Abstract：
BACKGROUND: It was highlighted by recent studies on the biological significance of vasculogenic mimicry (VM) in tumorigenicity and progression. However, it is unclear whether VM also plays a potential role in immune regulation and tumor microenvironment (TME) formation.
METHODS: To identify patterns of VM alterations and VM-associated genetic features in non-small cell lung adenocarcinoma, we have screened 309 VM regulators and performed consensus molecular typing by the NMF algorithm. The ssGSEA and CIBORSORT algorithms were employed to measure the relative infiltration of distinct immune cell subpopulations. Individual tumors with immune responses were evaluated for alteration patterns of VM with typing-based differential genes.
RESULTS: In 490 LUAD samples, two distinctive VM alteration patterns connected to different clinical outcomes and biochemical pathways were established. TME characterization showed that the observed VM patterns were primarily saturated with cell proliferation and metabolic pathways and higher in immune cell infiltration of the C1 type. Vasculogenic mimicry-related genes (VMRG) risk scores were constructed to divide patients with lung adenocarcinoma into subgroups with high and low scores. Patients with lower scores had better immunological scores and longer survival times. Upon further investigation, higher scores were positively correlated with higher tumor mutation burden (TMB), M1-type macrophages and immune checkpoint molecules. Nevertheless, in two other immunotherapy cohorts, individuals with lower scores had enhanced immune responses and long-lasting therapeutic benefits. Finally, we monitored the ANLN gene from the VMRG model, which was highly expressed in lung adenocarcinoma tissues and negatively correlated with prognosis; it was also highly expressed in lung adenocarcinoma cell lines, and knockdown of ANLN elicited low expression of VEGFA, MMP2 and MMP9.
CONCLUSIONS: This study highlights that VM modifications are significantly associated with the diversity and complexity of TME, revealing new features of the immune microenvironment in lung adenocarcinoma and providing a new strategy for immunotherapy. Screening ANLN as a critical target for vasculogenic mimicry in lung adenocarcinoma provides a novel perspective for the targeted treatment of lung adenocarcinoma.
METHODS: To identify patterns of VM alterations and VM-associated genetic features in non-small cell lung adenocarcinoma, we have screened 309 VM regulators and performed consensus molecular typing by the NMF algorithm. The ssGSEA and CIBORSORT algorithms were employed to measure the relative infiltration of distinct immune cell subpopulations. Individual tumors with immune responses were evaluated for alteration patterns of VM with typing-based differential genes.
RESULTS: In 490 LUAD samples, two distinctive VM alteration patterns connected to different clinical outcomes and biochemical pathways were established. TME characterization showed that the observed VM patterns were primarily saturated with cell proliferation and metabolic pathways and higher in immune cell infiltration of the C1 type. Vasculogenic mimicry-related genes (VMRG) risk scores were constructed to divide patients with lung adenocarcinoma into subgroups with high and low scores. Patients with lower scores had better immunological scores and longer survival times. Upon further investigation, higher scores were positively correlated with higher tumor mutation burden (TMB), M1-type macrophages and immune checkpoint molecules. Nevertheless, in two other immunotherapy cohorts, individuals with lower scores had enhanced immune responses and long-lasting therapeutic benefits. Finally, we monitored the ANLN gene from the VMRG model, which was highly expressed in lung adenocarcinoma tissues and negatively correlated with prognosis; it was also highly expressed in lung adenocarcinoma cell lines, and knockdown of ANLN elicited low expression of VEGFA, MMP2 and MMP9.
CONCLUSIONS: This study highlights that VM modifications are significantly associated with the diversity and complexity of TME, revealing new features of the immune microenvironment in lung adenocarcinoma and providing a new strategy for immunotherapy. Screening ANLN as a critical target for vasculogenic mimicry in lung adenocarcinoma provides a novel perspective for the targeted treatment of lung adenocarcinoma.
摘要：
背景：最近关于血管生成拟态（VM）在致瘤性和进展中的生物学意义的研究强调了这一点。然而,目前尚不清楚VM是否也在免疫调节和肿瘤微环境(TME)形成中发挥潜在作用.
方法：为了确定非小细胞肺腺癌中VM改变的模式和VM相关的遗传特征，我们已经筛选了309个VM调节器，并通过NMF算法进行了一致分子分型。ssGSEA和CIBORSORT算法用于测量不同免疫细胞亚群的相对浸润。评估具有免疫应答的个体肿瘤的具有基于分型的差异基因的VM的改变模式。
结果：在490个LUAD样本中，我们建立了与不同临床结局和生化通路相关的两种独特的VM改变模式.TME表征表明，观察到的VM模式主要被细胞增殖和代谢途径饱和，而C1型免疫细胞浸润更高。构建血管生成拟态相关基因(VMRG)风险评分,将肺腺癌患者分为得分高、得分低的亚组。得分较低的患者具有更好的免疫学评分和更长的生存时间。经进一步调查,较高的分数与较高的肿瘤突变负荷（TMB）呈正相关，M1型巨噬细胞和免疫检查点分子。然而，在另外两个免疫疗法队列中，得分较低的个体具有增强的免疫应答和持久的治疗获益.最后,我们从VMRG模型监测ANLN基因，在肺腺癌组织中高表达,与预后呈负相关;在肺腺癌细胞株中高表达,ANLN的敲除引起VEGFA的低表达，MMP2和MMP9。
结论：本研究强调VM修饰与TME的多样性和复杂性显著相关。揭示了肺腺癌免疫微环境的新特征，为免疫治疗提供了新的策略。筛选ANLN作为肺腺癌血管生成拟态的关键靶点，为肺腺癌的靶向治疗提供了新的视角。
方法：为了确定非小细胞肺腺癌中VM改变的模式和VM相关的遗传特征，我们已经筛选了309个VM调节器，并通过NMF算法进行了一致分子分型。ssGSEA和CIBORSORT算法用于测量不同免疫细胞亚群的相对浸润。评估具有免疫应答的个体肿瘤的具有基于分型的差异基因的VM的改变模式。
结果：在490个LUAD样本中，我们建立了与不同临床结局和生化通路相关的两种独特的VM改变模式.TME表征表明，观察到的VM模式主要被细胞增殖和代谢途径饱和，而C1型免疫细胞浸润更高。构建血管生成拟态相关基因(VMRG)风险评分,将肺腺癌患者分为得分高、得分低的亚组。得分较低的患者具有更好的免疫学评分和更长的生存时间。经进一步调查,较高的分数与较高的肿瘤突变负荷（TMB）呈正相关，M1型巨噬细胞和免疫检查点分子。然而，在另外两个免疫疗法队列中，得分较低的个体具有增强的免疫应答和持久的治疗获益.最后,我们从VMRG模型监测ANLN基因，在肺腺癌组织中高表达,与预后呈负相关;在肺腺癌细胞株中高表达,ANLN的敲除引起VEGFA的低表达，MMP2和MMP9。
结论：本研究强调VM修饰与TME的多样性和复杂性显著相关。揭示了肺腺癌免疫微环境的新特征，为免疫治疗提供了新的策略。筛选ANLN作为肺腺癌血管生成拟态的关键靶点，为肺腺癌的靶向治疗提供了新的视角。
