Novel EBV LMP1 C-terminal domain binding affibody molecules as potential agents for in vivo molecular imaging diagnosis of nasopharyngeal carcinoma
Novel EBV LMP1 C-terminal domain binding affibody molecules as potential agents for in vivo molecular imaging diagnosis of nasopharyngeal carcinoma
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DOI：
10.1007/s00253-021-11559-6
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发表时间：
2021-09-10
影响因子：
5
通讯作者：
Zhang, Lifang
中科院分区：
文献类型：
--
作者：
Kamara, Saidu;Guo, Yanru;Zhang, Lifang
鼻咽癌（NPC）始终与EB病毒（EBV）潜伏感染相关，在中国南方和东南亚很常见。病毒潜伏膜蛋白LMP1和LMP2在鼻咽癌组织中持续表达； LMP1（LMP1 C 端）和 LMP2A（LMP2A N 端）蛋白的胞质结构域对于维持潜伏期至关重要，并且可以改变宿主细胞信号传导以促进肿瘤生长和进展。因此，靶向LMP1或LMP2癌蛋白对于鼻咽癌的诊断和靶向治疗越来越感兴趣。 Affibody 分子是一类新型的小亲和力工程支架蛋白，已显示出在治疗、诊断和生物技术应用方面的巨大潜力。最近，放射性标记的 HER2 特异性亲和体分子已被证明可用于 HER2 表达肿瘤的成像。在本研究中，我们报告了通过生物淘选从随机肽展示噬菌体库中选择的三种新型 EBV LMP1 C 末端 (EBV LMP1-C) 结构域亲和体分子（Z(LMP1-C)15、Z(LMP1-C)114 和 Z(LMP1-C)277），并用于荷瘤裸鼠的分子成像。表面等离子共振 (SPR)、间接免疫荧光和免疫组织化学 (IHC) 清楚地表明，所有三种选定的 Affibody 分子在与 EBV LMP1 蛋白结合方面均具有高亲和力和特异性。此外，体内肿瘤成像显示，Dylight-755标记的亲和体分子注射后（1小时）在肿瘤部位迅速积累，然后在EBV阳性NPC异种移植小鼠模型中持续维持24小时。总之，我们的研究结果强调了 Z(LMP1-C) 亲和体分子作为 EBV 相关 NPC 肿瘤特异性分子成像剂的潜在用途。
Nasopharyngeal carcinoma (NPC) is consistently associated with Epstein-Barr virus (EBV) latent infection and is common in Southern China and Southeast Asia. The viral latent membrane proteins LMP1 and LMP2 are persistently expressed in NPC tissues; the cytoplasmic domain of LMP1 (LMP1 C-terminal) and LMP2A (LMP2A N-terminal) proteins is essential for maintenance of latency and can alter host cell signaling to facilitate tumor growth and progression. Thus, targeting LMP1 or LMP2 oncoprotein has been an increasing interest for diagnosis and targeted therapy of NPC. Affibody molecules, a new class of small-affinity engineered scaffold proteins, have demonstrated high potential for therapeutics, diagnostics, and biotechnological applications. More recently, radiolabelled HER2-specific affibody molecules have demonstrated to be useful in imaging of HER2 expressing tumor. In this study, we report three novel EBV LMP1 C-terminal (EBV LMP1-C) domain affibody molecules (Z(LMP1-C)15, Z(LMP1-C)114, and Z(LMP1-C)277) were selected by biopanning from a random-peptide displayed phage library and used for molecular imaging in tumor-bearing nude mice. Surface plasmon resonance (SPR), indirect immunofluorescence, and immunohistochemistry (IHC) clearly showed that all three selected affibody molecules have high affinity and specificity in binding to EBV LMP1 protein. Moreover, in vivo tumor imaging revealed that Dylight-755-labeled affibody molecules accumulated rapidly in tumor site after injection (1 h) and then were continuously maintained for 24 h in EBV-positive NPC xenograft mice model. In conclusion, our findings highlight the potential use of Z(LMP1-C) affibody molecules as tumor-specific molecular imaging agents of EBV-associated NPC.