PO.TB01.01 · 肿瘤生物学
一种靶向血管生成的mRNA疫苗在两种小鼠黑色素瘤模型中引发强健的免疫应答并抑制肿瘤生长和血管生成
An mRNA vaccine targeting angiogenesis elicits a robust immune response and inhibits tumor growth and angiogenesis in two mouse melanoma models
作者与单位 Authors & Affiliations
摘要 Abstract
中文摘要
背景：血管生成对肿瘤生长、转移和免疫抑制微环境的建立至关重要。通过支持营养输送和调节免疫细胞浸润，肿瘤驱动的血管生成促进恶性进展和免疫逃逸。在其关键调节因子中，肾上腺髓质素（AM）是一种在实体瘤中经常过表达的强效促血管生成因子。为探索其治疗潜力，我们开发了一种mRNA疫苗，编码由小的钥孔血蓝蛋白（KLH）肽（作为半抗原）连接到小鼠AM组成的融合抗原。方法：体外转录的mRNA被封装在脂质纳米颗粒（LNPs）中，并施用于C57BL/6小鼠，以空LNPs作为对照。该疫苗在两种不同的黑色素瘤模型中进行评估：由静脉注射B16-F10黑色素瘤细胞诱导的实验性肺转移模型，以及皮下黑色素瘤模型。经过四次免疫后，注射黑色素瘤细胞，随后进行第五次免疫。一旦肿瘤达到人道终点即处死小鼠，并分析血液和器官。结果：与对照相比，接种疫苗的小鼠表现出抗AM IgG滴度（p= 0.040）和CD8+ T细胞数量（p=0.033）的显著增加。在肺转移模型中，免疫导致肺转移灶的数量（p=0.028）和大小（p=0.020）均显著减少，以及肿瘤相关血管数量的减少（p=0.046），且如在SMAA-GFP转基因小鼠中评估的，未破坏正常血管系统。在皮下模型中，接种疫苗延迟了肿瘤起始（p=0.005）并减小了肿瘤体积（p=0.004），伴随肿瘤血管生成的减少（p=0.045）。重要的是，未观察到全身毒性或体重的显著变化。结论：通过靶向血管生成，KLH-AM mRNA疫苗有效地削弱了黑色素瘤模型中的肿瘤生长。这些结果凸显了AM作为驱动肿瘤血管化的重要促血管生成因子，并支持进一步开发AM导向的mRNA治疗性疫苗作为一种破坏血管生成依赖性肿瘤进展的策略。本研究由西班牙国家研究局（AEI）项目PID2022-136735OB-I00资助。
查看英文原文 English abstract
Background: Angiogenesis is essential for tumor growth, metastasis, and the establishment of an immunosuppressive microenvironment. By supporting nutrient delivery and modulating immune cell infiltration, tumor-driven angiogenesis promotes both malignant progression and immune evasion. Among its key regulators, adrenomedullin (AM) is a potent pro-angiogenic factor frequently overexpressed in solid tumors. To explore its therapeutic potential, we developed an mRNA vaccine encoding a fusion antigen composed of a small keyhole limpet hemocyanin (KLH) peptide, as a hapten, linked to mouse AM.
Methods: The in vitro transcribed mRNA was encapsulated in lipid nanoparticles (LNPs) and administered to C57BL/6 mice, with empty LNPs serving as controls. The vaccine was evaluated in two distinct melanoma models: an experimental lung metastasis model induced by intravenous injection of B16-F10 melanoma cells, and a subcutaneous melanoma model. After four immunizations, melanoma cells were injected, followed by a fifth immunization. Mice were sacrificed once tumors reached humane endpoints and blood and organs were analyzed.
Results: Vaccinated mice exhibited significant increases in anti-AM IgG titers (p= 0.040) and CD8+ T cell numbers (p=0.033) compared to controls. In the lung metastasis model, immunization resulted in a significant reduction in both the number (p=0.028) and size of lung metastases (p=0.020) as well as a decrease in the number of tumor-associated blood vessels (p=0.046), without disrupting normal vasculature as assessed in SMAA-GFP transgenic mice. In the subcutaneous model, vaccination delayed tumor initiation (p=0.005) and reduced tumor volume (p=0.004), accompanied by decreased tumor angiogenesis (p=0.045). Importantly, no systemic toxicity or significant changes in weight were observed.
Conclusion: By targeting angiogenesis, the KLH-AM mRNA vaccine effectively impaired tumor growth in melanoma models. These results highlight AM as an important pro-angiogenic factor driving tumor vascularization and support further development of AM-directed mRNA therapeutic vaccination as a strategy to disrupt angiogenesis-dependent tumor progression.This study was funded by Agencia Estatal de Investigación (AEI, Spain)'s project PID2022-136735OB-I00.
利益披露 Disclosure
S. Tadic, None..
L. Ochoa-Callejero, None..
J. Narro Íñiguez, None..
J. García-Sanmartín, None..
A. Martínez, None.