Ontario Cancer Institute, University Health Network, Toronto, M5G 2M1, Canada.
Hum Gene Ther. 2011 Jun;22(6):679-87. doi: 10.1089/hum.2010.195. Epub 2011 Mar 25.
Farber disease is a rare lysosomal storage disorder (LSD) that manifests due to acid ceramidase (AC) deficiencies and ceramide accumulation. We present a preclinical gene therapy study for Farber disease employing a lentiviral vector (LV-huAC/huCD25) in three enzymatically normal nonhuman primates. Autologous, mobilized peripheral blood (PB) cells were transduced and infused into fully myelo-ablated recipients with tracking for at least 1 year. Outcomes were assessed by measuring the AC specific activity, ceramide levels, vector persistence/integration, and safety parameters. We observed no hematological, biochemical, radiological, or pathological abnormalities. Hematological recovery occurred by approximately 3 weeks. Vector persistence was observed in PB and bone marrow (BM) cells by qualitative and quantitative PCR. We did not observe any clonal proliferation of PB and BM cells. Importantly, AC-specific activity was detected above normal levels in PB and BM cells analyzed post-transplantation and in spleens and livers at the endpoint of the study. Decreases of ceramide in PB cells as well as in spleen and liver tissues were seen. We expect that this study will provide a roadmap for implementation of clinical gene therapy protocols targeting hematopoietic cells for Farber disease and other LSDs.
法伯病是一种罕见的溶酶体贮积症(LSD),由于酸性鞘脂酶(AC)缺乏和神经酰胺积累而发病。我们在 3 只酶正常的非人类灵长类动物中进行了法伯病的临床前基因治疗研究,采用了慢病毒载体(LV-huAC/huCD25)。对动员后的自体外周血(PB)细胞进行转导,并输注给完全骨髓清除的受者,至少随访 1 年。通过测量 AC 特异性活性、神经酰胺水平、载体持续性/整合以及安全性参数来评估结果。我们未观察到任何血液学、生化学、影像学或病理学异常。大约 3 周后出现血液学恢复。通过定性和定量 PCR 观察到 PB 和骨髓(BM)细胞中存在载体持续性。我们未观察到 PB 和 BM 细胞的任何克隆性增殖。重要的是,我们在移植后和研究终点时的脾脏和肝脏组织中检测到 PB 和 BM 细胞中 AC 特异性活性高于正常水平。我们还观察到 PB 细胞、脾脏和肝脏组织中神经酰胺的减少。我们期望本研究将为针对法伯病和其他 LSD 靶向造血细胞的临床基因治疗方案的实施提供蓝图。