Pastore Nunzia, Nusco Edoardo, Piccolo Pasquale, Castaldo Sigismondo, Vaníkova Jana, Vetrini Francesco, Palmer Donna J, Vitek Libor, Ng Philip, Brunetti-Pierri Nicola
1 Telethon Institute of Genetics and Medicine , Naples 80131, Italy .
Hum Gene Ther Methods. 2013 Oct;24(5):321-7. doi: 10.1089/hgtb.2013.108.
Crigler-Najjar syndrome type I is caused by mutations of the uridine diphospho-glucuronosyl transferase 1A1 (UGT1A1) gene resulting in life-threatening increase of serum bilirubin. Life-long correction of hyperbilirubinemia was previously shown with intravenous injection of high doses of a helper-dependent adenoviral (HDAd) vector expressing UGT1A1 in the Gunn rat, the animal model of Crigler-Najjar syndrome. However, such high vector doses can activate an acute and potentially lethal inflammatory response with elevated serum interleukin-6 (IL-6). To overcome this obstacle, we investigated safety and efficacy of direct injections of low HDAd doses delivered directly into the liver parenchyma of Gunn rats. Direct hepatic injections performed by either laparotomy or ultrasound-guided percutaneous injections were compared with the same doses given by intravenous injections. A greater reduction of hyperbilirubinemia and increased conjugated bilirubin in bile were achieved with 1 × 10(11) vp/kg by direct liver injections compared with intravenous injections. In sharp contrast to intravenous injections, direct hepatic injections neither raised serum IL-6 nor resulted in thrombocytopenia. In conclusion, ultrasound-guided percutaneous injection of HDAd vectors into liver parenchyma resulted in improved hepatocyte transduction and reduced toxicity compared with systemic injections and is clinically attractive for liver-directed gene therapy of Crigler-Najjar syndrome.
Ⅰ型克里格勒-纳贾尔综合征是由尿苷二磷酸葡萄糖醛酸基转移酶1A1(UGT1A1)基因突变引起的,可导致血清胆红素危及生命的升高。先前在克里格勒-纳贾尔综合征的动物模型冈恩大鼠中,通过静脉注射高剂量表达UGT1A1的辅助依赖型腺病毒(HDAd)载体,可实现高胆红素血症的终身纠正。然而,如此高的载体剂量可激活急性且可能致命的炎症反应,导致血清白细胞介素-6(IL-6)升高。为克服这一障碍,我们研究了直接向冈恩大鼠肝实质内注射低剂量HDAd的安全性和有效性。将通过剖腹手术或超声引导下经皮注射进行的直接肝内注射与静脉注射相同剂量进行比较。与静脉注射相比,通过直接肝内注射给予1×10¹¹ vp/kg剂量可使高胆红素血症得到更大程度的降低,胆汁中结合胆红素增加。与静脉注射形成鲜明对比的是,直接肝内注射既不会升高血清IL-6,也不会导致血小板减少。总之,与全身注射相比,超声引导下经皮将HDAd载体注射到肝实质可改善肝细胞转导并降低毒性,对Ⅰ型克里格勒-纳贾尔综合征的肝脏定向基因治疗具有临床吸引力。