Peptide-conjugated phosphodiamidate oligomer-mediated exon skipping has benefits for cardiac function in mdx and Cmah-/-mdx mouse models of Duchenne muscular dystrophy.
Peptide-conjugated phosphodiamidate oligomer-mediated exon skipping has benefits for cardiac function in mdx and Cmah-/-mdx mouse models of Duchenne muscular dystrophy.
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DOI：
10.1371/journal.pone.0198897
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发表时间：
2018
期刊：
影响因子：
3.7
通讯作者：
Straub VW
中科院分区：
文献类型：
--
作者：
Blain AM;Greally E;McClorey G;Manzano R;Betts CA;Godfrey C;O'Donovan L;Coursindel T;Gait MJ;Wood MJ;MacGowan GA;Straub VW
心衰是杜氏肌营养不良症（DMD）患者死亡的主要原因。反义介导的外显子跳变能够纠正DMD的框外突变，从而产生截断但具有功能的肌营养不良蛋白。然而，传统的反义方法受到心肌吸收不良的限制。在反义分子中加入细胞穿透肽增加了它们的效力，并改善了它们对包括心脏在内的所有肌肉的吸收。我们研究了肽偶联磷酸二酯morpholino寡聚物（P-PMO） Pip6a-PMO在DMD小鼠(mdx小鼠和不太明确的Cmah-/-mdx小鼠（在小鼠Cmah基因中携带类似人类的突变以及DMD突变）中恢复心脏肌营养不良蛋白和功能恢复的功效。在我们的第一项研究中，从12周龄到30周龄，雄性mdx小鼠每两周注射一次Pip6a-PMO，同时模拟注射与年龄匹配的mdx和C57BL10对照。小鼠给药4次，剂量为18 mg/kg，随后给药8次，剂量为12.5 mg/kg。最后一次注射2周后，通过MRI和导尿管分析小鼠的心功能，并收集肌肉用于测定肌营养不良蛋白。在第二项研究中，雄性Cmah-/-mdx小鼠，在8至26周内每两周注射12.5 mg/kg Pip6a-PMO，并在3个时间点（12,18和28周）与模拟注射年龄匹配的mdx， C57BL10和Cmah-/-mdx对照进行MRI评估。小鼠在28周时也接受了MEMRI和导尿管治疗。这使我们能够表征Cmah-/-mdx小鼠的心脏表型，并评估P-PMO对心功能的影响。Pip6a-PMO治疗可显著恢复mdx和Cmah-/-mdx小鼠的肌营养不良蛋白（dystrophin）（分别为37.5%和51.6%），足以显著改善心功能，改善左右心室功能障碍。Cmah-/-mdx小鼠对多巴酚丁胺应激测试表现出异常反应，PIP6a-PMO治疗完全改善了这种情况。这些令人鼓舞的数据表明，完全恢复肌营养不良蛋白可能不需要显着改善DMD患者的心脏预后，并且期望适度水平的肌营养不良蛋白恢复功能改善可能是现实的，这在未来的临床试验中可能是可以实现的。
Cardiac failure is a major cause of mortality in patients with Duchenne muscular dystrophy (DMD). Antisense-mediated exon skipping has the ability to correct out-of-frame mutations in DMD to produce truncated but functional dystrophin. Traditional antisense approaches have however been limited by their poor uptake into cardiac muscle. The addition of cell-penetrating peptides to antisense molecules has increased their potency and improved their uptake into all muscles, including the heart. We have investigated the efficacy of the Peptide-conjugated phosphodiamidate morpholino oligomer (P-PMO) Pip6a-PMO, for restoration of cardiac dystrophin and functional rescue in DMD mice- the mdx mouse and the less well characterised Cmah-/-mdx mouse (which carry a human-like mutation in the mouse Cmah gene as well as a mutation in DMD). In our first study male mdx mice were administered Pip6a-PMO, i.v, fortnightly from 12 to 30 weeks of age alongside mock-injected age-matched mdx and C57BL10 controls. Mice received 4 doses of 18 mg/kg followed by 8 doses of 12.5 mg/kg. The cardiac function of the mice was analysed 2 weeks after their final injection by MRI followed by conductance catheter and their muscles were harvested for dystrophin quantification. In the second study, male Cmah-/-mdx mice, received 12.5 mg/kg Pip6a-PMO, i.v fortnightly from 8 to 26 weeks and assessed by MRI at 3 time points (12, 18 and 28 weeks) alongside mock-injected age-matched mdx, C57BL10 and Cmah-/-mdx controls. The mice also underwent MEMRI and conductance catheter at 28 weeks. This allowed us to characterise the cardiac phenotype of Cmah-/-mdx mice as well as assess the effects of P-PMO on cardiac function. Pip6a-PMO treatment resulted in significant restoration of dystrophin in mdx and Cmah-/-mdx mice (37.5% and 51.6%, respectively), which was sufficient to significantly improve cardiac function, ameliorating both right and left ventricular dysfunction. Cmah-/-mdx mice showed an abnormal response to dobutamine stress test and this was completely ameliorated by PIP6a-PMO treatment. These encouraging data suggest that total restoration of dystrophin may not be required to significantly improve cardiac outcome in DMD patients and that it may be realistic to expect functional improvements with modest levels of dystrophin restoration which may be very achievable in future clinical trials.
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DOI：
10.1186/1532-429x-15-4
发表时间：
2013-01-16
期刊：
Journal of cardiovascular magnetic resonance : official journal of the Society for Cardiovascular Magnetic Resonance
影响因子：
--
作者：
Greally E;Davison BJ;Blain A;Laval S;Blamire A;Straub V;MacGowan GA
通讯作者：
MacGowan GA
影响因子：
10.8
作者：
Jearawiriyapaisarn, Natee;Moulton, Hong M.;Willis, Monte S.
通讯作者：
Willis, Monte S.
影响因子：
64.8
作者：
MONACO, AP;NEVE, RL;KUNKEL, LM
通讯作者：
KUNKEL, LM
影响因子：
10.8
作者：
Bauer, R.;MacGowan, G. A.;Straub, V.
通讯作者：
Straub, V.
影响因子：
168.9
作者：
Cirak, Sebahattin;Arechavala-Gomeza, Virginia;Guglieri, Michela;Feng, Lucy;Torelli, Silvia;Anthony, Karen;Abbs, Stephen;Garralda, Maria Elena;Bourke, John;Wells, Dominic J.;Dickson, George;Wood, Matthew J. A.;Wilton, Steve D.;Straub, Volker;Kole, Ryszard;Shrewsbury, Stephen B.;Sewry, Caroline;Morgan, Jennifer E.;Bushby, Kate;Muntoni, Francesco
通讯作者：
Muntoni, Francesco