Vallera Daniel A, Ferrone Soldano, Kodal Behiye, Hinderlie Peter, Bendzick Laura, Ettestad Brianna, Hallstrom Caroline, Zorko Nicholas A, Rao Arpit, Fujioka Naomi, Ryan Charles J, Geller Melissa A, Miller Jeffrey S, Felices Martin
Masonic Cancer Center, Laboratory of Molecular Cancer Therapeutics, Department of Radiation Oncology, University of Minnesota, Minneapolis, MN 55455, USA.
Massachusetts General Hospital, Harvard Medical School, Boston, MA 02115, USA.
Cancers (Basel). 2020 Sep 18;12(9):2659. doi: 10.3390/cancers12092659.
We improved the bispecific antibody platform that primarily engages natural killer (NK) cells to kill cancer cells through antibody-dependent cellular cytotoxicity (ADCC) by adding IL-15 as a crosslinker that expands and self-sustains the effector NK cell population. The overall goal was to target B7-H3, an established marker predominantly expressed on cancer cells and minimally expressed on normal cells, and prove that it could target cancer cells in vitro and inhibit tumor growth in vivo. The tri-specific killer engager (TriKE) was assembled by DNA shuffling and ligation using DNA encoding a camelid anti-CD16 antibody fragment, a wild-type IL-15 moiety, and an anti-B7-H3 scFv (clone 376.96). The expressed and purified cam1615B7H3 protein was tested for in vitro NK cell activity against a variety of tumors and in vivo against a tagged human MA-148 ovarian cancer cell line grafted in NSG mice. cam1615B7H3 showed specific NK cell expansion, high killing activity across a range of B7-H3+ carcinomas, and the ability to mediate growth inhibition of aggressive ovarian cancer in vivo. cam1615B7H3 TriKE improves NK cell function, expansion, targeted cytotoxicity against various types of B7-H3-positive human cancer cell lines, and delivers an anti-cancer effect in vivo in a solid tumor setting.
我们改进了双特异性抗体平台，该平台主要通过抗体依赖性细胞毒性（ADCC）作用使自然杀伤（NK）细胞杀伤癌细胞，方法是添加白细胞介素-15（IL-15）作为交联剂，以扩增效应性NK细胞群体并使其自我维持。总体目标是靶向B7-H3，这是一种在癌细胞上大量表达而在正常细胞上极少表达的既定标志物，并证明其在体外可靶向癌细胞，在体内可抑制肿瘤生长。三特异性杀伤衔接子（TriKE）通过DNA改组和连接组装而成，使用编码骆驼科抗CD16抗体片段、野生型IL-15部分和抗B7-H3单链抗体片段（克隆376.96）的DNA。对表达并纯化的cam1615B7H3蛋白进行了测试，检测其针对多种肿瘤的体外NK细胞活性以及针对接种于NSG小鼠体内的标记人MA-148卵巢癌细胞系的体内活性。cam1615B7H3表现出特异性NK细胞扩增、对一系列B7-H3阳性癌的高杀伤活性以及在体内介导侵袭性卵巢癌生长抑制的能力。cam1615B7H3 TriKE改善了NK细胞功能、扩增能力、对各种类型B7-H3阳性人癌细胞系的靶向细胞毒性，并在实体瘤环境中在体内发挥抗癌作用。
