From the Department of Genetics, Yale School of Medicine, New Haven (A.L., K.M., S.H., K.W., S. Pajusalu, M.L.), and Cure Rare Disease, Woodbridge (R.H.) - both in Connecticut; the Departments of Pediatrics (B.W., A.K., R.A., D.K., T.F.) and Neurology (A.R.B.) and Horae Gene Therapy Center and the Li Weibo Institute for Rare Diseases Research (A.K., M.B., K. Sylvia, A.R.B., R.A., D.K., S. Parajuli, T.G., T.F.), University of Massachusetts Chan Medical School, Worcester, the Department of Pathology (J.P., C.K.C., H.L.), the Division of Genetics (J.M.S.), and Department of Cardiology (J.L.), Boston Children's Hospital, and Harvard Medical School (J.P., C.K.C., H.L.), Boston, and Charles River Laboratories, Wilmington (L.B., K. Sutton) - all in Massachusetts; the Department of Clinical Genetics, Institute of Clinical Medicine, University of Tartu (S. Pajusalu), and the Genetics and Personalized Medicine Clinic, Tartu University Hospital (S. Pajusalu) - both in Tartu, Estonia; and Regulatory Innovation, Raleigh, NC (D.B.).
N Engl J Med. 2023 Sep 28;389(13):1203-1210. doi: 10.1056/NEJMoa2307798.
We treated a 27-year-old patient with Duchenne's muscular dystrophy (DMD) with recombinant adeno-associated virus (rAAV) serotype 9 containing dCas9 (i.e., "dead" Cas9, in which the Cas9 nuclease activity has been inactivated) fused to VP64; this transgene was designed to up-regulate cortical dystrophin as a custom CRISPR-transactivator therapy. The dose of rAAV used was 1×10 vector genomes per kilogram of body weight. Mild cardiac dysfunction and pericardial effusion developed, followed by acute respiratory distress syndrome (ARDS) and cardiac arrest 6 days after transgene treatment; the patient died 2 days later. A postmortem examination showed severe diffuse alveolar damage. Expression of transgene in the liver was minimal, and there was no evidence of AAV serotype 9 antibodies or effector T-cell reactivity in the organs. These findings indicate that an innate immune reaction caused ARDS in a patient with advanced DMD treated with high-dose rAAV gene therapy. (Funded by Cure Rare Disease.).
我们用携带 dCas9（即“失活” Cas9，其 Cas9 核酸内切酶活性已被失活）融合 VP64 的重组腺相关病毒（rAAV）血清型 9 治疗了一名 27 岁的杜氏肌营养不良症（DMD）患者；这种转基因设计用于上调皮质肌营养不良蛋白，作为一种定制的 CRISPR 转录激活治疗。使用的 rAAV 剂量为每千克体重 1×10 个载体基因组。转基因治疗后 6 天出现轻度心功能障碍和心包积液，随后出现急性呼吸窘迫综合征（ARDS）和心脏骤停；患者两天后死亡。尸检显示严重弥漫性肺泡损伤。肝脏中转基因的表达很少，器官中也没有 AAV 血清型 9 抗体或效应 T 细胞反应的证据。这些发现表明，一名接受高剂量 rAAV 基因治疗的晚期 DMD 患者发生 ARDS 的原因是固有免疫反应。（由 Cure Rare Disease. 资助）。
