Dataset related to article "Arginine-Based Poly(I:C)-Loaded Nanocomplexes for the Polarization of Macrophages Toward M1-Antitumoral Effectors"

This record contains data related to article “Arginine-Based Poly(I:C)-Loaded Nanocomplexes for the Polarization of Macrophages Toward M1-Antitumoral Effectors" Abstract: <strong>Background:</strong> Tumor-associated macrophages (TAMs), with M2-like immunosuppressive profiles, are key players in the development and dissemination of tumors. Hence, the induction of M1 pro-inflammatory and anti-tumoral states is critical to fight against cancer cells. The activation of the endosomal toll-like receptor 3 by its agonist poly(I:C) has shown to efficiently drive this polarization process. Unfortunately, poly(I:C) presents significant systemic toxicity, and its clinical use is restricted to a local administration. Therefore, the objective of this work has been to facilitate the delivery of poly(I:C) to macrophages through the use of nanotechnology, that will ultimately drive their phenotype toward pro-inflammatory states. <strong>Methods:</strong> Poly(I:C) was complexed to arginine-rich polypeptides, and then further enveloped with an anionic polymeric layer either by film hydration or incubation. Physicochemical characterization of the nanocomplexes was conducted by dynamic light scattering and transmission electron microscopy, and poly(I:C) association efficiency by gel electrophoresis. Primary human-derived macrophages were used as relevant <em>in vitro</em> cell model. Alamar Blue assay, ELISA, PCR and flow cytometry were used to determine macrophage viability, polarization, chemokine secretion and uptake of nanocomplexes. The cytotoxic activity of pre-treated macrophages against PANC-1 cancer cells was assessed by flow cytometry. <strong>Results:</strong> The final poly(I:C) nanocomplexes presented sizes lower than 200 nm, with surface charges ranging from +40 to -20 mV, depending on the envelopment. They all presented high poly(I:C) loading values, from 12 to 50%, and great stability in cell culture media. <em>In vitro</em>, poly(I:C) nanocomplexes were highly taken up by macrophages, in comparison to the free molecule. Macrophage treatment with these nanocomplexes did not reduce their viability and efficiently stimulated the secretion of the T-cell recruiter chemokines CXCL10 and CCL5, of great importance for an effective anti-tumor immune response. Finally, poly(I:C) nanocomplexes significantly increased the ability of treated macrophages to directly kill cancer cells. <strong>Conclusion:</strong> Overall, these enveloped poly(I:C) nanocomplexes might represent a therapeutic option to fight cancer through the induction of cytotoxic M1-polarized macrophages.本数据集收录与论文《基于精氨酸的聚肌胞苷酸（poly(I:C)）负载纳米复合物用于将巨噬细胞极化为M1型抗肿瘤效应细胞》相关的数据。摘要：<strong>背景：</strong>肿瘤相关巨噬细胞（tumor-associated macrophages, TAMs）具有M2样免疫抑制表型，是肿瘤发生与扩散的关键介导因素。因此，诱导巨噬细胞向M1型促炎、抗肿瘤表型极化，是对抗肿瘤细胞的核心策略。胞内体Toll样受体3（toll-like receptor 3, TLR3）可通过其激动剂聚肌胞苷酸（poly(I:C)）激活，已被证实可有效驱动该极化过程。然而，聚肌胞苷酸存在显著的全身毒性，临床应用仅局限于局部给药。因此，本研究旨在借助纳米技术实现聚肌胞苷酸向巨噬细胞的靶向递送，最终促使巨噬细胞向促炎表型转化。<strong>方法：</strong>将聚肌胞苷酸与富含精氨酸的多肽复合，随后通过薄膜水化法或孵育法进一步用阴离子聚合物层包覆。采用动态光散射与透射电子显微镜对纳米复合物进行理化表征，通过凝胶电泳测定聚肌胞苷酸的包封效率。以原代人源巨噬细胞作为合适的体外（in vitro）细胞模型。采用阿尔玛蓝（Alamar Blue）检测法、酶联免疫吸附试验（ELISA）、聚合酶链式反应（PCR）与流式细胞术分别检测巨噬细胞的活力、极化状态、趋化因子分泌水平以及纳米复合物的摄取情况。通过流式细胞术评估经预处理的巨噬细胞对PANC-1癌细胞的细胞毒活性。<strong>结果：</strong>最终制备的聚肌胞苷酸纳米复合物粒径均小于200 nm，表面电荷范围为+40 mV至-20 mV，具体数值取决于包覆工艺。所有纳米复合物均具有较高的聚肌胞苷酸负载率（12%~50%），且在细胞培养基中稳定性良好。体外实验表明，与游离聚肌胞苷酸相比，纳米复合物可被巨噬细胞高效摄取。经纳米复合物处理的巨噬细胞未出现活力下降，且可有效刺激T细胞招募趋化因子CXCL10与CCL5的分泌，这两类趋化因子对高效抗肿瘤免疫应答至关重要。最后，聚肌胞苷酸纳米复合物可显著提升巨噬细胞直接杀伤癌细胞的能力。<strong>结论：</strong>综上，这类经包覆的聚肌胞苷酸纳米复合物有望成为通过诱导细胞毒性M1型极化巨噬细胞对抗癌症的新型治疗方案。