Atypical Angiogenic Mimicry During Collective Lung Cancer Invasion

摘要 集体侵袭是在大多数实体瘤类型的患者中观察到的主要转移模式。如何 集体入侵包作为一个单一的内聚单位运作，通信和导航仍然不清楚。到 为了解决这个问题，我们发表了一个图像引导的基因组学平台，以分离集体中的任何活细胞。 入侵包装，并扩大人口的基因组和分子分析，一种技术，我们称之为 时空细胞和基因组分析（佐贺）。我们用佐贺来解构集体入侵包 并将入侵的领导细胞和跟随细胞作为一个层次性的凝聚单位进行分子剖析。到 产生集体入侵包，领导和追随者细胞经历基于VEGF/Notch的血管生成 模拟程序，促进细胞：细胞的合作和入侵，类似，但不完全相同的血管生成。 由入侵性领导者分泌的VEGF将增殖性追随者招募到集体包中;一旦包 形成的前导细胞和跟随细胞经历包括D11 4拮抗剂的Notch 1-D11 4细胞图案化程序， 锯齿状-1（Jag 1）。基于我们发表的和初步的数据，我们假设合作信号 在相邻细胞之间通过Notch 1及其配体是形成空间依赖性信号事件所必需的 在入侵包中。我们认为，这促进了细胞：细胞合作，并导致增加转移 效率为了验证这一点，在目标1中，我们将定义非典型血管生成拟态如何通过Notch 1/Jag 1/Dll 4信号传导 在空间上调节合作和入侵。这将是我们了解 这一通路如何运作以维持集体侵袭包，驱动转移，并促进ECM 重塑在目标2中，我们使用Jag 1作为肺癌前导细胞生物标志物来分离第一个患者前导细胞。 探索非典型血管生成拟态这使我们能够确定转移潜力和翻译影响， 这种罕见但具有侵袭性的人群的肺癌患者。在整个过程中，我们利用开发的独特资源 这里包括SaGA衍生的细胞系，第一组早期和晚期侵袭性肺患者衍生的细胞系， 类器官、离体成像和一组罕见的肺原发性肿瘤与成对的转移性脑组织。我们 推测这些数据将为非典型血管生成拟态程序提供机制上的见解， 对理解肺癌患者领导细胞生物学的转化价值。

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.