肺癌仍然是美国和全世界癌症死亡的主要原因。非小细胞肺癌 (NSCLC) 具有高转录组瘤内异质性 (RNA-ITH)，限制了基于表达的预后模型的可重复性。在这项研究中，我们使用来自公共 (TRACERx) 和内部 (MDAMPLC) 队列的多区域 RNA-seq 数据（来自 350 个人的 880 个肿瘤样本）来研究 RNA-ITH 对局部 NSCLC 预后的影响，包括基因、特征、和肿瘤微环境水平。在基因水平上，肿瘤内不同区域危险基因的最大表达（与生存呈负相关的表达）和保护性基因的最小表达（与生存呈正相关的表达）比平均表达更具预后性。接下来，我们检查了多区域表达谱是否可以提高预后特征的性能。我们调查了从之前出版物中收集的 11 个基因特征以及本研究中开发的一个特征。对于所有这些，通过使用简单的函数将特征基因的区域表达转换为样本特异性表达——取危险基因的最大表达量和保护性基因的最小表达量，可以显着提高预后预测的准确性。在肿瘤微环境中，我们发现类似的规则似乎也适用于免疫ITH。我们根据表达反卷积计算了样本每个区域中主要免疫细胞类型的浸润水平。预后分析表明，保护性免疫细胞浸润水平最低或有害免疫细胞浸润水平最高的区域决定了NSCLC患者的预后。我们的研究强调了 RNA-ITH 对 NSCLC 预后的影响，应考虑到这一影响，以优化基于表达的预后生物标志物和模型的设计和应用。多区域检测具有显着改善其预后分层应用的巨大潜力。© 2024。作者。

Lung Cancer remains the leading cause of cancer deaths in the USA and worldwide. Non-small cell lung cancer (NSCLC) harbors high transcriptomic intratumor heterogeneity (RNA-ITH) that limits the reproducibility of expression-based prognostic models. In this study, we used multiregional RNA-seq data (880 tumor samples from 350 individuals) from both public (TRACERx) and internal (MDAMPLC) cohorts to investigate the effect of RNA-ITH on prognosis in localized NSCLC at the gene, signature, and tumor microenvironment levels. At the gene level, the maximal expression of hazardous genes (expression negatively associated with survival) but the minimal expression of protective genes (expression positively associated with survival) across different regions within a tumor were more prognostic than the average expression. Following that, we examined whether multiregional expression profiling can improve the performance of prognostic signatures. We investigated 11 gene signatures collected from previous publications and one signature developed in this study. For all of them, the prognostic prediction accuracy can be significantly improved by converting the regional expression of signature genes into sample-specific expression with a simple function-taking the maximal expression of hazardous genes and the minimal expression of protective genes. In the tumor microenvironment, we found a similar rule also seems applicable to immune ITH. We calculated the infiltration levels of major immune cell types in each region of a sample based on expression deconvolution. Prognostic analysis indicated that the region with the lowest infiltration level of protective or highest infiltration level of hazardous immune cells determined the prognosis of NSCLC patients. Our study highlighted the impact of RNA-ITH on the prognostication of NSCLC, which should be taken into consideration to optimize the design and application of expression-based prognostic biomarkers and models. Multiregional assays have the great potential to significantly improve their applications to prognostic stratification.© 2024. The Author(s).