Novel Combination BMP7 and HGF Gene Therapy Instigates Selective Myofibroblast Apoptosis and Reduces Corneal Haze In Vivo.
Novel Combination BMP7 and HGF Gene Therapy Instigates Selective Myofibroblast Apoptosis and Reduces Corneal Haze In Vivo.
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DOI：
10.1167/iovs.17-23308
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发表时间：
2018-02-01
影响因子：
4.4
通讯作者：
Mohan RR
中科院分区：
文献类型：
--
作者：
Gupta S;Fink MK;Ghosh A;Tripathi R;Sinha PR;Sharma A;Hesemann NP;Chaurasia SS;Giuliano EA;Mohan RR
我们使用已建立的兔体内和人体外模型测试了骨形态发生蛋白 7 (BMP7) 和肝细胞生长因子 (HGF) 联合基因疗法治疗预先形成的角膜纤维化的潜力。使用十八只新西兰白兔。碱损伤导致角膜纤维化。疤痕形成后 24 小时，角膜局部接受平衡盐溶液（BSS；n = 6）、聚乙烯亚胺缀合金纳米颗粒（PEI2-GNP）裸质粒（n = 6）或表达 BMP7 和 HGF 基因的 PEI2-GNP 质粒（n = 6）。使用供体人角膜获得原代人角膜成纤维细胞和肌成纤维细胞用于机制研究。通过裂隙灯显微镜、体视显微镜、定量实时PCR、免疫荧光、TUNEL、改良MacDonald-Shadduck评分系统和Draize试验评估基因治疗对角膜纤维化的效果和眼部安全性。 PEI2-GNP 介导的 BMP7+HGF 基因治疗显着降低了活体兔体内的角膜纤维化（BMP7+HGF 治疗组的 Fantes 评分为 0.6，而治疗组为 3.3；P < 0.001）。接受 BMP7+HGF 的角膜显示促纤维化基因的 mRNA 水平显着降低：α-SMA（3.2 倍；P < 0.01）、纤连蛋白（2.3 倍，P < 0.01）、胶原蛋白 I（2.1 倍，P < 0.01）、胶原蛋白 III（1.6 倍，P < 0.01）和胶原蛋白 IV（1.9 倍，P < 0.01）与-治疗角膜相比。此外，与-治疗对照相比，BMP7+HGF治疗的角膜显示肌成纤维细胞显着减少（83%；P < 0.001）。 PEI2-GNP 每微克 DNA 引入 >104 个基因拷贝的 BMP7 和 HGF 基因。重组 HGF 导致角膜肌成纤维细胞凋亡，但不导致成纤维细胞凋亡。局部BMP7+HGF治疗未显示出眼部毒性。局部 BMP7+HGF 基因治疗可治疗角膜纤维化并恢复体内透明度，减轻过度愈合并导致肌成纤维细胞选择性凋亡。
We tested the potential of bone morphogenic protein 7 (BMP7) and hepatocyte growth factor (HGF) combination gene therapy to treat preformed corneal fibrosis using established rabbit in vivo and human in vitro models. Eighteen New Zealand White rabbits were used. Corneal fibrosis was produced by alkali injury. Twenty-four hours after scar formation, cornea received topically either balanced salt solution (BSS; n = 6), polyethylenimine-conjugated gold nanoparticle (PEI2-GNP)-naked plasmid (n = 6) or PEI2-GNP plasmids expressing BMP7 and HGF genes (n = 6). Donor human corneas were used to obtain primary human corneal fibroblasts and myofibroblasts for mechanistic studies. Gene therapy effects on corneal fibrosis and ocular safety were evaluated by slit-lamp microscope, stereo microscopes, quantitative real-time PCR, immunofluorescence, TUNEL, modified MacDonald-Shadduck scoring system, and Draize tests. PEI2-GNP–mediated BMP7+HGF gene therapy significantly decreased corneal fibrosis in live rabbits in vivo (Fantes scale was 0.6 in BMP7+HGF-treated eyes compared to 3.3 in −therapy group; P < 0.001). Corneas that received BMP7+HGF demonstrated significantly reduced mRNA levels of profibrotic genes: α-SMA (3.2-fold; P < 0.01), fibronectin (2.3-fold, P < 0.01), collagen I (2.1-fold, P < 0.01), collagen III (1.6-fold, P < 0.01), and collagen IV (1.9-fold, P < 0.01) compared to the −therapy corneas. Furthermore, BMP7+HGF-treated corneas showed significantly fewer myofibroblasts compared to the −therapy controls (83%; P < 0.001). The PEI2-GNP introduced >104 gene copies per microgram DNA of BMP7 and HGF genes. The recombinant HGF rendered apoptosis in corneal myofibroblasts but not in fibroblasts. Localized topical BMP7+HGF therapy showed no ocular toxicity. Localized topical BMP7+HGF gene therapy treats corneal fibrosis and restores transparency in vivo mitigating excessive healing and rendering selective apoptosis in myofibroblasts.
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影响因子：
17.8
作者：
Ljubimov AV;Saghizadeh M
通讯作者：
Saghizadeh M
影响因子：
1.6
作者：
Donnelly, Kevin S.;Giuliano, Elizabeth A.;Mohan, Rajiv R.
通讯作者：
Mohan, Rajiv R.
影响因子：
6
作者：
Kim, WH;Matsumoto, K;Nakamura, T
通讯作者：
Nakamura, T
影响因子：
3.9
作者：
Foster, A;Resnikoff, S
通讯作者：
Resnikoff, S
DOI：
10.1006/bbrc.1998.8397
发表时间：
1998-04-07
影响因子：
3.1
作者：
Arakaki, N;Kazi, JA;Daikuhara, Y
通讯作者：
Daikuhara, Y