CELL INTELLIGENCE · 肿瘤细胞治疗研究
肿瘤细胞治疗研究
英文原题：Immunohistochemical Detection of 5T4 in Renal Cell Carcinoma.
Immunohistochemical Detection of 5T4 in Renal Cell Carcinoma.
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5T4（由TPBG编码的滋养层糖蛋白）是一种癌/睾丸抗原，在肾细胞癌（RCC）及许多其他癌症中高表达，但在正常组织中很少表达。由于缺乏经过验证的5T4特异性免疫组织化学（IHC）试剂，将5T4开发为预后生物标志物以及通过新兴的受体工程化细胞免疫疗法直接靶向5T4的兴趣一直受到阻碍。
我们测试了4种市售单克隆抗体（mAb）用于检测福尔马林固定、石蜡包埋的RCC和正常组织中的5T4。使用亲本和TPBG编辑的A498细胞，3种mAb显示出5T4特异性。
进一步分析聚焦于2种染色最稳健的mAb（MBS1750093、Ab134162）。在包含263个肾脏肿瘤的组织微阵列上进行IHC显示，这2种mAb的染色一致性很高，从嫌色细胞RCC的0.80到晚期透明细胞RCC（ccRCC）的0.89不等。最敏感的MBS1750093在乳头状RCC（92.2%）>晚期ccRCC（60.0%）>嫌色细胞RCC（43.6%）>局限性ccRCC（39.6%）>嗜酸细胞瘤（22.7%）中表现出2+/3+染色。RNA原位杂交也显示高水平的TPBG RNA最常存在于乳头状和晚期ccRCC中。在晚期ccRCC中，有5T4表达较高与区域或远处转移的趋势。正常器官对照显示无染色或弱染色，例外的是IHC在肾小球和远端小管中显示局灶性中度染色。这些数据确定了适用于检测福尔马林固定、石蜡包埋组织中5T4的mAb，并证明了患者间和组织学亚型异质性。
我们经过验证的5T4 IHC方案将促进生物标志物研究并支持5T4的治疗性靶向。
5T4 (trophoblast glycoprotein encoded by TPBG ) is a cancer/testis antigen highly expressed in renal cell carcinoma (RCC) and many other cancers but rarely in normal tissues. Interest in developing 5T4 as a prognostic biomarker and direct targeting of 5T4 by emerging receptor-engineered cellular immunotherapies has been hampered by the lack of validated 5T4-specific reagents for immunohistochemistry (IHC).
We tested 4 commercially available monoclonal antibodies (mAbs) for the detection of 5T4 in formalin-fixed, paraffin-embedded RCC and normal tissues. Using parental and TPBG -edited A498 cells, 3 mAbs showed 5T4 specificity.
Further analyses focused on 2 mAbs with the most robust staining (MBS1750093, Ab134162). IHC on tissue microarrays incorporating 263 renal tumors showed high staining concordance of these 2 mAbs ranging from 0. 80 in chromophobe RCC to 0. 89 in advanced clear cell RCC (ccRCC). MBS1750093, the most sensitive, exhibited 2+/3+ staining in papillary RCC (92. 2%) > advanced ccRCC (60. 0%) > chromophobe RCC (43. 6%) > localized ccRCC (39. 6%) > oncocytoma (22. 7%).
RNA in situ hybridization also revealed high levels of TPBG RNA were present most frequently in papillary and advanced ccRCC. In advanced ccRCC, there was a trend towards higher 5T4 expression and regional or distant metastases. Normal organ controls showed no or weak staining with the exception of focal moderate staining in kidney glomeruli and distal tubules by IHC. These data identify mAbs suitable for detecting 5T4 in formalin-fixed, paraffin-embedded tissues and demonstrate both interpatient and histologic subtype heterogeneity.
Our validated 5T4 IHC protocol will facilitate biomarker studies and support the therapeutic targeting of 5T4.
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