Safe and effective in vivo delivery of DNA and RNA using proteolipid vehicles

Affiliations

• Department of Oncology, University of Alberta, Edmonton, AB T6G 2E1, Canada
• Entos Pharmaceuticals, 10230 Jasper Avenue, Suite 4550, Edmonton, AB T5J 4P6, Canada
• OncoSenX, 701 Fifth Avenue, Suite 4200, Seattle, WA 98104, USA
• Department of Microbiology & Immunology, Dalhousie University, Halifax, NS B3H 4R2, Canada
• Oisin Biotechnologies, 701 Fifth Avenue, Suite 4200, Seattle, WA 98104, USA

PMID:  #  DOI: 10.1016/j.cell.2024.07.023

Abstract

Genetic medicines show promise for treating various diseases, yet clinical success has been limited bytolerability, scalability, and immunogenicity issues of current delivery platforms. To overcome these, we developed a proteolipid vehicle (PLV) by combining features from viral and non-viral approaches. PLVs incorporate fusion-associated small transmembrane (FAST) proteins isolated from fusogenic orthoreoviruses into a well-tolerated lipid formulation, using scalable microfluidic mixing. Screening a FAST protein library, we identified a chimeric FAST protein with enhanced membrane fusion activity that improved gene expression from an optimized lipid formulation. Systemically administered FAST-PLVs showed broad biodistribution and effective mRNA and DNA delivery in mouse and non-human primate models. FAST-PLVs show low immunogenicity and maintain activity upon repeat dosing. Systemic administration of follistatin DNA gene therapy with FAST-PLVs raised circulating follistatin levels and significantly increased muscle mass and grip strength. These results demonstrate the promising potential of FAST-PLVs for redosable gene therapies and genetic medicines.

核酸药物在治疗多种疾病方面显示出巨大潜力，但由于现有递送平台在耐受性、可扩展性和免疫原性方面存在问题，临床成功率有限。为了解决这些问题，我们结合了病毒和非病毒方法的特点,开发了一种蛋白脂质载体（proteolipid vehicle, PLV）。PLV将正呼肠孤病毒中分离出的融合相关小跨膜蛋白整合到一种耐受性良好的脂质体中，并使用可扩展的微流体混合技术进行制备。通过筛选FAST蛋白库，我们鉴定出一种具有增强膜融合活性的嵌合FAST蛋白，能够提高优化后的脂质配方的基因表达效果。系统性给药的FAST-PLV在小鼠和非人灵长类动物模型中显示出广泛的体内分布，并有效递送mRNA和DNA。FAST-PLV的免疫原性较低，并且在重复给药时保持活性。使用FAST-PLV进行肌肉抑素DNA基因治疗，通过系统给药提高了循环中的卵泡抑素水平，显著增加了肌肉质量和握力。以上结果表明，FAST-PLV在可重复给药的基因疗法和核酸药物中展现了广阔的应用前景。