AAV vector delivery to skeletal muscle, platform for therapeutic protein delivery

综合征(SIDS)或合并心脏和骨骼肌病。这些疾病的治疗主要包括饮食控制，到目前为止还远远不是最理想的。近年来，用于高效转导肝细胞和肌纤维的重组腺相关病毒载体的发展为粮农组织疾病的研究提供了新的工具。具体地说，我们的实验室已经制备了表达FAO酶的rAAV载体，这些酶缺乏会导致肌病，如短链酰基辅酶A脱氢酶(SCAD)和长链酰基辅酶A脱氢酶(LCAD)。来自这些酶缺陷患者的人类细胞系是可用的，这两种疾病的突变小鼠模型都存在。我们建议利用表达粮农组织酶的rAAV载体，试图揭示粮农组织疾病的病理生物学，并更好地确定这些疾病的分子或细胞治疗的终点。这将通过三个具体目标来实现：(1)评估细胞群体或器官中有限百分比的SCAD缺陷或LCAD缺陷细胞的遗传校正对脂肪酸氧化的生化校正的影响程度。(2)确定受体结合和进入是否是限制rAAV在完整的哺乳动物肝脏或肌束中稳定转导的步骤。(3)验证LCAD和VLCAD缺陷的肝脏病理是肝细胞内有毒代谢物的蓄积而不是初级能量衰竭的假设。(4)确定在宫内注射或新生儿静脉注射后广泛传播媒介后，全球纠正小鼠FAO缺乏症的表型是否更有效。从这些研究中获得的信息也可以用来指导其他器官导向疗法的可行性，可能包括干细胞移植。

Syndrome (SIDS) or with a combined cardiac and skeletal myopathy. Treatment of these disorders has consisted primarily of dietary manipulation and has been far less than optimal to this point. The recent development of recombinant adeno-associated virus (rAAV) vectors for highly efficient transduction of hepatocytes and myofibers present new tools for the study of FAO disorders. Specifically, our laboratory has produced rAAV vectors expressing FAO enzymes whose deficiency results in myopathy, such as short-chain acyl CoA dehydrogenase (SCAD) and long-chain acyl CoA dehydrogenase (LCAD). Human cell lines from patients deficient in these enzymes are available, and mutant mouse models exist for both of these disorders. We propose to utilize rAAV vectors expressing FAO enzymes in an attempt to unravel the pathobiology of FAO disorders and to better define endpoints for molecular or cell-based therapies of these disorders. This will be accomplished in three specific aims: (1) To assess the extent to which genetic correction of a limited percentage of SCAD deficient or LCAD deficient cells within a cell population or organ can affect biochemical correction of fatty acid oxidation. (2) To determine whether receptor binding and entry are limiting steps for stable transduction by rAAV in a intact mammalian liver or muscle bundle. (3) To test the hypothesis that the liver pathology observed in LCAD and VLCAD deficiencies are secondary to the accumulation of toxic metabolites as opposed to primary energy failure within hepatocytes. (4) To determine whether global phenotypic correction of FAO deficiency in mice is more effective after widespread vector delivery after intrauterine or neonatal IV injection. The information gained from these studies could also be used to guide the feasibility of other organ-directed therapies, potentially including stem cell transplantation.