Establishment of an antibody specific for AMIGO2 improves immunohistochemical evaluation of liver metastases and clinical outcomes in patients with colorectal cancer.
Establishment of an antibody specific for AMIGO2 improves immunohistochemical evaluation of liver metastases and clinical outcomes in patients with colorectal cancer.
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DOI：
10.1186/s13000-021-01176-2
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发表时间：
2022-01-30
影响因子：
2.6
通讯作者：
Okada F
中科院分区：
文献类型：
--
作者：
Goto K;Osaki M;Izutsu R;Tanaka H;Sasaki R;Tanio A;Satofuka H;Kazuki Y;Yamamoto M;Kugoh H;Ito H;Oshimura M;Fujiwara Y;Okada F
人两性蛋白诱导基因和开放阅读框（AMIGO）被鉴定为一种新型的I型跨膜蛋白细胞粘附分子。 AMIGO2是AMIGO家族的三个成员（AMIGO1、2和3）之一，它们之间在氨基酸水平上的相似性约为40%。我们之前已经证明 AMIGO2 是肝转移的驱动因素。使用市售的抗 AMIGO2 小鼠单克隆抗体克隆 sc-373699 (sc mAb) 对结直肠癌 (CRC) 中的 AMIGO2 表达进行免疫组织化学分析，与肝转移和不良预后相关。然而，sc mAb 被发现与 AMIGO 家族中的所有三种分子发生交叉反应。我们生成了针对人 AMIGO2 的大鼠单克隆抗体克隆 rTNK1A0012 (rTNK mAb)。 rTNK mAb 使用先前使用 sc mAb 报告的相同 CRC 组织样本重新评估 AMIGO2 表达与肝转移/临床结果之间的关联。蛋白质印迹分析显示，rTNK mAb 被鉴定为对 AMIGO2 蛋白具有特异性，并且不与 AMIGO1 和 AMIGO3 发生交叉反应。 rTNK mAb 和 sc mAb 显示较高的 AMIGO2 表达，这与肝转移的高频率相关（分别为 65.3% 和 47.5%），而多变量分析显示 AMIGO2 表达是肝转移的独立预后因素（p = 7.930E-10 和 p = 1.707E-5）。 Kaplan-Meier 分析显示，rTNK mAb (p = 0.004)，而非 sc mAb (p = 0.107)，预测 AMIGO2 高表达患者的总生存期较差。与 sc mAb (p = 0.001) 相比，rTNK mAb (p = 0.00004) 的 AMIGO2 表达与较差的疾病特异性生存率之间的关系具有更高的显着性。这些结果表明，开发的rTNK1A0012 mAb是一种通过免疫组织化学特异性识别AMIGO2的抗体，可以成为诊断检测高AMIGO2表达CRC患者肝转移和较差预后的更可靠和适用的方法。在线版本包含可在 10.1186/s13000-021-01176-2 获取的补充材料。
The human amphoterin-induced gene and open reading frame (AMIGO) was identified as a novel cell adhesion molecule of type I transmembrane protein. AMIGO2 is one of three members of the AMIGO family (AMIGO1, 2, and 3), and the similarity between them is approximately 40% at the amino acid level. We have previously shown that AMIGO2 functions as a driver of liver metastasis. Immunohistochemical analysis of AMIGO2 expression in colorectal cancer (CRC) using a commercially available anti-AMIGO2 mouse monoclonal antibody clone sc-373699 (sc mAb) correlated with liver metastasis and poor prognosis. However, the sc mAb was found to be cross-reactive with all three molecules in the AMIGO family. We generated a rat monoclonal antibody clone rTNK1A0012 (rTNK mAb) for human AMIGO2. The rTNK mAb was used to re-evaluate the association between AMIGO2 expression and liver metastases/clinical outcomes using the same CRC tissue samples previously reported with sc mAb. Western blot analysis revealed that a rTNK mAb was identified as being specific for AMIGO2 protein and did not cross-react with AMIGO1 and AMIGO3. The rTNK mAb and sc mAb showed higher AMIGO2 expression, which correlates with a high frequency of liver metastases (65.3% and 47.5%, respectively), while multivariate analysis showed that AMIGO2 expression was an independent prognostic factor for liver metastases (p = 7.930E-10 and p = 1.707E-5). The Kaplan-Meier analyses showed that the rTNK mAb (p = 0.004), but not sc mAb (p = 0.107), predicted worse overall survival in patients with high AMIGO2 expression. The relationship between AMIGO2 expression and poor disease-specific survival showed a higher level of significance for rTNK mAb (p = 0.00004) compared to sc mAb (p = 0.001). These results indicate that the developed rTNK1A0012 mAb is an antibody that specifically recognizes AMIGO2 by immunohistochemistry and can be a more reliable and applicable method for the diagnostic detection of liver metastases and worse prognosis in patients with high AMIGO2-expressing CRC. The online version contains supplementary material available at 10.1186/s13000-021-01176-2.