肿瘤术中成像的患者衍生细胞外囊泡的临床前药理学
Preclinical pharmacology of patient-derived extracellular vesicles for the intraoperative imaging of tumors
影响因子:13.30000
分区:医学1区 Top / 医学：研究与实验1区
发表日期:2024
作者:
Alessandro Villa, Daniela Crescenti, Zemira De Mitri, Elisabetta Crippa, Silvia Rosa, Nicoletta Rizzi, Fereshteh Shojaei-Ghahrizjani, Monica Rebecchi, Simona Vincenti, Francesca Selmin, Electra Brunialti, Nicolò Simonotti, Marianna Maspero, Michele Dei Cas, Camilla Recordati, Saverio Paltrinieri, Alessia Giordano, Rita Paroni, Margherita Galassi, Vito Ladisa, Flavio Arienti, Francesco Cilurzo, Vincenzo Mazzaferro, Paolo Ciana
摘要
与健康个体不同的肿瘤患者血浆衍生的细胞外囊泡（EV）具有明显的肿瘤靶向特性。我们先前已经表明，与患者衍生的细胞外囊泡（PDEV）配制了近红外（NIR）荧光染料氨基氨基绿（ICG），以选择性递送到肿瘤组织。该染色方案有望在术中肿瘤边缘成像中临床应用，从而实现精确的肿瘤组织切除。为此，我们提出了Oncogreen方案，涉及PDEV隔离，ICG加载和重新灌注相同患者。方法：通过对小鼠的体内研究，我们概述了术中肿瘤成像，PDEV生物分布动力学和潜在治疗相关的毒性效应的PDEVS-ICG的关键药理参数。此外，我们建立了一种基于血浆置换的方案，用于分离自体PDEV，以确保对人类治疗的必要大规模剂量。还探索了一种潜在的基于冻干的保存方法，以促进PDEV的存储和运输。结果：该研究确定了清晰的术中肿瘤边缘成像所需的PDEVS-ICG的有效剂量。 PDEV的生物分布动力学表现出对肿瘤组织的有利靶向，而没有靶向分布。毒理学评估显示与治疗无关。基于血浆的隔离方案成功得出了足够数量的自体PDEV，而冻干保存方法维持了PDEV的功能完整性，以便随后的临床应用。结论：我们的研究为自体PDEV的直接临床应用奠定了基础，最初侧重于术中成像。利用自体PDEV有可能加速电动汽车作为抗肿瘤剂到癌组织的靶向输送工具的整合。这种方法有望提高肿瘤组织切除的精度，并改善肿瘤患者的整体手术结果。
Abstract
Extracellular vesicles (EVs) derived from the plasma of oncological patients exhibit significant tumor-targeting properties, unlike those from healthy individuals. We have previously shown the feasibility of formulating the near-infrared (NIR) fluorescent dye indocyanine green (ICG) with patient-derived extracellular vesicles (PDEVs) for selective delivery to neoplastic tissue. This staining protocol holds promise for clinical application in intraoperative tumor margin imaging, enabling precise neoplastic tissue resection. To this end, we propose the ONCOGREEN protocol, involving PDEV isolation, ICG loading, and reinfusion into the same patients. Methods: By in vivo studies on mice, we outlined key pharmacological parameters of PDEVs-ICG for intraoperative tumor imaging, PDEV biodistribution kinetics, and potential treatment-related toxicological effects. Additionally, we established a plasmapheresis-based protocol for isolating autologous PDEVs, ensuring the necessary large-scale dosage for human treatment. A potential lyophilization-based preservation method was also explored to facilitate the storage and transport of PDEVs. Results: The study identified the effective dose of PDEVs-ICG necessary for clear intraoperative tumor margin imaging. The biodistribution kinetics of PDEVs showed favorable targeting to neoplastic tissues, without off-target distribution. Toxicological assessments revealed no significant adverse effects associated with the treatment. The plasmapheresis-based isolation protocol successfully yielded a sufficient quantity of autologous PDEVs, and the lyophilization preservation method maintained the functional integrity of PDEVs for subsequent clinical application. Conclusions: Our research lays the groundwork for the direct clinical application of autologous PDEVs, initially focusing on intraoperative imaging. Utilizing autologous PDEVs has the potential to accelerate the integration of EVs as a targeted delivery tool for anti-neoplastic agents to cancerous tissue. This approach promises to enhance the precision of neoplastic tissue resection and improve overall surgical outcomes for oncological patients.