Kohn）将测试由GlaxoSmithkline在三个阴性和炎性乳腺癌患者中提供的NOS2特异性抑制剂。 在2012财年，我们继续全面研究来自非裔美国人和欧洲裔美国人生物标志物发现的ER阳性和ER阴性乳腺肿瘤的代谢组和转录组。该项目通过NCI主管创新奖获得了资金。该研究的希望是发现了预后的新型生物标志物，并阐明了可能推动非裔美国妇女乳腺癌侵略性的原因。 迄今为止，我们已经完成了肿瘤和相邻非癌组织的代谢组和转录组分析，并从蛋白质组分析中收集了约75％的组织中的原始数据。转录组数据正在分析，并将其与先前发表的初步研究（Martin等人PLOS One，2009）的数据进行了比较，显示了来自非裔美国人和欧美乳腺癌患者肿瘤之间的显着基因表达差异。对代谢组的分析产生了有关乳腺肿瘤和周围正常组织中536种生化物质的初步信息。其中333个被命名/识别。 360生物化学物质在ER阴性和正常组织之间的浓度上显着差异。 311在ER阳性和正常周围组织之间的浓度有所不同。乳腺癌亚型和非裔美国人患者和欧洲裔美国人，特别是在ER阴性肿瘤中的乳腺癌亚型和肿瘤之间存在显着差异。我们没有发现组织代谢物浓度的差异，体重指数，更年期状态或肿瘤期和肿瘤HER2状态。可能还会因患者的收入和教育而有所不同，这一发现需要进一步检查。 目前，我们正在一项随访研究中验证这些发现，该研究由106个雌激素受体阴性肿瘤和非裔美国人（n = 49）和欧美乳腺癌患者（n = 21）组成的雌激素受体阴性肿瘤和邻近的非癌组织（可用）。在这项研究中，我们将评估80-150代谢产物的丰度，这些代谢物在肿瘤与正常情况下以及非裔美国人和欧美患者之间的试验研究中最显着差异。这项验证研究是在贝勒医学院与Arun Sreekumar合作的。此外，我们正在研究候选代谢物对细胞培养系统中癌症表型的影响。在与芝加哥大学的Olopade博士以及其他学术中心的研究人员的合作中，我们正在参加非洲血统女性的GWAS研究。这项研究旨在确定来自西非的非裔美国妇女和妇女的遗传易感性基因座。第一个结果表明，在非洲血统的妇女中，无法复制许多针对欧洲和欧美妇女发现的疾病敏感性基因座。 未来的研究将重点放在其他人口群体（欧洲或亚洲血统）中尚未检测到的新型易感基因座。最终，我们开始了一个新的项目，评估了压力性生命事件对肿瘤生物学的影响。在细胞培养系统中，我们将模拟β肾上腺素能信号传导对肿瘤基质相互作用的影响。 在一项临床研究中，我们将给予接受乳腺癌手术的乳腺癌患者，这是一项简短的调查，评估了在过去一个月内的感知压力。我们还将从这些患者那里收集冷冻的肿瘤和邻近的正常乳腺组织以及血液样本，并评估乳腺组织或血液样本是否具有与其感知胁迫有关的生物学特征。我们假设，感知到高的压力暴露的患者具有与更具侵略性疾病和较差生存率一致的生物学特征。该试点研究旨在从同意的患者中收集100对肿瘤/正常对，并完成了完整的调查。