Department of Pathology and Translational Research, Gifu University Graduate School of Medicine, Gifu, Japan.
Department of Pathology, Matsunami General Hospital, Gifu, Japan.
Thorac Cancer. 2022 Sep;13(18):2566-2573. doi: 10.1111/1759-7714.14591. Epub 2022 Aug 2.
As mesothelioma generally has an unfavorable prognosis, further advances are needed to improve the outcomes in patients with mesothelioma. In the present study, we generated and characterized a monoclonal antibody that could inhibit mesothelioma cell proliferation in a xenotransplantation mouse model.
We generated monoclonal antibodies by immunizing mice with cultured mesothelioma cells. These antibodies were then characterized by immunofluorescence staining, immunohistochemical staining, secondary antibody-drug conjugate assay, antibody inoculation in a xenotransplantation mesothelioma mouse model, and mass spectrometry followed by small interfering RNA (siRNA) analysis. 5' rapid amplification of complementary DNA ends followed by sequencing was performed to deduce the amino acid sequences of the variable regions of the light and heavy chains of AX10.
An IgG2b κ-type AX10 antibody against the cell surface membrane of sarcomatoid mesothelioma cells was generated. AX10 immunoreactivity was detected in 12 out of 22 different mesothelioma tissue specimens, but there was little AX10 immunoreactivity in a normal human tissue array. AX10 decreased Matrigel invasion by MPM-1 cells but did not affect cell proliferation. Notably, AX10 significantly inhibited the proliferation of MPM-1 cells xenotransplanted into Severe combined immunodeficiency-Nonobese diabetic mice. Matrix-assisted laser desorption ionization time-of-flight mass spectrometry followed by siRNA silencing indicated that AX10 reacted to a unique alternatively spliced isoform of sarcolemma-associated protein. AX10 is composed of as yet unregistered amino acid sequences in its variable region.
AX10 could have therapeutic potential for patients with sarcomatoid mesothelioma.
由于间皮瘤的预后通常较差,因此需要进一步的进展来改善间皮瘤患者的治疗效果。在本研究中,我们生成并鉴定了一种单克隆抗体,该抗体可在异种移植小鼠模型中抑制间皮瘤细胞的增殖。
我们通过用培养的间皮瘤细胞免疫小鼠来生成单克隆抗体。然后通过免疫荧光染色、免疫组织化学染色、第二抗体-药物偶联物测定、在异种移植间皮瘤小鼠模型中接种抗体以及质谱分析结合小干扰 RNA(siRNA)分析对这些抗体进行了鉴定。通过 5' 快速扩增互补 DNA 末端测序来推断 AX10 的轻链和重链可变区的氨基酸序列。
生成了一种针对肉瘤样间皮瘤细胞膜的 IgG2b κ 型 AX10 抗体。AX10 免疫反应性在 22 个不同的间皮瘤组织标本中的 12 个中被检测到,但在正常人类组织阵列中几乎没有 AX10 免疫反应性。AX10 可减少 MPM-1 细胞在 Matrigel 中的侵袭,但不影响细胞增殖。值得注意的是,AX10 显著抑制了异种移植到严重联合免疫缺陷-非肥胖型糖尿病小鼠中的 MPM-1 细胞的增殖。基质辅助激光解吸电离飞行时间质谱分析结合 siRNA 沉默表明,AX10 与细胞膜相关蛋白的独特剪接异构体反应。AX10 在其可变区由尚未注册的氨基酸序列组成。
AX10 可能对肉瘤样间皮瘤患者具有治疗潜力。