文章：

IL-15超激动剂的共表达促进GD2靶向CAR-NK细胞自我富集并在无IL-2条件下介导高效细胞杀伤

Co-Expression of an IL-15 Superagonist Facilitates Self-Enrichment of GD2-Targeted CAR-NK Cells and Mediates Potent Cell Killing in the Absence of IL-2

原文发布日期：29 August 2023

DOI: 10.3390/cancers15174310

类型: Article

开放获取: 是

英文摘要：

In contrast to T lymphocytes, natural killer (NK) cells do not require prior sensitization but are rapidly activated upon encountering virally infected or neoplastic cells. In addition, NK cells can be safely applied in an allogeneic setting, making them important effector cells for the development of off-the-shelf therapeutics for adoptive cancer immunotherapy. To further enhance their therapeutic potential, here, we engineered continuously expanding NK-92 cells as a clinically relevant model to express a humanized second-generation chimeric antigen receptor (CAR) with a composite CD28-CD3ζ signaling domain (hu14.18.28.z) that targets the disialoganglioside GD2, which is expressed at high levels by neuroblastoma cells and other tumors of neuroectodermal origin. In a separate approach, we fused an IL-15 superagonist (RD-IL15) to the GD2-CAR via a P2A processing site. Lentivirally transduced NK-92/hu14.18.28.z and NK-92/hu14.18.28.z_RD-IL15 cells both displayed high and stable CAR surface expression and specific cytotoxicity toward GD2-positive tumor cells. GD2-CAR NK cells carrying the RD-IL15 construct in addition expressed the IL-15 superagonist, resulting in self-enrichment and targeted cell killing in the absence of exogenous IL-2. Furthermore, co-culture with RD-IL15-secreting GD2-CAR NK cells markedly enhanced proliferation and cytotoxicity of bystander immune cells in a paracrine manner. Our results demonstrate that GD2-CAR NK cells co-expressing the IL-15 superagonist mediate potent direct and indirect antitumor effects, suggesting this strategy as a promising approach for the further development of functionally enhanced cellular therapeutics.

摘要翻译： 

与T淋巴细胞不同，自然杀伤（NK）细胞无需预先致敏即可在遭遇病毒感染或肿瘤细胞时迅速激活。此外，NK细胞可在同种异体环境中安全应用，这使其成为开发即用型过继性癌症免疫疗法的重要效应细胞。为进一步增强其治疗潜力，本研究以临床相关模型——可连续扩增的NK-92细胞为基础，通过基因工程改造使其表达人源化第二代嵌合抗原受体（CAR）。该受体采用复合型CD28-CD3ζ信号结构域（hu14.18.28.z），靶向神经母细胞瘤及其他神经外胚层起源肿瘤高表达的二唾液酸神经节苷脂GD2。通过另一技术路径，我们将IL-15超激动剂（RD-IL15）通过P2A切割位点与GD2-CAR融合表达。慢病毒转导的NK-92/hu14.18.28.z与NK-92/hu14.18.28.z_RD-IL15细胞均呈现高水平且稳定的CAR表面表达，并对GD2阳性肿瘤细胞具有特异性杀伤作用。携带RD-IL15结构的GD2-CAR NK细胞还能表达IL-15超激动剂，实现在无外源性IL-2条件下的自我增殖及靶向杀伤。此外，与分泌RD-IL15的GD2-CAR NK细胞共培养可通过旁分泌方式显著增强旁观免疫细胞的增殖与细胞毒性。研究结果表明，共表达IL-15超激动剂的GD2-CAR NK细胞能介导强效的直接与间接抗肿瘤效应，这为开发功能增强型细胞疗法提供了具有前景的新策略。

原文链接：

Co-Expression of an IL-15 Superagonist Facilitates Self-Enrichment of GD2-Targeted CAR-NK Cells and Mediates Potent Cell Killing in the Absence of IL-2