Biomarkers of Human Lung Cancer

人类肺癌的生物标志物

• 批准号: 8349212
• 负责人: Curtis Harris
• 金额: $ 60.13万
• 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/8349212
• 关键词: 19p; 19p13.3; Accounting; Adenocarcinoma; Adjuvant; Adjuvant Chemotherapy; Alleles; Anabolism; Biological Markers; Cancer cell line; Cancerous; Cells; Chromosomes; Clinical; Clinical Trials; Code; Coupled; DNA; DNA copy number; Data; Development; Diagnosis; Diagnostic; Diagnostic Neoplasm Staging; Disease; Early Diagnosis; Estrogen Receptor alpha; Estrogen Receptors; Estrogens; Family; Female; Formalin; Freezing; Gender; Genes; Genetic; Genetic Polymorphism; Genetic Transcription; Germ Lines; Goals; Histocompatibility Testing; Hormone Receptor; Hormone replacement therapy; Human; In Vitro; International; Japan; Japanese Population; Lead; Loss of Heterozygosity; Lung Adenocarcinoma; Lung Neoplasms; Malignant Neoplasms; Malignant Pleural Mesothelioma; Malignant neoplasm of lung; Maps; Maryland; Measures; Messenger RNA; MicroRNAs; Molecular; Mutation; Non-Small-Cell Lung Carcinoma; Normal tissue morphology; Norway; Oncogenic; Outcome; Paraffin Embedding; Pathway interactions; Patients; Plasma; Pleural Mesothelial Cell; Pleural Mesothelioma; Predictive Value; Progesterone; Protein-Serine-Threonine Kinases; RNA; Randomized; Relapse; Reporting; Resected; Role; STK11 gene; Sampling; Serum; Simian virus 40; Single Nucleotide Polymorphism; Smoker; Somatic Mutation; Specimen; Staging; Systems Biology; TNM; Telomerase; Telomerase Inhibitor; Telomere Length Maintenance; Testing; Tissues; Translations; Tumor Suppressor Genes; Tumor stage; Variant; base; burden of illness; cancer initiation; cancer therapy; carcinogenesis; chromosome loss; cohort; improved; in vitro Model; in vivo; male; mortality; novel; outcome forecast; prognostic; telomere; therapeutic target; tumor; tumor progression; tumorigenesis

Early detection of primary lung cancer is difficult yet important since diagnosis at earlier stages is associated with significantly better survival. Circulating microRNA (miR) profiles have been proposed as promising diagnostic and prognostic biomarkers for cancer, including lung cancer. The goal in this study was to establish biomarkers for early detection of primary lung cancer (NSCLC) by measuring circulating miRs. We looked at paired serum and plasma samples of patients with early stage NSCLC and matched controls. We discovered differential expression of the following miRs: miR-146b, miR-221, let-7a, miR-155, miR-17-5p, miR-27a, and miR-106a, which were significantly reduced in the serum of NSCLC cases, while miR-29c was significantly increased. Even though we did not observe any significant differences in plasma, and expression levels in serum did not correlate well with levels of plasma, these tissue types are obtained differently, which could account for differences in the results. Moreover, reduced plasma expression of let-7b was modestly associated with worse cancer-specific mortality in all patients, whereas reduced serum expression of miR-223 was modestly associated with cancer-specific mortality in stage IA/B patients. Additionally, we looked at the expression of biologically relevant miRs (miR-21, miR-29b, miR-34a/b/c, miR-155, and let-7a) in formalin-fixed paraffin-embedded tumor specimens from the patients who participated in the International Adjuvant Lung Cancer Trial (IALT), the largest randomized study conducted to date of adjuvant chemotherapy in patients with radically resected NSCLC. The goal was to evaluate if the expression of these miRs represents prognostic and predictive value for survival. Importantly, the expression of certain miRs has been associated with chemoresistance. However, our results indicate that the miRs expression patters examined were neither predictive nor prognostic in this large patient cohort. Based on the previous miR microarray data of lung cancer specimens from NSCLC adenocarcinoma patients, we investigated the expression of 5 miRs whose expression were significantly altered in lung cancer and were associated with cancer-specific mortality. Three of those miRs (miR-17, miR-21, and miR-155) were potential oncogenic miRs, showing increased expression in tumors with high expression levels being associated with poor prognosis. We investigated their expression in early-stage lung adenocarcinoma snap-frozen tissues from NSCLC patients that originated from Maryland, Norway and Japan in association with tumor progression and survival. We found elevated expression of all three miRs to be associated with worse cancer-specific mortality in the Maryland cohort. However, only miR-21 showed the same association in the Norwegian cohort and worse relapse-free survival in the Japanese cohort. Additionally, more advanced stage tumors expressed