VHH-Photosensitizer Conjugates for Targeted Photodynamic Therapy of Met-Overexpressing Tumor Cells
VHH-Photosensitizer Conjugates for Targeted Photodynamic Therapy of Met-Overexpressing Tumor Cells
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DOI：
10.3390/antib8020026
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发表时间：
2019-06-01
期刊：
影响因子：
4.7
通讯作者：
Oliveira, Sabrina
中科院分区：
文献类型：
--
作者：
Heukers, Raimond;Mashayekhi, Vida;Oliveira, Sabrina
光动力疗法（PDT）是通过在光敏剂（PS）的局部照射时局部产生毒性活性氧来杀死（癌）细胞的方法。通过开发新的水溶性PS和通过与肿瘤靶向抗体缀合来特异性递送PS，进一步增强了PDT的特异性。为了改善组织渗透和缩短光敏性，我们最近引入了纳米抗体，也称为VHH（来自美洲驼重链抗体重链的可变结构域），用于过表达表皮生长因子受体（EGFR）的癌细胞的靶向PDT。另一种癌症相关受体肝细胞生长因子受体（HGFR、c-Met或Met）的过表达和活化也参与多种恶性肿瘤的进展和转移。在这项研究中，我们评估是否抗Met VHH共轭PS也可以作为一种生物药物靶向PDT。向靶向Met的SEMA（脑信号蛋白样）亚结构域的VHH提供C-末端标签，其允许从酵母上清液直接纯化和使用马来酰亚胺化学定向缀合至PS IRDye 700DX。产生的抗Met VHH-PS显示出纳摩尔结合亲和力，并且在照射后，以纳摩尔效力特异性杀死MKN 45细胞。这项研究表明，Met也可以作为靶向PDT的膜靶点。
Photodynamic therapy (PDT) is an approach that kills (cancer) cells by the local production of toxic reactive oxygen species upon the local illumination of a photosensitizer (PS). The specificity of PDT has been further enhanced by the development of a new water-soluble PS and by the specific delivery of PS via conjugation to tumor-targeting antibodies. To improve tissue penetration and shorten photosensitivity, we have recently introduced nanobodies, also known as VHH (variable domains from the heavy chain of llama heavy chain antibodies), for targeted PDT of cancer cells overexpressing the epidermal growth factor receptor (EGFR). Overexpression and activation of another cancer-related receptor, the hepatocyte growth factor receptor (HGFR, c-Met or Met) is also involved in the progression and metastasis of a large variety of malignancies. In this study we evaluate whether anti-Met VHHs conjugated to PS can also serve as a biopharmaceutical for targeted PDT. VHHs targeting the SEMA (semaphorin-like) subdomain of Met were provided with a C-terminal tag that allowed both straightforward purification from yeast supernatant and directional conjugation to the PS IRDye700DX using maleimide chemistry. The generated anti-Met VHH-PS showed nanomolar binding affinity and, upon illumination, specifically killed MKN45 cells with nanomolar potency. This study shows that Met can also serve as a membrane target for targeted PDT.