Duchenne型肌营养不良临床诊治的系统性研究
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摘要
第一部分西南地区假肥大型肌营养不良的诊治现状及生存质量分析
目的：分析西南地区假肥大型肌营养不良（Duchenne/Beckermuscular dystrophy, DMD/BMD）患儿的诊治现状及生存质量（qualityof life, QoL）状况，以提高其诊疗水平。
方法：选取2009年12月～2011年12月我院疑诊DMD/BMD病例294例，采用多重聚合酶链反应（multiplepolymerase chain reaction, mPCR）+短串联重复序列（short tandemrepeat, STR）多态性连锁分析法行dystrophin基因检测。随机抽取17例基因检测阴性患儿行腓肠肌活检，光镜下观察病理改变。遵循量表引进程序，首次将儿童生存质量量表PedsQLTM3.0神经肌肉疾病模块（pediatric quality of life inventoryTM3.0neuromuscular module,PedsQLTM3.0NMM）翻译、修订并形成PedsQLTM3.0NMM中文版后，随机抽取39例确诊DMD/BMD患儿及其家长，调查其QoL。并随访治疗情况。
结果：294例疑诊病例mPCR+STR法检出外显子/STR缺失的阳性率为32.65%。外显子48、50和STR49CA、50CA缺失率最高。17例活检的阳性率为82.35%。PedsQLTM3.0NMM中文版的缺失率<1%，克伦巴赫系数α（Cronbach’s α）>0.7，评定者间的信度（intraclasscorrelation coefficient, ICC）>0.6，各条目与所属维度的Spearman相关系数（correlation coefficient of Spearman, rs）呈中、高度相关，相对拟合指数（Comparative fit index, CFI）>0.9。DMD/BMD患儿病情越重，疾病维度评分越低，差异有显著性（P <0.05）；而交流、家庭维度评分普遍较低，差异无统计学意义（P>0.05）。101例确诊的DMD患儿中，17.82%予口服泼尼松0.75mg/kg/d治疗，大部分未予治疗，观察随访中。
结论：mPCR+STR法可提高dystrophin基因检测的准确性，但阳性率不高；肌肉活检是确诊手段的良好补充。PedsQLTM3.0NMM中文版具有较好的信度和效度，可应用于我国DMD/BMD患儿QoL的评价。病情轻重是影响DMD/BMD患儿QoL的主要因素；他们普遍缺乏良好的交流，家庭经济条件较差。大部分DMD患儿未予治疗。制定适合我国国情的DMD/BMD临床诊治指南尤为迫切。
第二部分糖皮质激素治疗Duchenne型肌营养不良的临床疗效观察
目的：参考国外Duchenne型肌营养不良（Duchenne musculardystrophy, DMD）的临床诊治指南，多学科协作，进行糖皮质激素（glucocorticoid，GC）治疗DMD的前瞻性、随机、对照的临床试验，系统地评估GC治疗DMD的疗效和远期预后的影响，以期为西南地区DMD的早期诊断、早期干预、疗效评价及副反应监测提供建设性的指导意见。
方法：选取2010年12月～2012年12月我院经基因或病理检测确诊为DMD患儿66例，按随机数字表法随机抽取进入泼尼松组（36例）和对照组（30例）。为期6～12月泼尼松0.75mg/kg/d治疗的前瞻性临床试验后，比较治疗前后DMD患儿的肌力、肌肉功能、肌酸激酶（creatine kinase, CK）值、骨骼肌B超、生存质量（qualityof life, QoL）的变化，并监测药物副反应。
结果：治疗6～12月后，泼尼松组患儿的肌力、肌肉功能基本保持稳定并有好转，与治疗前比较，差异无显著性（P>0.05）。骨骼肌B超回声减弱，皮下脂肪厚度降低；QoL明显改善，与治疗前及同时点对照组比较，差异有显著性（P <0.05）。治疗6月后，泼尼松组患儿的CK值明显增高，与治疗前及同时点对照组比较，差异有显著性（P <0.05）；此后CK值逐渐下降，但尚未恢复正常范围。对照组患儿随病情的进展，肌力、肌肉功能下降明显，骨骼肌B超回声及QoL有明显恶化趋势，CK值有逐渐下降趋势，与治疗前比较，差异有显著性（P <0.05）。临床试验过程中，大部分DMD患儿无明显副反应，治疗前后，两组DMD患儿的体重、身高、血压（舒张压）相比较，差异均无显著性（P>0.05）。
结论：泼尼松0.75mg/kg/d适合西南地区DMD患儿的中长期治疗，副反应少。DMD患儿血清CK值的变化具有诊断价值，但存在个体差异，且受多种因素的影响，无法作为评估药物疗效的客观指标。骨骼肌B超不但能判断肌肉病变部位、范围、程度，为疾病的早期诊断、鉴别诊断及病情随访提供客观依据；而且动态监测骨骼肌B超的变化可作为评估DMD药物疗效的客观指标。这些结果可初步为西南地区DMD的早期诊断、早期干预、疗效评价及副反应监测提供建设性的指导意见。
第三部分肌肉特异性microRNAs在Duchenne型肌营养不良中的表达及意义
目的：通过检测Duchenne型肌营养不良（Duchenne musculardystrophy, DMD）血清肌肉特异性microRNAs（miRNAs）的表达水平，探讨其与DMD病情严重程度的相关性，以期作为DMD诊断的新型生物标志物。
