OX40配体的非病毒介导的基因转移用于肿瘤免疫疗法

免疫检查点封锁（ICB）正在迅速成为许多癌症治疗的护理标准。但是，对这种类型的治疗反应的患者子集有限。促进抗肿瘤免疫力的另一种方法是使用免疫刺激分子，例如细胞因子或T细胞共刺激剂。免疫疗法的系统给药会导致与免疫相关的显着不良事件（IRAE），因此需要局部抗肿瘤作用。实现这一目标的一种方法是肿瘤内非病毒基因免疫治疗，它允许长时间和局部基因表达和多次药物给药。在这项研究中，我们将先前描述的非病毒基因递送系统PEG-PEI-TAT共聚物PPT与鼠OX40L编码质粒DNA结合在一起。所得的OX40L/PPT纳米颗粒的表征是通过凝胶动弹性测定，动态光散射分析和VITRO TREFRO EFFROFICE评估来表征的。使用皮下（S.C.）植入的CT26（结肠癌），B16F0（黑色素瘤）和4T1（乳腺癌）肿瘤模型估算肿瘤内（I.T.）施用的纳米颗粒的抗肿瘤功效。使用流式细胞术分析基质免疫细胞群体的动力学。减肥和恶病质被用作IRAE指标。 I.T.组合的影响S.C.估计了带有腹膜内PD-1 ICB的OX40L/PPT。 CT26肿瘤模型。获得的OX40L/PPT纳米颗粒具有适用于细胞转染的特性，并在所有三个研究的癌症模型中都提供了OX40L蛋白表达。我们观察到OX40L/PPT治疗成功抑制了B16F0和CT26肿瘤模型的肿瘤生长，并显示出抑制4T1肿瘤生长的趋势。在B16F0肿瘤模型中，OX40L/PPT治疗导致抗肿瘤效应的NK和T杀伤细胞的增加，并减少肿瘤基质中的肿瘤髓样细胞种群的减少。在所有3个肿瘤模型中均未观察到IRAE迹象，这表明小鼠的治疗耐受性良好。 Combining OX40L/PPT with PD-1 ICB significantly improved treatment efficacy in the CT26 subcutaneous colon cancer model, providing protective immunity against CT26 colon cancer cells.Overall, the anti-tumor efficacy observed with OX40L non-viral gene therapy, whether administered alone or in combination with ICB, highlights its potential to revolutionize cancer gene therapy, thus paving the way对于癌症治疗领域的前所未有的进步。

Immune checkpoint blockade (ICB) is rapidly becoming a standard of care in the treatment of many cancer types. However, the subset of patients who respond to this type of therapy is limited. Another way to promote antitumoral immunity is the use of immunostimulatory molecules, such as cytokines or T cell co-stimulators. The systemic administration of immunotherapeutics leads to significant immune-related adverse events (irAEs), therefore, the localized antitumoral action is needed. One way to achieve this is intratumoral non-viral gene-immune therapy, which allows for prolonged and localized gene expression, and multiple drug administration. In this study, we combined the previously described non-viral gene delivery system, PEG-PEI-TAT copolymer, PPT, with murine OX40L-encoding plasmid DNA.The resulting OX40L/PPT nanoparticles were characterized via gel mobility assay, dynamic light scattering analysis and in vitro transfection efficiency evaluation. The antitumoral efficacy of intratumorally (i.t.) administered nanoparticles was estimated using subcutaneously (s.c.) implanted CT26 (colon cancer), B16F0 (melanoma) and 4T1 (breast cancer) tumor models. The dynamics of stromal immune cell populations was analyzed using flow cytometry. Weight loss and cachexia were used as irAE indicators. The effect of combination of i.t. OX40L/PPT with intraperitoneal PD-1 ICB was estimated in s.c. CT26 tumor model.The obtained OX40L/PPT nanoparticles had properties applicable for cell transfection and provided OX40L protein expression in vitro in all three investigated cancer models. We observed that OX40L/PPT treatment successfully inhibited tumor growth in B16F0 and CT26 tumor models and showed a tendency to inhibit 4T1 tumor growth. In B16F0 tumor model, OX40L/PPT treatment led to the increase in antitumoral effector NK and T killer cells and to the decrease in pro-tumoral myeloid cells populations within tumor stroma. No irAE signs were observed in all 3 tumor models, which indicates good treatment tolerability in mice. Combining OX40L/PPT with PD-1 ICB significantly improved treatment efficacy in the CT26 subcutaneous colon cancer model, providing protective immunity against CT26 colon cancer cells.Overall, the anti-tumor efficacy observed with OX40L non-viral gene therapy, whether administered alone or in combination with ICB, highlights its potential to revolutionize cancer gene therapy, thus paving the way for unprecedented advancements in the cancer therapy field.