PO.ET02.08 · 实验与分子治疗
推动mRNA疗法迈向免疫肿瘤学应用：脂质纳米颗粒在正常小鼠和荷瘤小鼠中重复给药mRNA时展现出良好的安全性和有效性特征
Advancing mRNA therapeutics towards immuno-oncology applications: Lipid nanoparticles demonstrate favorable safety and efficacy profile with repeated mRNA administration in normal and tumor-bearing mice
作者与单位 Authors & Affiliations
摘要 Abstract
中文摘要
脂质纳米颗粒（LNPs）彻底革新了mRNA递送，为推进癌症免疫治疗提供了一种极具前景的方法。高效且安全地递送编码肿瘤抗原的mRNA能够激发强大的抗肿瘤免疫应答。在此，我们开发了用于免疫肿瘤学应用的LNP制剂，并通过递送编码卵清蛋白（OVA）的mRNA作为替代载荷来检验其适用性，同时在B16-F10-OVA异种移植黑色素瘤小鼠模型中评估其安全性、有效性和治疗潜力。通过对pKa、包封效率、流体动力学尺寸和生物分布特性进行合理筛选，选定了新型可电离脂质用于配制LNPs。编码OVA的mRNA采用NanoAssemblr™平台上的NxGen™微流控混合技术包封于LNPs中。对LNPs的流体动力学尺寸、多分散指数（PDI）、载药量和包封效率进行了表征。为评估长期安全性和稳定性，健康小鼠以21天为间隔多次肌肉注射（IM）LNP制剂。通过标准检测和临床观察评估理化稳定性和毒性。在此系列给药后，对B16-F10-OVA荷瘤小鼠肌肉注射接种载有OVA mRNA的LNPs。监测临床症状和生存结局，并与未治疗对照组进行比较。载有OVA mRNA的LNPs处于60至80 nm的理想尺寸范围内，PDI值低于0.2，包封效率大于95%。长期稳定性研究表明其保留了关键质量属性（CQAs）。在较长时间内重复肌肉注射该专有LNPs展现出稳定性和耐受性，在受试动物中未观察到可检测的毒性或不良反应。使用OVA mRNA-LNP疫苗治疗提高了荷瘤小鼠的生存率，与对照组相比显著延长了中位生存期。通过保留药物产品的CQAs，我们的新型LNP平台高效递送OVA mRNA，作为递送治疗性或免疫调节性核酸载荷的模型。此外，在较长时间内重复肌肉注射这些LNPs未导致可检测的不良事件，且OVA mRNA LNPs在B16-F10-OVA模型中诱导了强效的抗肿瘤效应。这种良好的安全性特征以及重复给药下的稳定性凸显了LNP适用于长期免疫调节或免疫肿瘤学应用的治疗策略。
查看英文原文 English abstract
Lipid nanoparticles (LNPs) have revolutionized mRNA delivery, offering a promising approach for advancing cancer immunotherapy. Effective and safe delivery of tumor antigen-encoding mRNA can stimulate robust anti-tumor immune responses. Here, we developed LNP formulations for immuno-oncology applications and tested its fit-for-use by delivering ovalbumin (OVA)-encoded mRNA as a surrogate payload and evaluated the safety, efficacy, and therapeutic potential in the B16-F10-OVA xenograft melanoma mouse model.Novel ionizable lipids were selected for formulating LNPs by rational screening of pKa, encapsulation efficiency, hydrodynamic size, and biodistribution properties. OVA-encoded mRNA was encapsulated in LNPs using NxGen™ microfluidic mixing technology on the NanoAssemblr™ platform. The LNPs were characterized for hydrodynamic size, polydispersity index (PDI), loading, and encapsulation efficiency. To assess long-term safety and stability, healthy mice received multiple intramuscular (IM) injections of the LNP formulations at 21-day intervals. Physicochemical stability and toxicity were evaluated through the standard assays and clinical observations. Following this series, B16-F10-OVA tumor-bearing mice were vaccinated intramuscularly with OVA mRNA-loaded LNPs. Clinical symptoms and survival outcomes were monitored and compared to untreated controls.OVA mRNA-loaded LNPs were in the desired size range of 60 to 80 nm with a PDI value below 0.2 and an encapsulation efficiency greater than 95%. Long-term stability studies indicated retention of critical quality attributes (CQAs). Repeated IM administration of the proprietary LNPs over an extended period demonstrated stability and tolerability with no detectable toxicity or adverse effects observed in treated animals. Treatment with the OVA mRNA-LNP vaccine improved survival in tumor-bearing mice, significantly extending median survival compared to controls.By retaining the CQAs of the drug products, our novel LNP platform efficiently delivers OVA mRNA as a model for delivering therapeutic or immune modulating nucleic acid payloads. Additionally, repeated IM administrations of these LNPs over an extended period resulted in no detectable adverse events and OVA mRNA LNPs induced potent anti-tumor effects in the B16-F10-OVA model. This favorable safety profile and stability upon repeated dosing underscore LNP suitability for long-term immune modulation or therapeutic strategies for immuno-oncology applications.
利益披露 Disclosure
S. Sadat, None.
A. Deyati,
Cytiva Employment.
K. Golla, None..
J. Paquette, None..
N. Aibani, None..
A. Thomas, None.