Development of novel monoclonal antibodies against CD109 overexpressed in human pancreatic cancer.
Development of novel monoclonal antibodies against CD109 overexpressed in human pancreatic cancer.
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DOI：
10.18632/oncotarget.25017
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发表时间：
2018-04-13
期刊：
影响因子：
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通讯作者：
Modjtahedi H
中科院分区：
文献类型：
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作者：
Arias-Pinilla GA;Dalgleish AG;Mudan S;Bagwan I;Walker AJ;Modjtahedi H
胰腺癌是最具侵袭性和致命性的癌症之一，迫切需要更有效的治疗药物。过表达的细胞表面抗原是以单抗为基础的药物治疗的理想靶点，但还没有一种药物被批准用于胰腺癌的治疗。在这里，我们报道了两种新的抗人胰腺癌细胞株BxPC-3的鼠单抗KU42.33C和KU43.13A的研制。通过采用ELISA法、流式细胞术、竞争分析和免疫沉淀-质谱仪等方法，我们发现这两株单抗针对CD109外区的两个不同的表位，这两个表位在人胰腺癌细胞系中有不同程度的过表达。单独使用这两种裸露抗体治疗并不影响肿瘤细胞的生长或体外迁移。在两株单抗中，只有KU42.33C可用于免疫印迹和免疫组织化学检测肿瘤细胞中CD109的表达。有趣的是，用单抗KU42.33C对人胰腺癌组织芯片进行免疫组织化学染色，发现65例人胰腺癌组织中有94%的CD109表达阳性，正常胰腺组织中无表达。我们的结果表明，这两个新的单抗是检测CD109在多种癌症，特别是胰腺癌患者的肿瘤标本和血清中表达水平的良好工具，并可用于研究其诊断、预后和预测价值。进一步的研究是有必要的，应该旨在揭示人源化形式或结合形式的这种抗体在肿瘤过度表达CD109抗原的患者中的治疗潜力。
Pancreatic cancer is one of the most aggressive and lethal types of cancer, and more effective therapeutic agents are urgently needed. Overexpressed cell surface antigens are ideal targets for therapy with monoclonal antibody (mAb)-based drugs, but none have been approved for the treatment of pancreatic cancer. Here, we report development of two novel mouse mAbs, KU42.33C and KU43.13A, against the human pancreatic cancer cell line BxPC-3. Using ELISA, flow cytometry, competitive assay and immunoprecipitation followed by mass spectrometry, we discovered that these two mAbs target two distinct epitopes on the external domain of CD109 that are overexpressed by varying amounts in human pancreatic cancer cell lines. Treatment with these two naked antibodies alone did not affect tumour cell growth or migration in vitro. Of the two mAbs, only KU42.33C was useful in determining the expression of CD109 in tumour cells by Western blot and immunohistochemistry. Interestingly, immunohistochemistry of human pancreatic carcinoma tissue arrays with mAb KU42.33C showed that 94% of the 65 human pancreatic adenocarcinoma cases were CD109 positive, with no expression in normal pancreatic tissues. Our results suggest that these two novel mAbs are excellent tools for determining the expression level of CD109 in the tumour specimens and sera of patients with a wide range of cancers, in particular pancreatic cancer, and for investigating its diagnostic, prognostic and predictive value. Further research is warranted and should aim to unravel the therapeutic potential of the humanised forms or conjugated versions of such antibodies in patients whose tumours overexpress CD109 antigen.