PQ 5 Exploring whole mitochondrial genome of screen detected pre-neoplastic lung lesions by novel approaches

项目总结： 人类线粒体基因组(MG)容易发生基因改变，导致体细胞突变， 线粒体DNA(MtDNA)拷贝数和呼吸链亚基表达改变。这 部分原因是线粒体DNA在受损时自我修复的能力有限。虽然这是一场集结 线粒体DNA的体细胞突变与癌症风险的增加有关，线粒体DNA改变在 肺腺癌(LA)的恶变尚未得到解决。因此，一个明确的 了解mtDNA在肺肿瘤发生过程中的变化是鉴定分子的关键 与癌症发生和肿瘤进展相关的生物标志物。 像所有实体肿瘤一样，LA被认为是通过一系列基因改变而启动和进展的， 包括线粒体DNA的变化。因原发性腺癌而切除的肺通常有微小的离散病灶。 细胞学上不典型的肺细胞增殖称为非典型腺瘤性增生(AAH)。 有证据表明，AAH是发展为原位腺癌(AIS)的第一步， 微侵袭性腺癌(MIA)和最终完全侵袭性腺癌。 为了描绘线粒体克隆性异质性与肿瘤进展的关系，我们将使用新的高密度的 吞吐量-基于PCR的浓缩技术与下一代测序(NGS)相结合，以评估 原发侵袭性AAH患者皮损标本线粒体DNA的变化 腺癌或来自同一AIS和MIA内不同组织进展区。通过这种方式，我们 将对早期病变的线粒体异质性及其潜在作用有一个独特的理解 早期腺瘤进展中的克隆性事件。接下来，我们将对检测到的LA进行排序 通过螺旋CT筛查和间歇性癌，可能表明其进展迅速 表型。最后，我们将在配对血浆样本中验证线粒体DNA衍生突变 稳健的数字聚合酶链式反应方法，为非侵入性早期肺癌检测提供了新的机会。 综上所述，这项建议将确定线粒体DNA来源的体细胞突变沿着LA和 将开发生物标记物，使我们能够更好地非侵入性地预测早期病变的命运。

Project Summary: The human mitochondrial genome (MG) is prone to genetic alterations, which lead to somatic mutations, changes in mitochondrial DNA (mtDNA) copy number and altered expression of respiratory chain subunits. This is in part due to the fact that mtDNA has a limited ability to repair itself when it is damaged. Although a buildup of somatic mutations in mtDNA has been associated with increased cancer risk, the role of mtDNA alterations in malignant transformation of lung adenocarcinoma (LA) has not been addressed. Therefore, a clear understanding of changes in mtDNA along the pathway of lung tumorigenesis is critical for identifying molecular biomarkers related to carcinogenesis and tumor progression. Like all solid tumors, LA is thought to be initiated and to progress through a series of genetic alterations, including changes in mtDNA. Lungs resected for primary adenocarcinomas often harbor minute discrete foci of cytologically atypical pneumocyte proliferations designated as atypical adenomatous hyperplasia (AAH). Evidence suggests that AAH represents an initial step in the progression to adenocarcinoma in situ (AIS), minimally invasive adenocarcinoma (MIA) and ultimately fully invasive adenocarcinoma. To delineate mitochondrial clonal heterogeneity as a function of tumor progression we will use novel high- throughput-PCR-based enrichment technology combined with next generation sequencing (NGS) to assess mtDNA alterations in samples isolated from AAH lesions collected from patients diagnosed with primary invasive adenocarcinoma or from different zones of histologic progression within the same AIS and MIA. In this way, we will gain a unique understanding of the mitochondrial heterogeneity of early lesions and the potential role of these clonal events in the progression of early glandular neoplasms. Next, we will sequence LA that were detected through spiral CT screening approach and interval cancers, which likely indicate their rapidly progressing phenotypes. Finally, we will validate mtDNA-derived mutations in paired plasma samples using a sensitive and robust digital PCR approach, which provides a novel opportunity for noninvasive early lung cancer detection. In summary, this proposal will identify mtDNA-derived somatic mutations along the progression of LA and will develop biomarkers that will allow us to better predict the fate of early lesions non-invasively.