喉癌血管生成拟态的超声分子影像研究

Vasculogenic Mimicry (VM), is a novel type of blood supplement involved in the development, metastasis and recurrence of the tumor. It was identified by the formation of a tubular, fracture-like structure by tumor cells but not endothelial cells. Targeted contrast-enhanced ultrasound imaging is increasingly being recognized as a powerful imaging strategy for the early detecting and monitoring tumor angiogenesis, atherosclerosis, inflammation, and thrombus at the molecular level. Our preliminary studies have constructed a RGD microbubbles (MBs) that can specifically binding to αvβ3 integrin-expressing cells via biotin-avidin linkage. Moreover, these RGD-MBs enable noninvasive in vivo visualization of changes in tumor angiogenesis during tumor growth in subcutaneous cancer xenografts. In this project, we propose preparation of a novel microbubbles targeted to vasculogenic mimicry, which has been shown to be expressing in laryngeal carcinoma. In Aim 1 of this proposal, we will identify the matrix metalloproteinases-1 (MMP -1)-targeted microbubbles have a high specificity for binding to VM over expressing cells of laryngeal carcinoma. In Aim 2, in different stage of human laryngeal carcinoma (Hep-2) xenografts, we will evaluate the role of VM in laryngeal carcinoma, as well as the correlation between VM and tumor metastasis, invasion, and recurrence by using targeted ultrasound molecular imaging.

血管生成拟态(VM)是一种不依赖宿主血管内皮细胞的全新的微循环模式，与肿瘤的生长、转移及复发密切相关。如何利用影像学手段从分子水平阐明VM的潜在机制，成为现代医学影像学的重要课题。靶向超声分子显像目前已成功应用于肿瘤、动脉粥样硬化、炎症、血栓等多种疾病的早期诊断及病程监测，但迄今未见其用于评价喉癌血管生成拟态的报道，VM与喉癌浸润转移的内在联系仍不明确。我们的前期研究以“生物素 - 亲和素”体系成功制备出新型的能与血管内皮细胞特异性黏附的RGD靶向微泡造影剂。进一步地，将RGD靶向微泡用于移植瘤动物模型体内超声分子成像实验，结果显示超声分子显像可有效评价整合素 ανβ3在肿瘤血管新生中的表达。基于以上研究基础，本研究拟制备一种新型的靶向肿瘤血管生成拟态的基质金属蛋白酶-1微泡造影剂，建立不同生长阶段裸鼠人喉癌移植瘤动物模型，从超声分子影像学水平探讨VM形成与喉癌浸润转移的关系及潜在机制。

本研究拟制备一种新型的靶向肿瘤血管生成拟态的基质金属蛋白酶-1微泡造影剂，建立不同生长阶段裸鼠人喉癌移植瘤动物模型，从超声分子影像学水平探讨VM形成与喉癌浸润转移的关系及潜在机制。.目前本研究项目已按计划顺利完成。.① 课题组已完成靶向MMP-1超声微泡的制备和相应理化性质等指标的检测，检测了靶向微泡的体外超声成像性能鉴定；.② MMP-1靶向微泡和IgG MBs对照组微泡对HEp-2的有效黏附率分别为298.42±16.57和12.38±3.26 bubbles/per field，两者之间具有显著性差异(P < 0.01)。.③ 体内超声分子成像结果显示，在第7天、第12天和第17天，MMP-1靶向微泡的dTE信号强度显著高于对照组微泡(第7天，41.21±15.00 vs . 2.25±0.6 a.u, P < 0.05; 第12天，124.64±5.19 vs 11.13±1.13 a.u, P < 0.05; 第17天，(332.01±64.88 vs 42.99±11.9 a.u, P < 0.01)。并经免疫荧光检测分析证实MMP-1在喉鳞状细胞癌中的表达，且随着肿瘤的生长逐渐升高。.④ 常规超声、超声造影、增强CT对喉癌术前T分期的准确率差异无统计学意义 (P≥0.500)， 超声造影评价喉癌的喉软骨侵犯具有很高的敏感性及特异性，常规超声评价甲状软骨侵犯的敏感性、特异性、准确性分别为85.7%、75.0%、80.0%，超声造影评价甲状软骨受累的敏感性、特异性、准确性分别为92.9%、87.5%、90.0%，增强CT评价甲状软骨侵犯的敏感性、特异性、准确性分别为78.6%、87.5%、83.3% (P≥0.250)。.在成果上.1） 在本课题资助下，课题组共撰写科研论著5篇，其中已发表论文4篇，SCI收录4篇。.2） 培养硕士研究生3人。