Lentivirus Mediated Correction of Artemis-Deficient Severe Combined Immunodeficiency.
Lentivirus Mediated Correction of Artemis-Deficient Severe Combined Immunodeficiency.
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慢病毒介导纠正 Artemis 缺陷的严重联合免疫缺陷。
DOI：
10.1089/hum.2016.064
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发表时间：
2017
影响因子：
4.2
通讯作者：
Puck,Jennif
中科院分区：
文献类型：
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作者：
Punwani,Divya;Kawahara,Misako;Yu,Jason;Sanford,Ukina;Roy,Sushmita;Patel,Kiran;Carbonaro,DeniseA;Karlen,AndreaD;Khan,Sara;Cornetta,Kenneth;Rothe,Michael;Schambach,Axel;Kohn,DonaldB;Malech,HarryL;McIvor,RScott;Puck,Jennif
在 B 和 T 淋巴细胞成熟过程中，V(D)J 重组是通过 DNA 双链断裂的产生而启动的。 Artemis 是一种核酸外切酶，对于随后通过非同源末端连接进行修复至关重要。 DCLRE1C（编码 Artemis 的基因）中的突变会导致 T−B−NK+ 严重联合免疫缺陷 (ART-SCID)，并且还会提高对电离辐射和烷化化疗的敏感性。尽管同种异体造血细胞移植可以治疗 ART-SCID，但预处理方案的耐受性较差，导致早期死亡和/或晚期并发症，包括身材矮小、内分泌疾病和牙齿发育不全。然而，如果没有烷化化疗作为预处理，患者通常会出现移植排斥或T细胞有限且B细胞无法恢复。因此，向自体造血干细胞添加正常的DCLRE1CcDNA是治疗ART-SCID的一种有吸引力的策略。我们设计了一种包含人Artemis cDNA、在人内源Artemis启动子（AProArt）转录调控下的自失活慢病毒载体。用 AProArt 慢病毒转导的 ART-SCID 患者的成纤维细胞显示放射敏感性得到纠正。来自 ART-SCID 患者的动员外周血 CD34+ 细胞以及来自 Artemis 缺陷小鼠的造血干细胞显示，在 AProArt 转导后，T 细胞和 B 细胞发育得到恢复。在体外永生化测定中，用 AProArt 转导的小鼠造血细胞没有表现出重新接种潜力的增加，并且对 AProArt 慢病毒插入的分析表明对可以激活癌基因的位点没有偏好。这些功效和安全性研究结果支持开展 ART-SCID 基因添加疗法临床试验。
During B and T lymphocyte maturation, V(D)J recombination is initiated by creation of DNA double-strand breaks. Artemis is an exonuclease essential for their subsequent repair by nonhomologous end-joining. Mutations inDCLRE1C, the gene encoding Artemis, cause T−B−NK+severe combined immunodeficiency (ART-SCID) and also confer heightened sensitivity to ionizing radiation and alkylating chemotherapy. Although allogeneic hematopoietic cell transplantation can treat ART-SCID, conditioning regimens are poorly tolerated, leading to early mortality and/or late complications, including short stature, endocrinopathies, and dental aplasia. However, without alkylating chemotherapy as preconditioning, patients usually have graft rejection or limited T cell and no B cell recovery. Thus, addition of normalDCLRE1CcDNA to autologous hematopoietic stem cells is an attractive strategy to treat ART-SCID. We designed a self-inactivating lentivirus vector containing human Artemis cDNA under transcriptional regulation of the human endogenous Artemis promoter (AProArt). Fibroblasts from ART-SCID patients transduced with AProArt lentivirus showed correction of radiosensitivity. Mobilized peripheral blood CD34+cells from an ART-SCID patient as well as hematopoietic stem cells from Artemis-deficient mice demonstrated restored T and B cell development following AProArt transduction. Murine hematopoietic cells transduced with AProArt exhibited no increase in replating potential in anin vitroimmortalization assay, and analysis of AProArt lentivirus insertions showed no predilection for sites that could activate oncogenes. These efficacy and safety findings support institution of a clinical trial of gene addition therapy for ART-SCID.