Moiseenko Alena, Sinadinos Anthony, Sergijenko Ana, Pineault Kyriel, Saleh Aarash, Nekola Konradin, Strang Nathalie, Eleftheraki Anastasia, Boyd A Christopher, Davies Jane C, Gill Deborah R, Hyde Stephen C, McLachlan Gerry, Rath Tim, Rothe Michael, Schambach Axel, Hobbie Silke, Schuler Michael, Maier Udo, Thomas Matthew J, Mennerich Detlev, Schmidt Manfred, Griesenbach Uta, Alton Eric W F W, Kreuz Sebastian
Boehringer Ingelheim Pharma GmbH, Biberach an der Riss, Germany.
UK Respiratory Gene Therapy Consortium, London, UK.
Eur Respir J. 2025 Jan 30;65(1). doi: 10.1183/13993003.01683-2023. Print 2025 Jan.
Cystic fibrosis (CF) is caused by mutations in the CF transmembrane conductance regulator (CFTR) gene. CFTR modulators offer significant improvements, but ∼10% of patients remain nonresponsive or are intolerant. This study provides an analysis of rSIV.F/HN, a lentiviral vector optimised for lung delivery, including CFTR protein expression, functional correction of CFTR defects and genomic integration site analysis in preparation for a first-in-human clinical trial.
Air-liquid interface cultures of primary human bronchial epithelial cells (HBECs) from CF patients (F508del/F508del), as well as a CFTR-deficient immortalised human lung epithelial cell line mimicking class I (CFTR-null) homozygous mutations, were used to assess transduction efficiency. Quantification methods included a novel proximity ligation assay for CFTR protein expression. For assessment of CFTR channel activity, Ussing chamber studies were conducted. The safety profile was assessed using integration site analysis and insertional mutagenesis studies.
rSIV.F/HN expressed CFTR and restored CFTR-mediated chloride currents to physiological levels in primary F508del/F508del HBECs as well as in a class I cells. In contrast, the latter could not be achieved by small-molecule CFTR modulators, underscoring the potential of gene therapy for this mutation class. Combination of rSIV.F/HN-CFTR with the potentiator ivacaftor showed a greater than additive effect. The genomic integration pattern showed no site predominance (frequency of occurrence ≤10%), and a low risk of insertional mutagenesis was observed in an immortalisation assay.
The results underscore rSIV.F/HN as a promising gene therapy vector for CF, providing a mutation-agnostic treatment option.
囊性纤维化（CF）由囊性纤维化跨膜传导调节因子（CFTR）基因突变引起。CFTR调节剂带来了显著改善，但约10%的患者仍无反应或不耐受。本研究对rSIV.F/HN进行了分析，rSIV.F/HN是一种为肺部递送而优化的慢病毒载体，包括CFTR蛋白表达、CFTR缺陷的功能校正以及基因组整合位点分析，为首次人体临床试验做准备。
使用来自CF患者（F508del/F508del）的原代人支气管上皮细胞（HBECs）的气液界面培养物，以及模拟I类（CFTR缺失）纯合突变的CFTR缺陷永生化人肺上皮细胞系，评估转导效率。定量方法包括一种用于CFTR蛋白表达的新型邻近连接测定法。为评估CFTR通道活性，进行了尤斯灌流小室研究。使用整合位点分析和插入诱变研究评估安全性。
rSIV.F/HN在原代F508del/F508del HBECs以及I类细胞中表达CFTR，并将CFTR介导的氯离子电流恢复到生理水平。相比之下，小分子CFTR调节剂无法实现后者，突出了基因疗法针对此类突变的潜力。rSIV.F/HN-CFTR与增强剂依伐卡托联合使用显示出大于相加的效果。基因组整合模式未显示位点优势（出现频率≤10%），并且在永生化测定中观察到插入诱变的低风险。
结果强调rSIV.F/HN作为一种有前景的CF基因治疗载体，提供了一种不依赖突变的治疗选择。