significantly higher levels of miR-21 compared with TNM stage I patients, in which group miR-21 was also associated with worse cancer-specific mortality and relapse-free survival, independent of other clinical factors. We found that elevated miR-21 expression is independently associated with worse prognosis in 3 independent NSCLC adenocarcinoma cohorts from different regions of the world. This finding has a translational relevance; miR-21 expression has potential utility as an early-stage prognostic biomarker for NSCLC adenocarcinoma patients. Although we did not observe a significant increase of miR-21 expression in FFPE tissues, this could be due to lower quality of RNA extracted from those tissues, as has been reported previously. Moreover, we have undertaken a systems biology approach to study the effect of miRs in cancer. Each miR can control translation of hundreds of different coding messengers, and a single messenger can be controlled by more than one miR. We combined differential expression, genetic networks and DNA copy number alterations. We confirmed, or discovered, miRs with comprehensive roles in cancer: has-miR-103/106 were downregulated in cancer, whereas has-miR-30 became most prominent; has-miR-17/92 family was amplified and the has-miR-143/145 cluster deleted in cancer; has-miR-30 and has-miR-204 were found to be physically altered at the DNA copy number level. We reported the first miR network from normal tissues, but also built miR networks for coupled cancerous and noncancerous tissues, and identified cancer variations in miR networks. Finally, we superimposed DNA variations onto expression data to generate a comprehensive miR alteration map in cancer. In addition to our studies with miRs as important biomarkers in lung cancer, we investigated a tumor suppressor gene, LKB1/STK11, which has a serine-threonine kinase activity and is located on chromosome 19p13.3. Chromosome 19p is the second most common region of loss in lung tumors of smokers, and LKB1 mutations are found in 30% of lung cancer cell lines and in a smaller portion of primary lung adenocarcinomas. In this study, we used several complementary genetic approaches to assess the KLB1 locus in primary NSCLC. Our data identified total inactivation of the LKB1 gene by either homozygous deletion (HD) or loss of heterozygosity (LOH) with somatic mutation in 39% of tested samples, whereas loss of chromosome 19p region by HD or LOH at the LKB1 region occurred in 90% of NSCLC. Furthermore, we investigated the role of tumor estrogen receptors (ERs) and serum estrogen in lung cancer with a hypothesis that ERs and functional single nucleotide polymorphisms (SNPs) in the estrogen biosynthesis pathway are associated with poorer lung cancer survival. Serum estrogen, progesterone, tumor messenger RNA expression of hormone receptors and germ line DNA polymorphisms were analyzed for associations with lung cancer survival. Patients in the highest tertile of serum estrogen had worse survival in all three cohorts investigated in this study. Furthermore, the variant allele of estrogen receptor alpha (ER-alpha) polymorphism (rs2228480) was significantly associated with increased tumor ER-alpha levels and worse survival in all cohorts. Results were independent of gender and hormone replacement therapy. Thus, we reported a significant association of increased serum estrogen with poorer survival among lung cancer male and female patients. Additionally, we investigated a very important mechanism, cellular immortalization, which is one of the hallmarks of cancer and is dependent on the activity of a telomere length maintenance mechanism (TMM), either telomerase or alternative lengthening of telomeres (ALT). We examined 43 malignant pleural mesotheliomas (MPMs). The TMMs are widely regarded as potential targets for cancer therapies and telomerase inhibitors have entered clinical trials. We asked what proportion of MPMs use ALT and/or telomerase. We found that 43 of 43 MPMs were telomerase-positive[+] and Alt-negative[-]. To investigate whether MPM cells are unusually susceptible to activation of telomerase, we examined activation of the TMMs in an in vitro model of cellular immortalization, in which normal pleural mesothelial cells were transduced with SV40. We discovered that pleural mesothelioma cells are capable of activating either TMM in vitro, and the observation that 100% MPMs were telomerase[+] suggests that there are factors in vivo that select for telomerase activity during oncogenesis of this tumor type. We concluded that MPM is a tumor that could be considered for anti-telomerase therapy. Genes and mechanisms described in our studies, important to lung cancer initiation, progression and outcome, present potential molecular prognostic and diagnostic biomarkers in lung cancer.

