Lung epithelial lineage-specific factors in the control of immune system evasion genes in tumor cells

摘要 能够逃脱宿主免疫系统防御的癌细胞通常采取更具侵略性的 行为。针对这些机制的新型免疫疗法现已成功地应用于治疗。 在许多癌症中，包括高度侵袭性的肺癌。然而，这些疗法并不能限制癌症。 在所有患者中都有进展，强调需要更有效的免疫调节药物。最终， 然而，为了开发这些新的治疗方法，科学家们首先需要了解癌症 细胞控制免疫逃避基因的表达，也称为免疫检查点。特定于家族的 在器官发育过程中决定细胞命运的转录因子在癌症中也发挥着谱系功能-- 特定的肿瘤驱动因素。在肺中，谱系特异的转录因子NKX2-1(TTF1)，这是关键的 上皮细胞发育，在肺癌中显示出促肿瘤和抗肿瘤活性，取决于细胞环境。 此外，它在癌细胞中的表达已经被证明塑造了肿瘤免疫微环境。我们 最近报道了NKX2-1-AS1，一种覆盖在NKX2-1启动子上的反义长非编码RNA， 在人胎肺上皮细胞和成人肺上皮细胞中大量表达，其水平在肺中进一步升高。 癌症，符合NKX2-1模式。此外，我们还发现NKX2-1和NKX2-1-AS1的相对水平存在差异 NKX2-1和NKX2-1-AS1具有相反的作用 免疫检查点基因PD-L1的表达。在初步分析中，我们发现NKX2-1 而NKX2-1AS1对免疫系统中的其他各种基因的控制也有相反的作用 逃避。这些结果突显了肺癌细胞平衡宿主免疫的新机制。 反应，同时控制固有的癌细胞特性。基于这些发现，我们假设肺 癌细胞通过肺上皮细胞系特异性因子NKX2-1和NKX2-1-AS1控制多个 免疫检查点基因和癌细胞逃避免疫系统的攻击。我们将对此进行测试 假设有两个目的。在目标1中，我们将在体外确定NKX2-1-AS1限制的分子机制 肺癌细胞系免疫逃逸相关基因的表达及其与免疫系统的相互作用 Nkx2-1和其他关键转录因子，如STAT3。在目标2中，我们将使用共培养系统测试 不同上皮细胞NKX2-1-AS1和NKX2-1表达比例对T细胞活化的功能影响 这些研究有可能对肺部医学领域产生重大影响，并改变 我们了解了肺上皮细胞系如何控制肺免疫反应，奠定了基础 寻找新的更有效的抗癌疗法。

ABSTRACT Cancer cells that are capable of escaping host immune system defenses typically adopt more aggressive behaviors. Novel immunotherapies targeting these mechanisms are now successfully employed in the treatment of many cancers, including highly aggressive forms of lung cancer. However, these therapies do not limit cancer progression in all patients, emphasizing a need for more effective immune-modulating drugs. Ultimately, however, to develop these new treatments, scientists first need to understand the mechanisms by which cancer cells control the expression of immune evasion genes, also called immune checkpoints. Lineage-specific transcription factors that establish cell fate during organ development also play functions in cancer as lineage- specific tumor drivers. In the lung, the lineage-specific transcription factor NKX2-1 (TTF1), which is critical for epithelial development, shows pro- and anti-oncogenic activity in lung cancer depending on cell context. Moreover, its expression in cancer cells has been shown to shape the tumor immune microenvironment. We have recently reported that NKX2-1-AS1, an antisense long noncoding RNA overlapping the NKX2-1 promoter, is abundantly expressed in human fetal and adult lung epithelial cells, and its levels are further increased in lung cancer, matching NKX2-1 patterns. Furthermore, we found that relative levels of NKX2-1 and NKX2-1-AS1 vary considerably among different cancer cell populations and that NKX2-1 and NKX2-1-AS1 have opposing effects on the expression of the immune checkpoint gene PD-L1. In preliminary analysis, we discovered that NKX2-1 and NKX2-1AS1 also have opposing effects on the control of various other genes involved in immune system evasion. These results highlight a novel mechanism by which lung cancer cells can balance host-immune responses while controlling intrinsic cancer cell properties. Based on these findings, we hypothesize that lung cancer cells, through the lung epithelial lineage-specific factors NKX2-1 and NKX2-1-AS1, control multiple immune checkpoint genes and the escape of cancer cells from the immune system attack. We will test this hypothesis in two Aims. In Aim 1, we will define in vitro the molecular mechanism by which NKX2-1-AS1 limits the expression of genes involved in immune system evasion in lung cancer cell lines through the interaction with NKX2-1 and other key transcription factors such as STAT3. In Aim 2, we will test, using a co-culture system, the functional effect of different epithelial NKX2-1-AS1 and NKX2-1 expression ratios on the activation of T-cells. These studies have the potential to impact the field of pulmonary medicine significantly and transform the way we understand how lung epithelial-lineage factors control pulmonary immune responses, laying the foundation for new and more effective anti-cancer therapies.