Ruzanova Vera, Proskurina Anastasia, Ritter Genrikh, Dolgova Evgeniya, Oshikhmina Sofya, Kirikovich Svetlana, Levites Evgeniy, Efremov Yaroslav, Taranov Oleg, Ostanin Alexandr, Chernykh Elena, Kolchanov Nikolay, Bogachev Sergey
Laboratory of Induced Cellular Processes, Institute of Cytology and Genetics SB RAS, Novosibirsk, 630090, Russia.
Novosibirsk National Research State University, Novosibirsk, 630090, Russia.
Oncol Res. 2025 Apr 18;33(5):1229-1248. doi: 10.32604/or.2025.059411. eCollection 2025.
Currently, there exist two approaches to the treatment of malignant neoplasms: the Karanahan technology and vaccination, which are based on chronometric delivery of therapeutic agents to the tumor depending on the characteristics of tumor cells, as well as the immune status. The main purpose of this study was to experimentally prove the feasibility of combining the Karanahan technology and vaccination with αOX40 antibodies into a single therapeutic platform to achieve a potent additive antitumor therapeutic effect.
BALB/c mice grafted with B-cellular lymphoma A20 were treated using the Karanahan technology consisting of intraperitoneal cyclophosphamide administrations and intratumoral DNA injections according to an individually determined therapeutic regimen, together with vaccination with αOX40. A pathomorphological analysis of the organs of experimental animals that died during the initial attempt to combine the two technologies was carried out. An analysis of blood cell populations was performed to determine the safe time for antibody administration: the number of immune cells capable of activating systemic inflammation (CD11b+Ly-6C+, CD11b+Ly-6G+, CD3-NKp46+CD11b+), the presence of Fc receptor and OX40 on the surface of these cells, and the number of neutrophils activated to NETosis were analyzed. Based on the analysis results, the antitumor efficacy of various modes of combining the Karanahan technology and vaccination was studied.
When αOX40 was administered 5 h after each treatment using the Karanahan technology, mass death of mice caused by systemic inflammation and multiple organ failure was observed. The state of blood cells after the treatment using the Karanahan technology at the time points corresponding to antibody injections was analyzed to elucidate the reasons for this effect. It was found that at some time points, there occurs activation of the immune system and a powerful release (up to 16%) of monocytes and granulocytes carrying Fc receptor and OX40 on their surface into blood; when interacting with αOX40, they can activate the lytic potential of these cells. Activation of neutrophils to NETosis was also observed. Based on these findings, a study was carried out in different time regimes to combine the Karanahan technology and αOX40 injections. When αOX40 was injected into the points of minimal release of myeloid cells into the blood, increased survival rate and the greatest antitumor efficacy were observed: 37% of animals survived without relapses on day 100 after experiment initiation. The results obtained indicate that it is possible to combine the Karanahan technology and vaccination with αOX40, with obligatory constant monitoring of the number of myeloid cells in peripheral blood to determine the safe time for antibody injection.
目前，恶性肿瘤的治疗存在两种方法：卡拉纳汉技术和疫苗接种，这两种方法基于根据肿瘤细胞特征以及免疫状态向肿瘤定时递送治疗药物。本研究的主要目的是通过实验证明将卡拉纳汉技术、疫苗接种与αOX40抗体结合到一个单一治疗平台以实现强效的附加抗肿瘤治疗效果的可行性。
对移植了B细胞淋巴瘤A20的BALB/c小鼠，按照个体确定的治疗方案，采用由腹腔注射环磷酰胺和瘤内注射DNA组成的卡拉纳汉技术进行治疗，并同时接种αOX40疫苗。对在最初尝试将两种技术结合时死亡的实验动物的器官进行病理形态学分析。进行血细胞群体分析以确定抗体给药的安全时间：分析能够激活全身炎症的免疫细胞数量（CD11b+Ly-6C+、CD11b+Ly-6G+、CD3-NKp46+CD11b+）、这些细胞表面Fc受体和OX40的存在情况以及被激活发生中性粒细胞胞外陷阱形成的中性粒细胞数量。基于分析结果，研究了卡拉纳汉技术与疫苗接种不同组合方式的抗肿瘤疗效。
在每次使用卡拉纳汉技术治疗后5小时给予αOX40时，观察到小鼠因全身炎症和多器官衰竭而大量死亡。分析在与抗体注射对应的时间点使用卡拉纳汉技术治疗后血细胞的状态，以阐明这种效应的原因。发现在某些时间点，免疫系统会被激活，并且表面携带Fc受体和OX40的单核细胞和粒细胞会大量释放（高达16%）到血液中；当它们与αOX40相互作用时，可激活这些细胞的溶解潜能。还观察到中性粒细胞被激活发生中性粒细胞胞外陷阱形成。基于这些发现，在不同时间方案下进行了将卡拉纳汉技术与αOX40注射相结合的研究。当在骨髓细胞向血液中释放最少的时间点注射αOX40时，观察到存活率提高且抗肿瘤疗效最佳：在实验开始后第100天，37%的动物存活且无复发。所得结果表明可以将卡拉纳汉技术、疫苗接种与αOX40相结合，但必须持续监测外周血中骨髓细胞数量以确定抗体注射的安全时间。
