Preparation for a first-in-man lentivirus trial in patients with cystic fibrosis.
Preparation for a first-in-man lentivirus trial in patients with cystic fibrosis.
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DOI：
10.1136/thoraxjnl-2016-208406
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发表时间：
2017-02
期刊：
影响因子：
10
通讯作者：
Zhu J
中科院分区：
文献类型：
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作者：
Alton EW;Beekman JM;Boyd AC;Brand J;Carlon MS;Connolly MM;Chan M;Conlon S;Davidson HE;Davies JC;Davies LA;Dekkers JF;Doherty A;Gea-Sorli S;Gill DR;Griesenbach U;Hasegawa M;Higgins TE;Hironaka T;Hyndman L;McLachlan G;Inoue M;Hyde SC;Innes JA;Maher TM;Moran C;Meng C;Paul-Smith MC;Pringle IA;Pytel KM;Rodriguez-Martinez A;Schmidt AC;Stevenson BJ;Sumner-Jones SG;Toshner R;Tsugumine S;Wasowicz MW;Zhu J
我们最近发现，非病毒基因治疗可以稳定囊性纤维化（CF）患者的肺功能下降。然而，效果是温和的，仍然需要更有效的基因转移剂。在临床前模型中，融合蛋白（F）/血凝素/神经氨酸酶蛋白（HN）假型慢病毒载体比非病毒载体更有效地用于肺基因转移。在使用慢病毒载体进行首次人体CF试验的准备过程中，我们进行了关键的转化临床前研究。在小鼠和人气液界面（ALI）培养物中评估携带一系列启动子/增强子元件的符合监管规定的载体，以选择主要候选者;使用该主要候选载体在CF模型中评估囊性纤维化跨膜传导受体（CFTR）表达和功能。毒性进行了评估，并对最近在IIb期临床试验中使用的领先的非病毒制剂进行了“基准”。绘制整合位点图谱，并确定转导效率以告知临床试验剂量范围。评估了预先存在的和获得性免疫对载体和几种临床相关输送器械中载体稳定性的影响。由延伸因子1α启动子和巨细胞病毒增强子组成的不含胞嘧啶鸟嘌呤二核苷酸（CpG）的杂合启动子/延伸因子1 α启动子（hCEF）在鼠肺和人ALI培养物中最有效（均高于背景至少2-log数量级）。功效（至少14%的气道细胞被转导）、毒性和整合位点特征支持进一步进展至临床试验，并且预先存在的和获得性免疫应答不干扰载体功效。前导rSIV.F/HN候选物表达功能性CFTR，并且载体在临床相关递送装置中保持90-100%的转导效率。这些数据支持F/HN假型慢病毒载体在2017年进入首次人体CF试验。
We have recently shown that non-viral gene therapy can stabilise the decline of lung function in patients with cystic fibrosis (CF). However, the effect was modest, and more potent gene transfer agents are still required. Fuson protein (F)/Hemagglutinin/Neuraminidase protein (HN)-pseudotyped lentiviral vectors are more efficient for lung gene transfer than non-viral vectors in preclinical models. In preparation for a first-in-man CF trial using the lentiviral vector, we have undertaken key translational preclinical studies. Regulatory-compliant vectors carrying a range of promoter/enhancer elements were assessed in mice and human air–liquid interface (ALI) cultures to select the lead candidate; cystic fibrosis transmembrane conductance receptor (CFTR) expression and function were assessed in CF models using this lead candidate vector. Toxicity was assessed and ‘benchmarked’ against the leading non-viral formulation recently used in a Phase IIb clinical trial. Integration site profiles were mapped and transduction efficiency determined to inform clinical trial dose-ranging. The impact of pre-existing and acquired immunity against the vector and vector stability in several clinically relevant delivery devices was assessed. A hybrid promoter hybrid cytosine guanine dinucleotide (CpG)- free CMV enhancer/elongation factor 1 alpha promoter (hCEF) consisting of the elongation factor 1α promoter and the cytomegalovirus enhancer was most efficacious in both murine lungs and human ALI cultures (both at least 2-log orders above background). The efficacy (at least 14% of airway cells transduced), toxicity and integration site profile supports further progression towards clinical trial and pre-existing and acquired immune responses do not interfere with vector efficacy. The lead rSIV.F/HN candidate expresses functional CFTR and the vector retains 90–100% transduction efficiency in clinically relevant delivery devices. The data support the progression of the F/HN-pseudotyped lentiviral vector into a first-in-man CF trial in 2017.