Atypical Angiogenic Mimicry During Collective Lung Cancer Invasion

摘要 集体侵袭是在大多数实体肿瘤类型患者中观察到的主要转移方式。怎么了？ 集体入侵包作为一个单一的凝聚力单元进行操作、通信和导航仍不清楚。至 针对这一问题，我们在一个图像引导的基因组学平台上发表了分离集体中任何活细胞的方法(S) 入侵包装，并扩大种群进行基因组和分子分析，我们称之为 时空细胞和基因组分析(SAGA)我们用佐贺来解构集体入侵集团 并分析了作为一个等级凝聚单位入侵的领导细胞和跟随细胞的分子轮廓。至 产生集体侵袭包，先导细胞和跟随细胞经历基于血管内皮生长因子/Notch的血管生成 促进细胞的拟态程序：与血管生成相似但不完全相同的细胞协作和侵袭。 侵略型领导人分泌的血管内皮生长因子会招募大量的追随者加入集体行动；一旦行动结束 形成的先导细胞和跟随细胞经历包括DLL4拮抗剂的Notch1-DLL4细胞构图程序， 锯齿状-1(Jag1)。根据我们公布的初步数据，我们假设合作信号 在相邻细胞之间，需要通过Notch1及其配体形成空间依赖的信号事件 在入侵包里。我们认为这促进了细胞间的合作，并导致转移增加。 效率。为了测试这一点，在目标1中，我们将定义非典型的血管生成拟态如何通过Notch1/Jag1/Dll4信号 在空间上规范合作和入侵。这将是在理解 该通路如何维持集体侵袭包、驱动转移和促进细胞外基质 改建。在目标2中，我们使用Jag1作为肺癌引导细胞的生物标记物来分离第一个患者的引导细胞 并探索非典型的血管生成拟态。这使我们能够定义转移潜力和翻译影响 在肺癌患者中这一罕见但有侵袭性的人群。在整个过程中，我们利用开发的独特资源 这里包括SAGA来源的细胞系，第一组早期和晚期侵袭肺的患者来源 器官样物质，体外成像，以及一组罕见的肺原发肿瘤与转移性脑组织配对。我们 推测这些数据将提供对非典型血管生成拟态程序的机械性洞察 对了解肺癌患者先导细胞生物学的翻译价值。

Abstract Collective invasion is a major mode of metastasis observed in patients across most solid tumor types. How the collective invasion pack operates, communicates, and navigates as a single cohesive unit remains unclear. To address this, we published on an image-guided genomics platform to isolate any living cell(s) within a collective invasion pack, and expand the population for genomic and molecular analysis, a technique we termed Spatiotemporal Cellular & Genomic Analysis (SaGA). We used SaGA to deconstruct the collective invasion pack and dissect the molecular profiles of leader and follower cells invading as a hierarchical cohesive unit. To generate the collective invasion pack, leader and follower cells undergo a VEGF/Notch-based angiogenic mimicry program that promotes cell:cell cooperation and invasion that is similar, but not identical to angiogenesis. VEGF secreted by invasive leaders recruits proliferative followers into the collective pack; once the pack is formed, leader and follower cells undergo a Notch1-Dll4 cell patterning program that includes the Dll4 antagonist, Jagged-1 (Jag1). Based upon our published and preliminary data, we hypothesize that cooperative signaling among contiguous cells via Notch1 and its ligands are required to form the spatially dependent signaling events within the invasion pack. We propose that this fosters cell:cell cooperation and leads to increased metastatic efficiency. To test this, in Aim 1 we will define how atypical angiogenic mimicry via Notch1/Jag1/Dll4 signaling operates to spatially regulate cooperation and invasion. This would be a significant step forward in understanding how this pathway operates to maintain the collective invasion pack, drive metastasis, and facilitate ECM remodeling. In Aim 2, we use Jag1 as a lung cancer leader cell biomarker to isolate the first patient leader cells and probe atypical angiogenic mimicry. This allows us to define the metastatic potential and translational impact of this rare yet invasive population in lung cancer patients. Throughout, we leverage unique resources developed here including SaGA-derived cell lines, the first set of early and late-stage invading lung patient-derived organoids, ex vivo imaging, and a rare set of lung primary tumors with paired metastatic brain tissue. We speculate that these data will provide mechanistic insight into the atypical angiogenic mimicry program and translational value towards understanding lung cancer patient leader cell biology.