PURPOSE Although recent advances in molecular target therapy have improved the survival of breast cancer patients, high cost and frequent hospital visits result in both societal and individual burden. To reduce these problems, it has been proposed to produce antibodies in vivo. Here, we constructed gene-transduced human ceiling culture-derived proliferative adipocytes secreting anti-HER2 antibody (HER2-ccdPAs) and evaluated their ability to secrete antibody and mediate an anti-tumor effect. METHODS Plasmid lentivirus was used as a recipient for anti-HER2 antibody cDNA and transduced into human proliferative adipocyte. Secretory antibody expression was evaluated by ELISA and western blot. Specific binding of secretory antibody to HER2 was examined by immunofluorescence analysis. Direct and indirect anti-tumor effects of supernatants from HER2-ccdPAs were evaluated using BT474 (HER2+) and MDA-MB-231 (HER2-) breast cancer cell lines. Additionally, whether adipocyte differentiation affects antibody secretion was investigated using supernatant collected from different cell maturation states. RESULTS Anti-HER2 antibody was identified in the supernatant from HER2-ccdPAs and its production increased with the differentiation into mature adipocyte. Antibodies in supernatants from HER2-ccdPAs bound to HER2-positive breast cancer cells similar to trastuzumab. Supernatant from HER2-ccdPAs inhibited the proliferation of BT474 but not MDA-MB-231 cells, and downregulated AKT phosphorylation in BT474 cells compared with controls. Supernatants from HER2-ccdPAs also had an indirect anti-tumor effect on BT474 cells through ADCC. Additionally, Single inoculation of HER2-ccdPAs showed an anti-tumor effect in BT474 xenograft model. CONCLUSIONS HER2-ccdPAs might be useful for cell-based gene therapy. This system could be a platform for various antibody therapies.

中文翻译：

---

使用基因转导的脂肪细胞对HER2阳性乳腺癌进行抗HER2抗体治疗。

目的尽管分子靶向治疗的最新进展提高了乳腺癌患者的生存率，但高昂的费用和频繁的医院就诊带来了社会和个人负担。为了减少这些问题，已经提出在体内产生抗体。在这里，我们构建了分泌抗HER2抗体（HER2-ccdPAs）的基因转导的人类天花板培养衍生的增殖性脂肪细胞，并评估了它们分泌抗体和介导抗肿瘤作用的能力。方法采用质粒慢病毒作为抗HER2抗体cDNA的受体，并转导至人增殖性脂肪细胞中。通过ELISA和蛋白质印迹评估分泌抗体的表达。通过免疫荧光分析检查了分泌抗体与HER2的特异性结合。使用BT474（HER2 +）和MDA-MB-231（HER2-）乳腺癌细胞系评估了HER2-ccdPAs上清液的直接和间接抗肿瘤作用。另外，使用从不同细胞成熟状态收集的上清液研究了脂肪细胞的分化是否影响抗体的分泌。结果在HER2-ccdPAs的上清液中鉴定出抗HER2抗体，并随着向成熟脂肪细胞的分化而增加其产量。与曲妥珠单抗类似，HER2-ccdPAs上清液中的抗体与HER2阳性乳腺癌细胞结合。与对照组相比，HER2-ccdPAs的上清液抑制BT474的增殖，但抑制MDA-MB-231细胞的增殖，并抑制BT474细胞的AKT磷酸化。HER2-ccdPAs的上清液也通过ADCC对BT474细胞具有间接的抗肿瘤作用。此外，单次接种HER2-ccdPAs在BT474异种移植模型中显示出抗肿瘤作用。结论HER2-ccdPAs可能对基于细胞的基因治疗有用。该系统可以是各种抗体疗法的平台。