Project 1: Race-related RNA splicing in non-small cell lung cancer: functional interrogation and therapeutic targeting

摘要-项目1 肺癌和支气管癌是美国和我国癌症相关死亡的主要原因 该机构的北卡罗来纳州，黑人死亡人数最多，被诊断为3人 平均比白人年轻几岁。黑人非小细胞肺癌(NSCLC)的生物驱动因素 仍然没有得到充分的探索，只有少量关于种族相关的可操作突变差异的研究 和聚合基因表达。因此，这些努力很可能错过了其他重要的种族驱动力- 与NSCLC相关的生物学和临床异质性。拟议的工作解决了以下迫切需要 非小细胞肺癌中新的RACE相关RNA剪接靶点的功能特征和治疗靶点。我们是 第一个发现非洲和非洲患者之间非小细胞肺癌RNA剪接差异的团队 欧洲血统。具体地说，在肺鳞状细胞癌(LUSC)中，与种族相关的数量 差异剪接基因(DGs)(4,830)远远超过表现种族相关差异的基因数量 相同组织中的聚合基因表达(Degs)(267)。据报道，在DSG中，有17%是 癌基因、肿瘤抑制基因和/或驱动因素以及DSG内的355个RNA剪接事件是相关的 与LUSC的生存有关。据报道，在这些DEG中，6%与癌症有关，18与LUSC有关 生死存亡。许多DSG和DEG涉及治疗靶向的信号通路。此外， 我们已经挖掘了癌症基因组图谱(TCGA)，并在其他LUSC或 肺腺癌(LUAD)。拟议工作的目标是扩大这一新的调查领域 对非小细胞肺癌差异的精确肿瘤学具有重要意义：1)在扩展的 临床相关的非小细胞肺癌患者亚组的队列，注释为生存和吸烟状况，2) 在功能上表征与种族相关的备选RNA剪接事件的优先顺序，以及3)按药物优先顺序排列 用于治疗应用的种族相关的替代RNA剪接事件。为了达到这些目标，我们建议 实现三个目标。目的1：研究RNA剪接变异体和编码反式作用基因的表达 非洲和欧洲血统患者中临床相关非小细胞肺癌亚群的剪接因子。目标 2：探讨优先考虑的小种相关RNA剪接变异体的功能意义 非小细胞肺癌。目的3：a)开发新型剪接开关寡核苷酸(SSO)吗啉基类药物以调节RNA 剪接对种族相关非小细胞肺癌至关重要的事件用于治疗应用和B)以确定可用的靶向 基于异常剪接途径的抑制种族相关非小细胞肺癌的治疗药物(S)。这个 这项研究的理由和影响是：1)它将增加对分子机制的理解 潜在的肺癌差异，2)提供了大量与RNA剪接相关的新靶点 非洲血统患者非小细胞肺癌新生物标记物和治疗药物的开发，3) 结合TCGA数据，这项研究将使分子特征的非小细胞肺癌的数量增加一倍以上 来自非洲血统患者的生物样本，并产生额外的临床前模型，制作这样的数据 以及可供全国进行肺癌和肺癌研究的科学家队列使用的模型 差异，以及4)在非洲血统患者中用于非小细胞肺癌的新型RNA剪接靶向药物的位置 活体研究。最终，这种精确的肿瘤学干预和进一步的研究是由分子水平的 具有特征的生物菌群和模型将有可能缓解非小细胞肺癌的差异。

ABSTRACT – Project 1 Lung and bronchus cancer is the leading cause of cancer-related deaths in the United States and in our institution’s state of North Carolina, with blacks having the highest number of deaths and being diagnosed three years younger on average than whites. Biological drivers of non-small cell lung cancer (NSCLC) in black patients remain underexplored, with only a small number of studies on race-related differences in actionable mutations and aggregate gene expression. As a result, these efforts have likely missed other important drivers of race- related NSCLC biological and clinical heterogeneity. The proposed work addresses the urgent need to functionally characterize and therapeutically target novel race-related RNA splicing targets in NSCLC. We are the first team to identify alternative RNA splicing differences in NSCLC between patients of African and European ancestry. Specifically, in lung squamous cell carcinomas (LUSCs), the number of race-related differentially spliced genes (DSGs) (4,830) far exceeded the number of genes exhibiting race-related differential aggregate gene expression (DEGs) (267) in the same tissues. Among the DSGs, 17% are reported to be oncogenes, tumor suppressor genes and/or drivers and 355 RNA splicing events within DSGs are associated with LUSC survival. Among the DEGs, 6% are reported to be cancer-related and 18 are associated with LUSC survival. A number of the DSGs and DEGs involve therapeutically targetable signaling pathways. Furthermore, we have mined The Cancer Genome Atlas (TCGA) and have identified DSGs and DEGs in additional LUSCs or lung adenocarcinomas (LUADs). The objectives of the proposed work are to extend this novel area of inquiry with significance for precision oncology in NSCLC disparities by 1) examining DSGs and DEGs in an expanded cohort of clinically relevant NSCLC subgroups of patients annotated for survival and smoking status, 2) functionally characterizing prioritized race-related alternative RNA splicing events, and 3) drugging prioritized race-related alternative RNA splicing events for therapeutic application. To reach these objectives, we propose to conduct three aims. Aim 1: To assess the expression of RNA splice variants and genes encoding trans-acting splicing factors across clinically relevant NSCLC subgroups in patients of African and European ancestry. Aim 2: To interrogate the functional significance of prioritized race-related RNA splice variants for the biology of NSCLC. Aim 3: A) To develop novel splice-switching oligonucleotide (SSO) morpholino drugs to modulate RNA splicing events critical to race-related NSCLC for therapeutic application and B) To identify available targeted therapeutic agents that inhibit race-related NSCLC based on dysregulated RNA splicing pathway(s). The rationale for and impact of this study is that it will 1) increase understanding of the molecular mechanisms underlying lung cancer disparities, 2) provide an abundance of novel RNA splicing-related targets for development of new biomarkers and therapeutic agents for NSCLC in patients of African ancestry, 3) when combined with TCGA data, this study will more than double the number of molecularly characterized NSCLC biospecimens from patients of African ancestry, and generate additional preclinical models, making such data and models available to the nationwide cohort of scientists conducting research on lung cancer and lung cancer disparities, and 4) position lead novel RNA splicing-targeted drugs for NSCLC in patients of African ancestry for in vivo studies. Ultimately, such precision oncology interventions and further studies enabled by the molecularly characterized cohort of biospecimens and models will have the potential to mitigate NSCLC disparities.