Identification of a novel targetable cancer stem cell regulator promoting cancer progression and metastasis in non-small cell lung cancer

非小细胞肺癌（NSCLC）占所有肺癌的85%左右。的 5- 年生存率约为 IIIA期NSCLC为14%， IIIB期为5%。然而一旦 NSCLC已达到IV期 并且转移到不同的地方，这是非常困难的治疗。 5- IV期年生存率 NSCLC是 大约1%。靶向治疗如抗EGFR或抗ALK是晚期NSCLC的一线治疗 EGFR或ALK突变，而铂类化疗是晚期 无靶向突变的NSCLC。有趣的是，最近的研究表明，抗PD 1/PDL 1免疫疗法 是治疗晚期NSCLC的一种新的有效策略。虽然NSCLC患者最初表现出极大的受益， 治疗时，反应只是短暂的，持续时间相对较短，可能是由于获得耐药性， 机制等因此，迫切需要确定新的和有效的治疗策略， NSCLC伴转移。具有CSC特性的小细胞群有助于癌症的发生、发展 和转移以及各种癌症（如NSCLC）的耐药性，但消除 目前缺乏CSC，代表CSC和NSCLC靶向的临床需求未得到满足。虽然CSC拥有 由于免疫逃逸特性，目前尚不清楚非CSC癌细胞如何占癌细胞总数的大部分 种群也可以抵抗免疫细胞的攻击。这项研究的目的是描述一种新颖独特的 NSCLC中的CSC群体及其调控机制，可用于开发新的有效的 治疗晚期NSCLC和/或克服对当前标准治疗的耐药性的策略。我们的研究 鉴定了一种位于细胞膜中的新型可药用调节剂，用于维持CSC、癌症进展和 NSCLC的转移及其过表达预示着NSCLC患者的不良生存结局。遗传上或 靶向这一新发现的调节因子可减弱维持CSC的致癌信号 这些特性和免疫逃逸导致NSCLC的癌症进展和转移。我们假设 在癌症中存在的具有CSC特性的独特细胞群可以传递致癌和免疫逃逸 向非CSC癌细胞发出信号，从而赋予大量癌细胞免疫逃逸特性。我们提出 三个具体目标，这是高度支持我们的创新初步结果，以进一步表征 这种新的调节剂及其配体的作用和潜在机制以及它们在调节中的靶向， CSC、NSCLC的进展和转移。我们的建议具有高度的独创性和重要意义， 一个突破性的概念，确定了一种具有CSC和免疫逃逸特性的新型检查点阻断剂， 利用切割技术，包括无偏转录组学，Cas9/CRISPR编辑，患者来源的 类器官、患者来源的异种移植（PDX）模型以及人源化小鼠和遗传敲入小鼠模型 来验证我们的假设和概念我们的研究彻底改变了， 显著地推进了我们对CSC和癌症免疫调节的理解，但也提供了一个新的范例， 和针对晚期NSCLC的策略。

Non-small cell lung cancer (NSCLC) accounts for around 85% of all lung cancer. The 5- year survival rate is about 14% for stage IIIA NSCLC, while it is about 5% for stage IIIB. However, once NSCLC has reached to the stage IV and metastasized to different places, it is very difficult to treat. The 5- year survival rate for stage IV NSCLC is just about 1%. Targeted therapy such as anti-EGFR or anti-ALK is the frontline treatment for advanced NSCLC with EGFR or ALK mutations, while platinum-based chemotherapy is the first line treatment for advanced NSCLC without targetable mutations. Interestingly, recent studies suggest that anti-PD1/PDL1 immunotherapy is a new and effective strategy for advanced NSCLC. While NSCLC patients initially show great benefit from these treatments, the response is only transient with relatively short duration likely due to acquiring resistant mechanisms. Identification of novel and effective therapeutic strategies is therefore an urgent need for advanced NSCLC with metastasis. A small cell population with CSC properties contributes to cancer initiation, progression and metastasis as well as drug resistance in various cancers such as NSCLC, but an effective strategy to eliminate CSCs is currently lacking, representing an unmet clinical need for CSC and NSCLC targeting. While CSCs possess immune escape properties, it is unclear how non-CSC cancer cells accounting for the majority of total cancer cell populations could also resist from immune cell attack. The goal of this study is to characterize a novel and unique CSC population in NSCLC and its regulatory mechanisms that can be harnessed for developing a novel effective strategy for advanced NSCLC and/or for overcoming the resistance to current standard of care. Our study identifies a novel druggable regulator localized in cell membrane for maintaining CSCs, cancer progression and metastasis of NSCLCs and its overexpression predicts poor survival outcome NSCLC patients. Genetically or pharmacologically targeting this newly identified regulator attenuates oncogenic signal for maintain CSC properties and immune escape leading to cancer progression and metastasis of NSCLC. We hypothesized that a unique cell population with CSC properties existed in cancer can transmit an oncogenic and immune escape signal to non-CSC cancer cells, thereby endowing bulk cancer cells with immune escape properties. We proposed three specific aims, which are highly supported by our innovative preliminary results, to further characterize the roles and underlying mechanisms of this novel regulator and its ligand as well as their targeting in regulating CSCs, progression and metastasis of NSCLC. Our proposal is highly original and significant, as we have proposed a breakthrough concept, identified a novel checkpoint blocker with CSC and immune escape properties and utilized cutting technologies including unbiased transcriptomics, Cas9/CRISPR editing, patient-derived organoids, patient derived xenograft (PDX) models, and humanized mice and genetic knockin mouse models with intact immunity to validate our provocative hypothesis and concept. Our study has revolutionized and significantly advanced our understanding of CSC and cancer-immune regulation, but also offers a new paradigm and strategy for targeting advanced NSCLC.