方法：2010年12月～2012年12月我院经基因或病理检测确诊的DMD患儿66例，按随机数字表法随机抽取进入实验组（39例）。于同一时期本院儿童保健科健康体检的健康男性儿童中抽取对照组36例。行肌力、肌肉功能、生存质量（quality of life, QoL）评估及血清肌酸激酶（creatine kinase, CK）检测后，采用实时定量荧光聚合酶链反应（Real-time quantitative polymerase chain reaction,RT-qPCR）法检测血清肌肉特异性miRNAs的表达水平，并与血清CK值及肌力、肌肉功能和QoL进行比较，探讨肌肉特异性miRNAs与DMD病情严重程度的相关性。
结果：实验组血清肌肉特异性miRNAs的表达水平较对照组明显增高，差异有显著性（P <0.01）。肌肉特异性miRNAs与年龄之间无相关性（P>0.05），与肌力、肌肉功能及QoL之间存在显著的负相关性（P <0.05），尤以miRNA-206为显著。CK值与年龄之间存在显著的相关性（P <0.05），与肌力、肌肉功能及QoL无相关性（P>0.05）。受试者工作特征（receiver operating characteristic, ROC）曲线分析显示，肌肉特异性miRNAs的AUC≥0.9，诊断价值较高，灵敏度和特异度好，尤以miRNA-206为著。
结论：肌肉特异性miRNAs在DMD患儿的血清中高度表达，稳定性强，并与病情的严重程度密切相关，可作为DMD诊断的新型生物标志物，对指导相关基础与临床研究具有重要的意义。
PART I DUCHENNE/BECKER MUSCULARDYSTROPHY IN SOUTHWEST OF CHINA
Objective: To improve the diagnosis and management ofDuchenne/Becker muscular dystrophy (DMD/BMD) in Southwest ofChina.
Methods: Clinical features of294DMD/BMD cases from Dec.2009to Dec.2011were collected. Genomic DNA was extracted using standardprocedures from the peripheral blood leukocytes, and multiple polymerasechain reaction (mPCR)+short tandem repeat (STR) were applied to detectDystrophin gene to identify genetic mutation.17cases weregastrocnemius muscle biopsies whose deletion mutations were notidentified. The standard procedure of cross-culture adaptation was used todevelop the Chinese version Pediatric Quality of Life InventoryTM3.0Neuromuscular Module (PedsQLTM3.0NMM).39patients and theirparents were investigated with the Chinese version scale. A11the patientswere followed up.
Results: Among the294cases, exons and STR deletion ofDystrophin were detected in96cases (32.65%). And exons deletions incentral hot spot of recombination (exon44-51) were detected in82cases(85.42%). Among the17cases of gastrocnemius muscle biopsies,DMD/BMD was identified in14cases (82.35%).39questionnaires of theChinese version PedsQLTM3.0NMM were completed. It had acceptablepsychometric properties. The rates of item missing were less than1%.Total and all dimension's Cronbach alpha coefficients were larger than0.7and the intraclass correlation coefficient (ICC) was larger than0.6. Theitem in the PedsQLTM3.0NMM was moderately and highly related withcorrelation coefficient of Spearman in its belonged domains than in otherdomains. Confirmatory factor analysis showed that the main indices ofgoodness of fit comparative fit index (CFI) were larger than0.9. Thescores in the cases with different disease severity had significantdifferences (P <0.05), and scores in communication and familydimensions were very low (P>0.05). Among the101DMD cases, only18were used prednisone with0.75mg/kg/d.
