Duke Brain Tumor Immunotherapy Program, Department of Neurosurgery, Duke University Medical Center, Durham, NC, United States of America.
Department of Biomedical Engineering, Duke University, Durham, NC, United States of America.
PLoS One. 2020 Jul 31;15(7):e0236374. doi: 10.1371/journal.pone.0236374. eCollection 2020.
We recently reported the development of a fully-human, CD3-binding bispecific antibody for immunotherapy of malignant glioma. To translate this therapeutic (hEGFRvIII-CD3- bi-scFv) to clinical trials and to help further the translation of other similar CD3-binding therapeutics, some of which are associated with neurologic toxicities, we performed a good laboratory practice (GLP) toxicity study to assess for potential behavioral, chemical, hematologic, and pathologic toxicities including evaluation for experimental autoimmune encephalomyelitis (EAE). To perform this study, male and female C57/BL6 mice heterozygous for the human CD3 transgene (20/sex) were allocated to one of four designated groups. All animals were administered one dose level of hEGFRvIII-CD3 bi-scFv or vehicle control. Test groups were monitored for feed consumption, changes in body weight, and behavioral disturbances including signs of EAE. Urinalysis, hematologic, and clinical chemistry analysis were also performed. Vehicle and test chemical-treated groups were humanely euthanized 48 hours or 14 days following dose administration. Complete gross necropsy of all tissues was performed, and selected tissues plus all observed gross lesions were collected and evaluated for microscopic changes. This included hematoxylin-eosin histopathological evaluation and Fe-ECR staining for myelin sheath enumeration. There were no abnormal clinical observations or signs of EAE noted during the study. There were no statistical changes in food consumption, body weight gain, or final body weight among groups exposed to hEGFRvIII-CD3 bi-scFv compared to the control groups for the 2- and 14-day timepoints. There were statistical differences in some clinical chemistry, hematologic and urinalysis endpoints, primarily in the females at the 14-day timepoint (hematocrit, calcium, phosphorous, and total protein). No pathological findings related to hEGFRvIII-CD3 bi-scFv administration were observed. A number of gross and microscopic observations were noted but all were considered to be incidental background findings. The results of this study allow for further translation of this and other important CD3 modulating bispecific antibodies.
我们最近报道了一种完全人源化的、针对 CD3 的双特异性抗体的开发，用于恶性神经胶质瘤的免疫治疗。为了将这种治疗性（hEGFRvIII-CD3-双 scFv）转化为临床试验，并帮助进一步转化其他类似的与神经毒性相关的 CD3 结合治疗剂，我们进行了一项良好实验室规范（GLP）毒性研究，以评估潜在的行为、化学、血液学和病理学毒性，包括实验性自身免疫性脑脊髓炎（EAE）的评估。为了进行这项研究，我们将携带人类 CD3 转基因的雄性和雌性 C57/BL6 小鼠（每性别 20 只）分配到四个指定组之一。所有动物都接受了一个剂量水平的 hEGFRvIII-CD3 双 scFv 或载体对照。测试组监测饲料消耗、体重变化以及行为障碍，包括 EAE 的迹象。还进行了尿液分析、血液学和临床化学分析。在给药后 48 小时或 14 天，对载体和测试化学处理组的动物进行安乐死。对所有组织进行完整的大体解剖，并采集和评估选定的组织和所有观察到的大体病变，以评估微观变化。这包括苏木精-伊红组织病理学评估和铁-ECR 染色以计数髓鞘。在研究期间，没有观察到异常的临床观察或 EAE 迹象。与对照组相比，暴露于 hEGFRvIII-CD3 双 scFv 的各组在 2 天和 14 天时间点的食物消耗、体重增加或最终体重没有统计学上的变化。在一些临床化学、血液学和尿液分析终点，主要是在 14 天时间点的雌性中，存在统计学差异（红细胞压积、钙、磷和总蛋白）。未观察到与 hEGFRvIII-CD3 双 scFv 给药相关的病理发现。注意到许多大体和显微镜下的观察结果，但都被认为是偶然的背景发现。这项研究的结果允许进一步转化这种和其他重要的 CD3 调节双特异性抗体。
