Differential proteomics and functional research following gene therapy in a mouse model of Leber congenital amaurosis.
Differential proteomics and functional research following gene therapy in a mouse model of Leber congenital amaurosis.
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DOI：
10.1371/journal.pone.0044855
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发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Li W
中科院分区：
文献类型：
--
作者：
Zheng Q;Ren Y;Tzekov R;Zhang Y;Chen B;Hou J;Zhao C;Zhu J;Zhang Y;Dai X;Ma S;Li J;Pang J;Qu J;Li W
莱伯先天性黑蒙 (LCA) 是遗传性视网膜变性最严重的形式之一，可由至少 15 个不同基因的突变引起。为了阐明 LCA 眼的蛋白质组差异，一组视网膜变性 12 (rd12) 小鼠（一种由 RPE65 基因突变引起的 LCA2 模型）在一只眼中视网膜下注射 AAV 载体 (scAAV5-smCBA-hRPE65)，而对侧眼作为对照。在 P14 和 P21 上比较未处理的 rd12 和正常对照视网膜之间的蛋白质组学，以及在 P42 上比较处理和未处理的 rd12 视网膜和对照视网膜之间的蛋白质组学。视网膜电图 (ERG) 证实，rd12 小鼠的基因治疗恢复了治疗眼睛的视网膜功能。蛋白质组学分析成功鉴定出 3 组中 39 种表达差异的蛋白质。进一步研究了参与细胞凋亡和神经保护调节的 3 种蛋白（α A 晶状体蛋白、热休克蛋白 70 和过氧化还原蛋白 6）的表达。免疫荧光、蛋白质印迹和实时PCR证实了它们表达的定量变化。此外，细胞培养研究表明过氧化还原蛋白 6 可以在 rd12 小鼠中发挥抗氧化作用。我们的研究结果支持基因治疗在 LCA2 患者中的可行性，并支持 α A 晶状体蛋白、热休克蛋白 70 和过氧化还原蛋白 6 在涉及 LCA2 疾病过程的发病机制中的作用。
Leber congenital amaurosis (LCA) is one of the most severe forms of inherited retinal degeneration and can be caused by mutations in at least 15 different genes. To clarify the proteomic differences in LCA eyes, a cohort of retinal degeneration 12 (rd12) mice, an LCA2 model caused by a mutation in the RPE65 gene, were injected subretinally with an AAV vector (scAAV5-smCBA-hRPE65) in one eye, while the contralateral eye served as a control. Proteomics were compared between untreated rd12 and normal control retinas on P14 and P21, and among treated and untreated rd12 retinas and control retinas on P42. Gene therapy in rd12 mice restored retinal function in treated eyes, which was demonstrated by electroretinography (ERG). Proteomic analysis successfully identified 39 proteins expressed differently among the 3 groups. The expression of 3 proteins involved in regulation of apoptosis and neuroptotection (alpha A crystallin, heat shock protein 70 and peroxiredoxin 6) were investigated further. Immunofluorescence, Western blot and real-time PCR confirmed the quantitative changes in their expression. Furthermore, cell culture studies suggested that peroxiredoxin 6 could act in an antioxidant role in rd12 mice. Our findings support the feasibility of gene therapy in LCA2 patients and support a role for alpha A crystallin, heat shock protein 70 and peroxiredoxin 6 in the pathogenetic mechanisms involved in LCA2 disease process.
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影响因子：
--
作者：
Jacobson, Samuel G.;Cideciyan, Artur V.;Ratnakaram, Ramakrishna;Heon, Elise;Schwartz, Sharon B.;Roman, Alejandro J.;Peden, Marc C.;Aleman, Tomas S.;Boye, Sanford L.;Sumaroka, Alexander;Conlon, Thomas J.;Calcedo, Roberto;Pang, Ji-Jing;Erger, Kirsten E.;Olivares, Melani B.;Mullins, Cristina L.;Swider, Malgorzata;Kaushal, Shalesh;Feuer, William J.;Iannaccone, Alessandro;Fishman, Gerald A.;Stone, Edwin M.;Byrne, Barry J.;Hauswirth, William W.
通讯作者：
Hauswirth, William W.
影响因子：
2
作者：
DeLaPaz, MA;Zhang, J;Fridovich, I
通讯作者：
Fridovich, I
影响因子：
56.9
作者：
BECKMANN, RP;MIZZEN, LA;WELCH, WJ
通讯作者：
WELCH, WJ
影响因子：
4.2
作者：
Jacobson, Samuel G.;Boye, Sanford L.;Hauswirth, William W.
通讯作者：
Hauswirth, William W.
影响因子：
3.4
作者：
Fort, Patrice E.;Lampi, Kirsten J.
通讯作者：
Lampi, Kirsten J.