PROGNOSTIC SIGNIFICANCE AND ANALYSIS OF iNOS IN MELANOMA

黑色素瘤中 iNOS 的预后意义和分析

• 批准号: 6993428
• 负责人: ELIZABETH A GRIMM
• 金额: $ 15.74万
• 依托单位: UNIVERSITY OF TX MD ANDERSON CAN CTR
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-07-09 至 2009-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6993428
• 关键词: biomarker; enzyme activity; enzyme induction /repression; gene mutation; genetic transcription; human subject; human tissue; melanoma; metastasis; mitogen activated protein kinase; neoplasm /cancer classification /staging; neoplasm /cancer genetics; neoplastic process; nitric oxide synthase; nuclear factor kappa beta; patient oriented research; prognosis; protein quantitation /detection; protooncogene; tyrosine

Current data suggest that the process of melanoma tumorigenesis is enhanced by aberrant constitutive expression of the inducible form of nitric oxide synthase (iNOS), an enzyme catalyzing the generation of nitric oxide (NO), normally in response to cytokines. The presence of reactive NO in melanoma is indicated by detection of nitrotyrosine (NT), which is an end-product of the chemical reaction of NO with proteins; NT formation is known to alter intracellular signaling, especially when the target is a tyrosine kinase substrate. Production of low levels (pM) of NO by human tumors is proposed to be responsible for increased angiogenesis, growth, invasion, genomic instability and resistance to apoptosis. Our analysis of human melanoma specimens from patients with advanced (Stage III) disease indicates that patients whose tumors express iNOS are more likely to die within 2 years of neoadjuvant biohemotherapy than patients without iNOS (p<0.001). Specific Aim 1 comes from the clinic to the laboratory to further test the significance of iNOS and NT in tissue specimens as prognostic markers for primary and metastatic melanoma patients. The translational goals of Specific Aim 1 are validation of iNOS and NT as new prognostic markers for survival in melanoma patients and to determine whether either marker provides independent prognostic value. Significant results will then be submitted to American Joint Commission on Cancer, Committee on Melanoma for inclusion in future staging revisions, as the first molecular marker for melanoma to be validated. Specific Aim 2 tests the hypothesis that iNOS is controlled by specific, identifiable gene transcriptional regulators including constitutively active NFKB. NFKB activation is known to regulate basal as well as cytokine-induced iNOS in normal human cells, however its role in regulation of melanoma iNOS is unknown. Specific Aim 3 builds on further upstream mechanisms of iNOS activation and tests the association and functional role of mutated B-raf, which is reported to drive MAPK independently of Ras in melanoma cells. A novel molecular heteroduplex analysis will be employed to detect mutated B-raf in paraffin-embedded tumor specimens, and study of its association with iNOS and constitutive NFKappaB. The regulation of mutated B-raf activity is also proposed in Specific Aim 4 as a means to regulate iNOS and, by extension, melanoma growth.

当前的数据表明，黑色素瘤肿瘤发生的过程通过诱导型一氧化氮合酶（INOS）（INOS）的异常表达增强，这是一种催化一氧化氮（NO）的酶，通常是响应细胞因子而产生的。黑色素瘤中反应性NO的存在是通过检测硝基酪氨酸（NT）来指示的，硝基酪氨酸（NT）是NO与蛋白质的化学反应的最终产物。已知NT形成会改变细胞内信号传导，尤其是当靶标是酪氨酸激酶底物时。 有人认为人类肿瘤的低水平（PM）的生产是为了增加血管生成，生长，侵袭，基因组不稳定性和对凋亡的抗性。我们对患有晚期（III期）疾病患者的人类黑色素瘤标本的分析表明，与没有iNOS的患者相比，在新辅助生物学疗法的2年内，表达iNOS的患者更有可能在两年内死亡（p <0.001）。具体目标1来自诊所到实验室，以进一步测试INOS和NT在组织样品中的重要性，作为原发性和转移性黑色素瘤患者的预后标记。 特定目标1的翻译目标是对黑色素瘤患者生存的新预后标记的验证，并确定任何一个标记物是否提供独立的预后价值。然后，重大结果将提交给美国癌症联合委员会，黑色素瘤委员会，以纳入将来的分期修订，这是对黑色素瘤的第一个分子标记。具体目标2检验了iNOS由特定的，可识别的基因转录调节因子（包括组成型活性NFKB）控制的假设。已知NFKB激活在正常人细胞中调节基础和细胞因子诱导的iNOS，但是其在调节黑色素瘤iNOS中的作用尚不清楚。特定的目标3建立在iNOS激活的进一步上游机制上，并测试突变的B-RAF的关联和功能作用，据报道，这是独立于RAS驱动MAPK的RAS。 黑色素瘤细胞。将采用一种新型的分子杂化分析来检测石蜡包含的肿瘤标本中的突变B-RAF，并研究其与INOS和组成型NFKappab的关联。在特定目标4中还提出了突变的B-RAF活性的调节，作为调节iNOS并扩展黑色素瘤生长的一种手段。