National Institutes of Health, National Cancer Institute, Center for Cancer Research, Surgery Branch, Bethesda, Maryland, USA.
U.S. Food and Drug Administration, Center for Drug Evaluation and Research, Silver Spring, Maryland, USA.
Hum Gene Ther. 2021 Jul;32(13-14):730-743. doi: 10.1089/hum.2020.215. Epub 2021 Feb 22.
Chimeric antigen receptors (CARs) are artificial fusion proteins that incorporate antigen-recognition domains and T cell signaling domains. CD30 is a cell surface protein expressed on Hodgkin's lymphoma, some T cell lymphomas, and some B cell lymphomas. CD30 has a restricted expression pattern in normal cells, so CD30 has good potential as a clinical target for CAR T cells. We compared three different anti-CD30 CAR designs incorporating a single-chain variable fragment derived from the 5F11 fully human monoclonal antibody. 5F11-28Z has hinge, transmembrane, and costimulatory domains from CD28 and a CD3ζ T cell activation domain. 5F11-CD828Z has hinge and transmembrane domains from CD8α, a CD28 costimulatory domain, and a CD3ζ T cell activation domain. 5F11-CD8BBZ is identical to 5F11-CD828Z, except for the replacement of the CD28 moiety with a 4-1BB moiety. We found that T cells expressing 5F11-CD8BBZ had lower levels of CD30-specific degranulation and cytokine release compared with CD28-containing CARs. When compared to the CD28-containing CARs, T cells expressing 5F11-CD8BBZ had higher levels of nonspecific functional activity, including degranulation, cytokine release, and proliferation, when stimulated with CD30-negative target cells. We established tumors in nod-scid common gamma-chain deficient (NSG) mice and treated the tumors with T cells expressing different CARs. T cells expressing 5F11-28Z were most effective at eradicating tumors. T cells expressing 5F11-CD828Z had intermediate effectiveness, and T cells expressing 5F11-CD8BBZ were least effective. CD30 T cells are lost from cultures of T cells containing 5F11-28Z-expressing T cells. This indicated the killing of CD30 T cells by the 5F11-28Z-expressing T cells. Despite this, the number of T cells in the cultures consistently accumulated to numbers needed for use in a clinical trial. Based on all and murine experiments comparing the different CARs, we selected 5F11-28Z for further development, and we have initiated a clinical trial testing 5F11-28Z T cells.
嵌合抗原受体 (CAR) 是一种人工融合蛋白，它包含抗原识别结构域和 T 细胞信号结构域。CD30 是一种在霍奇金淋巴瘤、一些 T 细胞淋巴瘤和一些 B 细胞淋巴瘤上表达的细胞表面蛋白。CD30 在正常细胞中的表达模式受限，因此 CD30 作为 CAR T 细胞的临床靶点具有很好的潜力。我们比较了三种不同的抗 CD30 CAR 设计，它们都包含来自 5F11 全人源单克隆抗体的单链可变片段。5F11-28Z 具有来自 CD28 的铰链、跨膜和共刺激结构域以及 CD3ζ T 细胞激活结构域。5F11-CD828Z 具有来自 CD8α 的铰链和跨膜结构域、CD28 共刺激结构域和 CD3ζ T 细胞激活结构域。5F11-CD8BBZ 与 5F11-CD828Z 相同，只是用 4-1BB 部分取代了 CD28 部分。我们发现表达 5F11-CD8BBZ 的 T 细胞与包含 CD28 的 CAR 相比，CD30 特异性脱颗粒和细胞因子释放水平较低。与包含 CD28 的 CAR 相比，当用 CD30 阴性靶细胞刺激时，表达 5F11-CD8BBZ 的 T 细胞具有更高水平的非特异性功能活性，包括脱颗粒、细胞因子释放和增殖。我们在 nod-scid 共同 γ 链缺陷 (NSG) 小鼠中建立肿瘤，并使用表达不同 CAR 的 T 细胞治疗肿瘤。表达 5F11-28Z 的 T 细胞最有效地根除肿瘤。表达 5F11-CD828Z 的 T 细胞具有中等疗效，而表达 5F11-CD8BBZ 的 T 细胞疗效最差。表达 5F11-28Z 的 T 细胞中的 CD30 T 细胞从 T 细胞培养物中丢失。这表明表达 5F11-28Z 的 T 细胞杀死了 CD30 T 细胞。尽管如此，培养物中的 T 细胞数量仍不断积累到临床试验所需的数量。基于所有人和鼠实验比较不同的 CAR，我们选择 5F11-28Z 进行进一步开发，并已启动一项临床试验，以测试 5F11-28Z T 细胞。
