Development of a high affinity Anticalin® directed against human CD98hc for theranostic applications
Development of a high affinity Anticalin® directed against human CD98hc for theranostic applications
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DOI：
10.7150/thno.38968
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发表时间：
2020-01-01
期刊：
影响因子：
12.4
通讯作者：
Skerra, Arne
中科院分区：
文献类型：
--
作者：
Deuschle, Friedrich-Christian;Morath, Volker;Skerra, Arne
增强的氨基酸供应和失调的整合素信号是癌症的两个标志，是细胞转移转化的关键。由于其在这两个过程的交叉点上的功能，CD98hc在临床上被观察到在各种恶性肿瘤中过表达，从而使其成为一个有希望的肿瘤靶点。方法：利用定向进化和蛋白设计技术，以脂质钙素2 （Lcn2）为支架，构建抗人CD98hc外结构域（hCD98hcED）的抗血清素蛋白。x射线结构分析鉴定了antialin候选先导分子识别的表位。使用PASylation (R)技术调节血浆半衰期的antialin用Zr-89标记，并通过cd98阳性肿瘤移植小鼠的正电子发射断层扫描（PET）进行研究。结果：antialin P3D11以皮摩尔亲和力结合CD98hc，并在hCD98hcED的溶剂暴露膜远端识别出由几个糖基化位点包围的突出环结构。体外研究显示antialin对多种表达cd98hc的人肿瘤细胞系具有特异性结合活性，提示其在癌症研究中具有更广泛的适用性。使用优化的zr -89标记的antialin对携带人类前列腺癌异种移植的小鼠进行PET/CT成像，显示出强大的特异性示踪剂积累（8.6 +/- 1.1% ID/g），以及良好的肿瘤与血液比为11.8。结论：我们的发现为利用CD98hc进行非侵入性生物医学成像提供了第一个概念证明。新的基于抗钙素的α - hCD98hc放射性药物构成了肿瘤临床前和潜在临床应用的有前途的工具。
Enhanced amino acid supply and dysregulated integrin signaling constitute two hallmarks of cancer and are pivotal for metastatic transformation of cells. In line with its function at the crossroads of both processes, overexpression of CD98hc is clinically observed in various cancer malignancies, thus rendering it a promising tumor target.Methods: We describe the development of Anticalin proteins based on the lipocalin 2 (Lcn2) scaffold against the human CD98hc ectodomain (hCD98hcED) using directed evolution and protein design. X-ray structural analysis was performed to identify the epitope recognized by the lead Anticalin candidate. The Anticalin - with a tuned plasma half-life using PASylation (R) technology - was labeled with Zr-89 and investigated by positron emission tomography (PET) of CD98-positive tumor xenograft mice.Results: The Anticalin P3D11 binds CD98hc with picomolar affinity and recognizes a protruding loop structure surrounded by several glycosylation sites within the solvent exposed membrane-distal part of the hCD98hcED. In vitro studies revealed specific binding activity of the Anticalin towards various CD98hc-expressing human tumor cell lines, suggesting broader applicability in cancer research. PET/CT imaging of mice bearing human prostate carcinoma xenografts using the optimized and Zr-89-labeled Anticalin demonstrated strong and specific tracer accumulation (8.6 +/- 1.1 %ID/g) as well as a favorable tumor-to-blood ratio of 11.8.Conclusion: Our findings provide a first proof of concept to exploit CD98hc for non-invasive biomedical imaging. The novel Anticalin-based alpha hCD98hc radiopharmaceutical constitutes a promising tool for preclinical and, potentially, clinical applications in oncology.