AAV-mediated factor IX gene transfer to skeletal muscle in patients with severe hemophilia B
AAV-mediated factor IX gene transfer to skeletal muscle in patients with severe hemophilia B
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DOI：
10.1182/blood-2002-10-3296
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发表时间：
2003-04-15
期刊：
影响因子：
20.3
通讯作者：
Glader, B
中科院分区：
文献类型：
--
作者：
Manno, CS;Chew, AJ;Glader, B
血友病B是一种由功能性凝血因子IX(F.IX)缺失引起的X连锁凝血病。此前，我们通过肌肉注射表达F.IX的重组腺相关病毒(RAAV)载体，为基因转移作为一种治疗小鼠和血友病狗疾病的方法建立了实验基础。在这项研究中，我们研究了这种方法在血友病B患者中的安全性。在一项开放标记的剂量-递增研究中，由于错义突变导致的严重血友病6(F.IX<；110 6)成年男性被注射到多个肌肉内部位注射In rAAV载体。在2×10(11)载体基因组(VG)/公斤到1.8×10(12)VG/公斤的剂量范围内，没有证据表明局部或全身毒性--直到注射后40个月。注射部位肌肉活检在载体注射后2-10个月进行，Southern印迹和免疫组织化学染色证实转基因表达。预先存在的。甲型肝炎病毒的高滴度抗体不能阻止基因转移或表达。尽管有强有力的证据表明存在基因转移和表达，但在所有病例中，循环中的F.IX水平都低于2%，大多数低于1%。尽管需要更广泛的肌肉纤维转导才能开发出一种在所有受试者中可靠地将循环水平提高到1%以上的疗法，但首次非肠道注射rAAV的这些结果表明，在所测试的剂量下，通过肌肉途径给药AAV载体是安全的，并以与动物相似的方式影响人类的基因转移和表达。(C)2003年，由美国血液病学会提供。
Hemophilia B is an X-linked coagulopathy caused by absence of functional coagulation factor IX (F.IX). previously, we established an experimental basis for gene transfer as a method of treating the disease in mice and hemophilic dogs through intramuscular injection of a recombinant adeno-associated viral (rAAV) vector expressing F.IX. In this study we, investigated the safety of this approach in patients with hemophilia B. In an open-label dose-escalation study, adult men with severe hemophilia 6 (F.IX < 110 6) due to a missense mutation were injected at multiple intramuscular sites with in rAAV vector. At doses ranging from 2 x 10(11) vector genomes (vg)/kg to, 1.8 X 10(12) vg/kg, there was no evidence-of local or systemic toxicity up to 40: months after injection. Muscle biopsies of injection sites performed 2 to 10 months after vector administration I confirmed gene transfer as evidenced by, Southern blot and transgene expression as evidenced by immunohistochemical staining. Preexisting. high-titer antibodies to AAV did not prevent gene transfer or expression. Despite strong evidence for gene transfer and expression, circulating levels of F.IX were in all cases less than 2% and most were less than 1%. Although more extensive transduction of muscle fibers will be required to develop a therapy that reliably raises circulating levels to moire than 1% in all subjects, these results of the first parenteral administration of rAAV demonstrate that administration of AAV vector by the intramuscular route is safe at the doses tested and effects gene transfer and expression in humans in a manner similar to that seen in animals. (C) 2003 by The American Society of Hematology.