Therapeutic angiogenesis using noble human VEGF-E chimeric genes and autologous bone marrow mononuclear cells

使用高贵的人 VEGF-E 嵌合基因和自体骨髓单核细胞进行治疗性血管生成

• 批准号: 16390221
• 负责人: MUROHARA Toyoaki
• 金额: $ 9.15万
• 依托单位: Nagoya University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2004
• 资助国家: 日本
• 起止时间: 2004 至 2005
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-16390221/
• 关键词: Therapeutic angiogenesis; Vascular regeneration; Regenerative medicine; Gene therapy; Growth factor; Bone marrow cells; Stem cells; Endothelial progenitor cells; 血管再生; 血管成長因子; 再生医療; 虚血性心疾患; 血管透過性; 炎症反応

BackgroundVascular endothelial growth factor-A (VEGF-A) promotes angiogenesis but causes adverse side effects such as edema or tissue inflammation. VEGF-E, found in the genome of the Orf virus, specifically binds to VEGF receptor-2 and shows mitotic activity on endothelial cells. Recently, we created two forms of VEGF-E and human placental growth factor (P1GF) chimera genes (VEGF-E chimera #9 and VEGF-E chimera #33), which are humanized genes holding VEGF-E function but less antigenicity.Methods and ResultsWe examined potential proangiogenic activities of these chimera genes. Four types of expression plasmids (pCDNA3.1-LacZ, phVEGF-A, pVEGF-Echimera#9 and pVEGF-Echimera#33, n=8 each) were administered in a rat model of hindlimb ischemia. Either pVEGF-E chimera #9, pVEGF-E chimera #33 or phVEGF-A significantly increased the ratio of ischemic/normal hindlimb blood-flow compared to the control pCDNA3.1-LacZ treated group. Histochemical staining by alkaline phosphatase also revealed that either pVEGF-Echimera#9, pVEGF-Echimera#33 or phVEGF-A increased the capillary density compared to the pCDNA3.1-LacZ treated group. Furthermore, immunostaining for anti-ED1 revealed that lower number of macrophages were infiltrated in both pVEGF-Echimera#9 and pVEGF-Echimera#33 groups compared to the phVEGF-A group.ConclusionsNovel VEGF-E/human P1GF chimera genes, pVEGF-Echimera#9 and pVEGF-Echimera#33, significantly stimulated angiogenesis in response to tissue ischemia, whose extent was almost identical to that induced by phVEGF-A with less tissue inflammation responses.

背景血管内皮生长因子-A（VEGF-A）促进血管生成，但也引起水肿或组织炎症等不良副作用。VEGF-E，发现于Orf病毒的基因组中，特异性结合VEGF受体-2，并在内皮细胞上显示有丝分裂活性。最近，我们创建了两种形式的VEGF-E和人胎盘生长因子（P1 GF）嵌合体基因（VEGF-E嵌合体#9和VEGF-E嵌合体#33），这是人源化的基因持有VEGF-E功能，但抗原性较低。将四种类型的表达质粒（pCDNA3.1-LacZ、phVEGF-A、pVEGF-Echimera#9和pVEGF-Echimera#33，每种n=8）给予大鼠后肢缺血模型。与对照pCDNA3.1-LacZ处理组相比，pVEGF-E嵌合体#9、pVEGF-E嵌合体#33或phVEGF-A显著增加缺血/正常后肢血流的比率。碱性磷酸酶的组织化学染色也显示，与pCDNA3.1-LacZ处理组相比，pVEGF-Echimera#9、pVEGF-Echimera#33或phVEGF-A增加了毛细血管密度。此外，抗ED 1的免疫染色显示，与phVEGF-A组相比，在pVEGF-Echimera #9和pVEGF-Echimera #33组中浸润的巨噬细胞的数量较低。其程度与phVEGF-A诱导的几乎相同，具有较少的组织炎症反应。

项目成果

Combination therapy using angiopoietin-1 plasmid gene and autologous bone marrow cell implantation promotes functional anigiogenesis.

使用血管生成素-1质粒基因和自体骨髓细胞植入的联合治疗可促进功能性血管生成。

• 发表时间: 2006
• 期刊: Arterioscler.Thromb.Vasc.Biol. (in press)
• 作者: Kobayashi K;Kondo T;Inoue N;Aoki M;Mizuno M;Komori K;Yoshida J;Murohara T.
• 通讯作者: Murohara T.

Catheter-based prostacyclin synthase gene transfer prevents in-stent restenosis in rabbit atheromatous arteries.

基于导管的前列环素合酶基因转移可预防兔动脉粥样硬化的支架内再狭窄。

• 发表时间: 2004
• 期刊: Cardiovascular Research 61
• 作者: Numaguchi;Murohara et al.
• 通讯作者: Murohara et al.

Autologous bone-marrow mononuclear cell implantation improves endothelium-dependent vasodilation in patients with limb ischemia

• DOI: 10.1161/01.cir.0000121427.53291.78
• 发表时间: 2004-03-16
• 期刊: CIRCULATION
• 影响因子: 37.8
• 作者: Higashi, Y;Kimura, M;Yoshizumi, M
• 通讯作者: Yoshizumi, M

Augmentation of therapeutic angiogenesis using genetically-modified human endothelial progenitor cells with altered glycogen synthase kinase-3b activity.

使用糖原合成酶激酶 3b 活性改变的转基因人内皮祖细胞增强治疗性血管生成。

• 发表时间: 2004
• 期刊: Journal of Biological Chemistry 279
• 作者: Choi;Murohara et al.
• 通讯作者: Murohara et al.

Combination therapy using angiopoietin-1 plasmid gene and autologous bone marrow cell implantation promotes functional angiogenesis.

使用 angiopoietin-1 质粒基因和自体骨髓细胞植入的联合治疗可促进功能性血管生成。

• 发表时间: 2006
• 期刊: Arterioscler. Thromb. Vasc. Biol (In press)
• 作者: Kobayashi K;Kondo T;Inoue N;Aoki M;Mizuno M;Komori K;Yoshida J;Murohara T
• 通讯作者: Murohara T