Chikileva Irina O, Bruter Alexandra V, Persiyantseva Nadezhda A, Zamkova Maria A, Vlasenko Raimonda Ya, Dolzhikova Yuliya I, Shubina Irina Zh, Donenko Fedor V, Lebedinskaya Olga V, Sokolova Darina V, Pokrovsky Vadim S, Fedorova Polina O, Ustyuzhanina Nadezhda E, Anisimova Natalia Yu, Nifantiev Nikolay E, Kiselevskiy Mikhail V
Research Institute of Experimental Therapy and Diagnostics of Tumor, NN Blokhin National Medical Center of Oncology, 115478 Moscow, Russia.
Center for Precision Genome Editing and Genetic Technologies for Biomedicine, Institute of Gene Biology, Russian Academy of Sciences, 119334 Moscow, Russia.
Biomedicines. 2023 Sep 18;11(9):2563. doi: 10.3390/biomedicines11092563.
Human epidermal growth factor receptor 2 (HER2) is overexpressed in numerous cancer cell types. Therapeutic antibodies and chimeric antigen receptors (CARs) against HER2 were developed to treat human tumors. The major limitation of anti-HER2 CAR-T lymphocyte therapy is attributable to the low HER2 expression in a wide range of normal tissues. Thus, side effects are caused by CAR lymphocyte "on-target off-tumor" reactions. We aimed to develop safer HER2-targeting CAR-based therapy. CAR constructs against HER2 tumor-associated antigen (TAA) for transient expression were delivered into target T and natural killer (NK) cells by an effective and safe non-viral transfection method via nucleofection, excluding the risk of mutations associated with viral transduction. Different in vitro end-point and real-time assays of the CAR lymphocyte antitumor cytotoxicity and in vivo human HER2-positive tumor xenograft mice model proved potent cytotoxic activity of the generated CAR-T-NK cells. Our data suggest transient expression of anti-HER2 CARs in plasmid vectors by human lymphocytes as a safer treatment for HER2-positive human cancers. We also conducted preliminary investigations to elucidate if fucosylated chondroitin sulfate may be used as a possible agent to decrease excessive cytokine production without negative impact on the CAR lymphocyte antitumor effect.
人表皮生长因子受体2（HER2）在多种癌细胞类型中过表达。针对HER2开发了治疗性抗体和嵌合抗原受体（CAR）来治疗人类肿瘤。抗HER2嵌合抗原受体修饰的T淋巴细胞（CAR-T）疗法的主要局限性在于广泛的正常组织中HER2表达较低。因此，副作用是由CAR淋巴细胞的“靶向非肿瘤”反应引起的。我们旨在开发更安全的基于HER2靶向CAR的疗法。通过核转染这种有效且安全的非病毒转染方法，将针对HER2肿瘤相关抗原（TAA）的CAR构建体递送至靶T细胞和自然杀伤（NK）细胞中，排除了与病毒转导相关的突变风险。对CAR淋巴细胞抗肿瘤细胞毒性进行的不同体外终点和实时检测以及体内人HER2阳性肿瘤异种移植小鼠模型证明了所产生的CAR-T-NK细胞具有强大的细胞毒性活性。我们的数据表明，人淋巴细胞在质粒载体中瞬时表达抗HER2 CAR可作为HER阳性人类癌症的更安全治疗方法。我们还进行了初步研究，以阐明岩藻糖基化硫酸软骨素是否可作为一种可能的药物来减少过量的细胞因子产生，而不会对CAR淋巴细胞的抗肿瘤作用产生负面影响。