早期检测原发性肺癌很困难，但由于早期阶段的诊断与生存率明显更好。已经提出循环microRNA（miR）特征是包括肺癌在内的癌症的有希望的诊断和预后生物标志物。这项研究的目的是通过测量循环miR来建立生物标志物，以早日检测原发性肺癌（NSCLC）。我们研究了早期NSCLC和匹配对照的患者的配对血清和血浆样品。我们发现了以下miR的差异表达：miR-146b，miR-221，let-7a，miR-155，miR-17-5p，miR-27a和miR-106a，它们在NSCLC病例的血清中显着降低，而miR-29c的血清中显着降低了。即使我们没有观察到等离子体的任何显着差异，并且血清中的表达水平与等离子体水平没有很好的相关性，但这些组织类型的获得方式也有所不同，这可能解释了结果的差异。此外，在所有患者中，LET-7B的血浆表达降低与癌症特异性较差的死亡率较差，而MiR-223的血清表达降低与IA/B期患者的癌症特异性死亡率相关。 Additionally, we looked at the expression of biologically relevant miRs (miR-21, miR-29b, miR-34a/b/c, miR-155, and let-7a) in formalin-fixed paraffin-embedded tumor specimens from the patients who participated in the International Adjuvant Lung Cancer Trial (IALT), the largest randomized study conducted to date of adjuvant chemotherapy in patients with radically resected NSCLC.目的是评估这些miR的表达是否代表生存的预后和预测价值。重要的是，某些miR的表达与化学抗性有关。但是，我们的结果表明，在这个大型患者队列中，所检查的miRS表达模式既没有预测性也不是预后。根据NSCLC腺癌患者肺癌标本的先前MIR微阵列数据，我们研究了5种MIR的表达，其表达在肺癌中显着改​​变，并且与癌症特异性死亡率有关。其中三个miR（miR-17，miR-21和miR-155）是潜在的致癌miR，显示出高表达水平的肿瘤表达增加与预后不良有关。我们研究了它们在来自马里兰州，挪威和日本的NSCLC患者的早期肺腺癌中的炎热组织中的表达。我们发现，在马里兰州队列中，所有三种miR的表达升高与癌症特异性死亡率较差有关。但是，只有miR-21在挪威队列中显示出相同的关联，在日本队列中，无复发的生存率较差。此外，与TNM I期患者相比，更晚期的分期肿瘤表达的miR-21水平明显更高，其中miR-21组也与癌症特异性较差的死亡率和无复发生存率相关，与其他临床因素无关。我们发现，在来自世界不同地区的3个独立的NSCLC腺癌同伴中，MiR-21表达升高与预后较差有关。这一发现具有翻译意义。 MiR-21表达具有潜在的效用，作为NSCLC腺癌患者的早期预后生物标志物。尽管我们没有观察到FFPE组织中miR-21表达的显着增加，但这可能是由于从这些组织中提取的RNA质量较低，如前所述。此外，我们采取了一种系统生物学方法来研究miR在癌症中的影响。每个mir都可以控制数百个不同的编码使者的翻译，并且一个使者可以由多个miR控制。我们结合了差异表达，遗传网络和DNA拷贝数改变。我们确认或发现了在癌症中具有全面作用的MIR：Has-Mir-103/106在癌症中被下调，而Has-Mir-30最突出。 HAS-MIR-17/92家族被放大，并在癌症中删除了Has-Mir-143/145群集；发现Has-Mir-30和Has-Mir-204在DNA拷贝数水平上进行了物理变化。我们报道了正常组织的第一个miR网络，但也建立了用于耦合癌性和非癌组织的miR网络，并确定了miR网络中的癌症变化。最后，我们将DNA变异叠加到表达数据上，以在癌症中产生全面的miR改变图。除了对miR作为肺癌重要生物标志物的研究外，我们还研究了肿瘤抑制基因LKB1/STK11，该基因具有丝氨酸 - 硫代激酶活性，位于19p13.3染色体上。 19p染色体是吸烟者肺肿瘤中第二大最常见的损失区域，在30％的肺癌细胞系和较小的原发性肺腺癌中发现了LKB1突变。在这项研究中，我们使用了几种互补的遗传方法来评估原发性NSCLC中的KLB1基因座。我们的数据确定了通过纯合缺失（HD）或杂合性丧失（LOH）对LKB1基因的总灭活，在39％的测试样品中，在39％的测试样品中，在NSCLC的90％中发生了HD或LOH丧失的HD或LOH，而在39％的测试样品中均可灭活。此外，我们研究了肿瘤雌激素受体（ERS）和血清雌激素在肺癌中的作用，假设是雌激素生物合成途径中ER和功能性单核苷酸多态性（SNP）与较差的肺癌生存有关。分析了血清雌激素，孕酮，肿瘤信使RNA的激素受体的表达和生殖系DNA多态性的与肺癌存活的关联。在这项研究中，血清雌激素第三三位数最高的患者的生存率较差。此外，雌激素受体α（ER-Alpha）多态性（rs2228480）的变异等位基因与肿瘤ER-Alpha水平升高和所有人群中较差的生存率显着相关。结果与性别和激素替代疗法无关。因此，我们报道了血清雌激素增加与肺癌男性和女性患者的生存率较差的显着关联。此外，我们研究了一种非常重要的机制，即细胞永生化，这是癌症的标志之一，依赖于端粒长度维持机制（TMM）的活性，即端粒酶或端粒的替代延长（ALT）。我们检查了43个恶性胸膜间皮瘤（MPMS）。 TMM被广泛认为是癌症疗法的潜在靶标，端粒酶抑制剂已进入临床试验。我们询问MPM使用ALT和/或端粒酶的比例。我们发现43个MPM中的43个是端粒酶阳性[+]和alt-ngative [ - ]。为了研究MPM细胞是否异常易受端粒酶激活的敏感，我们检查了在细胞永生化的体外模型中TMM的激活，其中正常胸膜间皮细胞用SV40转导。我们发现胸膜间皮瘤细胞能够在体外激活TMM，并且观察到100％mpms是端粒酶[+]表明，在这种肿瘤类型的发生期间，体内有一些因素选择用于选择端粒酶活性。我们得出的结论是，MPM是一种可以考虑用于抗细胞瘤疗法的肿瘤。我们的研究中描述的基因和机制对肺癌的起步，进展和结果很重要，在肺癌中呈现潜在的分子预后和诊断生物标志物。