WS05.02CFTR mRNA delivery with a revolutionary non-LNP nanoemulsion formulation to differentiated primary human airway epithelium and airway organoid

IF 5.4 2区医学 Q1 RESPIRATORY SYSTEM

Journal of Cystic Fibrosis Pub Date : 2025-06-01 DOI:10.1016/j.jcf.2025.03.517

K. Ito , S. Hassibi , Y. Yamamoto , J. Shur , G. Rapeport , S. Sobolov

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引用次数: 0

Abstract

Objectives

Cystic fibrosis (CF) is a progressive genetic disorder with impaired normal clearance of mucus due to the defect of a chloride channel, cystic fibrosis transmembrane regulator (CFTR). Currently approved CFTR modulators are applicable to limited genotypes and the universally applicable therapy would be functional CFTR protein expression by CFTR gene delivery. To address this challenge, RIGImmune has developed a non-LNP formulation delivery system, Nano-Emulsion Enhanced Delivery (NEED^TM) platform to deliver optimised CFTR encoded mRNA therapeutics.

Methods

Optimised CFTR mRNAs (Northern RNA, Canada) formulated with NEED^TM nanoemulsion system (RIG-301) were apically applied for up to 4 hrs to air-liquid interface (ALI) cultured differentiated human alveolar or bronchial epithelium (hAE or hBE, Epithelix) obtained from healthy subjects. Cells were collected at different timepoints up to 72hrs post-delivery, and CFTR protein was detected by western blotting. RIG-301 was also applied to human induced pluripotent stem cell (iPSC) derived CF ΔF508- airway organoids, and effects of forskolin-induced swelling were evaluated (HiLung Inc. Japan).

Results

RIG-301 nanoemulsion system (z-average particle size 176.9 nm) showed higher and long-lasting CFTR protein expression than naked CFTR mRNA delivery or delivered with jetPrime transfection reagent in hAE and hBE. In addition, RIG-301 showed an increased forskolin-induced swelling of CF-airway organoid, and the effects were comparable to or better than that of a triple combinations of CFTR modulators (VX-661/VX-445/VX-770: 3, 3 and 1 µM, respectively).

Conclusion

Our results demonstrate the capability of the NEED^TM platform to deliver optimised CFTR mRNA in ALI hAE and hBE cultures, as well as CF organoid. These preclinical data warrant further investigation of CFTR delivery and functional rescue in cells derived from CF patients.

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通过革命性的非lnp纳米乳配方将cftr mRNA递送至分化的人气道上皮和气道类器官

囊性纤维化（CF）是一种进行性遗传性疾病，由于氯离子通道、囊性纤维化跨膜调节剂（CFTR）的缺陷而导致粘液正常清除受损。目前批准的CFTR调节剂适用于有限的基因型，普遍适用的治疗方法将是通过CFTR基因递送来表达功能性CFTR蛋白。为了应对这一挑战，RIGImmune开发了一种非lnp配方递送系统，纳米乳液增强递送（NEEDTM）平台，用于递送优化的CFTR编码mRNA疗法。方法采用NEEDTM纳米乳体系（RIG-301）配制的CFTR mrna (Northern RNA, Canada)，将优化后的CFTR mrna顶端应用于健康受试者的分化人肺泡或支气管上皮（hAE或hBE， epithelial）的气液界面（ALI）培养，时间长达4小时。在分娩后72h的不同时间点收集细胞，用western blotting检测CFTR蛋白。RIG-301也应用于人诱导多能干细胞（iPSC）衍生的CF ΔF508-气道类器官，并评估了福斯克林诱导肿胀的效果（HiLung Inc.）。日本)。结果rig -301纳米乳体系（z-平均粒径为176.9 nm）在hAE和hBE中的CFTR蛋白表达量高于裸递送或jetPrime转染试剂递送。此外，RIG-301显示福斯克林诱导的cf -气道类器官肿胀增加，其效果与CFTR调节剂三种组合（VX-661/VX-445/VX-770：分别为3、3和1µM）相当或更好。我们的研究结果表明，NEEDTM平台能够在ALI hAE和hBE培养以及CF类器官中传递优化的CFTR mRNA。这些临床前数据为进一步研究CFTR在CF患者细胞中的传递和功能挽救提供了依据。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Journal of Cystic Fibrosis 医学-呼吸系统

CiteScore

10.10

自引率

13.50%

发文量

1361

审稿时长

50 days

期刊介绍： The Journal of Cystic Fibrosis is the official journal of the European Cystic Fibrosis Society. The journal is devoted to promoting the research and treatment of cystic fibrosis. To this end the journal publishes original scientific articles, editorials, case reports, short communications and other information relevant to cystic fibrosis. The journal also publishes news and articles concerning the activities and policies of the ECFS as well as those of other societies related the ECFS.