A project of clinical application in a gene therapy for chronic granulomatous disease with gp91-phox deficiency

自1995年以来，A. Fischer的X-SCID患者基因治疗方案。首先分析了200多例慢性肉芽肿病（CGD）患者，并分析了具有gp 91-phox缺陷的剪接突变的患者中IFN-γ应答的遗传机制（血98：436-441，2001）。在我们与Sugimoto博士合作的3年项目中，我们构建了双顺反子逆转录病毒载体Ha-MDR-IRES-gp 91，MDR 1和gp 91的共表达，gp 91是导致X连锁慢性肉芽肿病（X-CGD）的基因。来自X-CGD患者的EB病毒转化的B细胞和来自用Ha-MDR-IRES-gp 91转导的X-CGD患者的人CD 34阳性细胞共表达P-糖蛋白和gp 91，并且获得了产生超氧化物的活性。移植Ha-MDR-IRES-gp 91转导的骨髓细胞的小鼠显示P-gl共表达， ...更多信息 外周血单个核细胞中的糖蛋白和gp 91。紫杉醇反复给药后，P-糖蛋白和gp 91阳性细胞的比例维持了1年以上。这些结果表明，人多药耐药基因（MDR 1）与治疗基因的共表达为遗传修饰的细胞提供了可选择的生长优势（J Gene Med vol.4，2003）。与Sumimoto博士合作，我们阐明了第四种胞浆因子p40（phox）通过调节p67（phox）和p47（phox）的膜募集参与吞噬细胞氧化酶的激活通过PB 1-PC与p67（phox）的相互作用。(EMBO J 2002 Dec 2 ; 21（23）：6312-20）。此外，我们克隆了与p47 phox和p67 phox同源的新蛋白质的人cDNA，分别命名为p41 nox和p51 nox。基于组合研究，我们发现新同系物p41 nox和p51 nox可能一起或与经典同系物组合起作用，从而激活两种Nox家族氧化酶（J Biol Chem 2003出版中）。少

Clinical gene therapy was required more fundamental and clinical evidences for safety since 3-lymopoproliferative disease had been reported in A. Fischer's gene therapy protocol for X-SCID patients. More than 200 patients with chronic granulomatous disease (CGD) were first analyzed and a genetic mechanisms of IFN-gamma responsibility was analyzed in the patients with the splice mutation of gp91-phox deficiency (Blood 98 : 436-441, 2001).In our 3-year's project, collaborating with Dr. Sugimoto, we constructed a bicistronic retrovirus vector, Ha-MDR-IRES-gp91, for the co-expression of MDR1 and gp91, a gene responsible for X-linked chronic granulomatous disease (X-CGD). Epstein-Barr virus-transformed B cells from X-CGD patients and human CD34-positive cells from an X-CGD patient transduced with Ha-MDR-IRES-gp91 co-expressed P-glycoprotein and gp91, and acquired superoxide-generating activity. Mice transplanted with Ha-MDR-IRES-gp91-transduced bone marrow cells showed co-expression of P-gl … More ycoprotein and gp91 in peripheral blood mononuclear cells. After repeatedly administration of paclitaxel, the ratio of P-glycoprotein- and gp91-positive cells were maintained for over 1 year. These results suggest that co-expression of a human multidrug resistance gene (MDR1) with a therapeutic gene affords selectable growth advantage to genetically modified cells (J Gene Med vol. 4, 2003).In a collaboration with Dr. Sumimoto, we clarified the fourth cytosolic factor p40 (phox) participates in activation of the phagocyte oxidase by regulating membrane recruitment of p67 (phox) and p47 (phox) via the PB1-PC interaction with p67 (phox). (EMBO J 2002 Dec 2 ; 21(23) : 6312-20). In addition, we cloned human cDNAs for novel proteins homologous to p47phox and p67phox, designated p41nox and p51nox, respectively. Based on the combination studies, we found that the novel homologues p41nox and p51nox likely function together or in combination with a classical one, thereby activating the two Nox family oxidases (J Biol Chem 2003 in press). Less