Conclusion: The incidence of DMD/BMD is not uncommon inSouthwest of China. Application of mPCR+STR can improve theDystrophin gene detection accuracy, but the positive rate is low. Musclebiopsy is a good supplementary means to identify the disease. The mainfactor affecting quality of life (QOL) of DMD/BMD patients is illness severity. DMD/BMD patients are generally lack of good communicationand are bad family economy. It is very urgent to develop the diagnosis andmanagement of DMD/BMD guideline suitable for China's nationalconditions.
PART II PREDNISON TREATMENT FOR DUCHENNEMUSCULAR DYSTROPHY IN SOUTHWEST OF CHINA
Objective: Duchenne muscular dystrophy (DMD) is an X-linkedrecessive disorder and is the most common form of muscular dystrophy.Glucocorticoids (GCs) are the only medication currently available thatslows the decline in muscle strength and function in patients with DMD.But it is not very clear which GCs to choose, when to initiate treatment,and how best to monitor manage side-efects. To perform a prospective,randomized, and controlled clinical trial comparing muscle strength,function, and quality of life (QoL) in patients with DMD under theguidelines in Southwest of China.
Methods:66patients with DMD (aged4-12years) from Dec.2010 to Dec.2012were divided into GC group and the controls, receivingprednisone0.75mg/kg/d for6to12months, were evaluated by MedicalResearch Council Scale (MRC), the Chinese version Pediatric Quality ofLife InventoryTM3.0Neuromuscular Module (PedsQLTM3.0NMM),high-frequency ultrasound, and creatine kinase (CK). At the same time,side-efects were monitored.
Results: For the patients in GC group after6to12months treatment,the muscle strength and function remained stable, which the differenceswere not significant (P>0.05) comparing to before treatment. Echoweakened and subcutaneous fat thickness reduced in high-frequencyultrasound, QoL improved, which the differences were significant (P <0.05) comparing to before treatment and with the controls. After6monthstreatment, the CK levels were significantly higher than those before and thecontrols, which the differences were significant (P <0.05). Thereafter CKlevels gradually declined, but they did not return to the normal ranges. Allthe results of the controls decreased significantly with the progression ofthe disease, which the differences were statistically significant (P <0.05)comparing to before treatment. Side-effects of most patients with DMDwere not significant in the clinical trial process. The weight, height, andblood pressure in the two groups were not different significantly (P>0.05).
Conclusion: Prednisone0.75mg/kg/d is suitable for the patients withDMD in Southwest of China for medium-and long-term treatment, and it is fewer side-effects. The change of CK levels is valuable for DMD diagnosis.But it is no stable and can not be used as the objective indicators to assessdrug efficacy. High-frequency ultrasound not only be able to provide anobjective basis for the early diagnosis of the disease and disease follow-up,but also be as objective indicators to assess the drug efficacy of DMD.These results will provide constructive guidance for early diagnosis, earlyintervention and effective evaluation and side-effects monitoring of DMDin the southwest of China.
PART III THE EXPRESSION AND CLINICALSIGNIFICANCE OF MUSCLE-SPECIFIC MICRORNASIN SERUM OF DUCHENNE MUSCULAR DYSTROPHY
Objective: Duchenne muscular dystrophy (DMD) is a lethalX-linked disorder caused by mutations in the dystrophin gene, whichencodes a cytoskeletal protein, dystrophin. Creatine kinase (CK) isgenerally used as a blood-based biomarker for muscular disease includingDMD, but it is not always reliable since it is easily affected by stress to the body, such as exercise. Therefore, more reliable biomarkers of musculardystrophy have long been desired. MicroRNAs (miRNAs) are small,～22nucleotide, noncoding RNAs which play important roles in the regulationof gene expression at the post-transcriptional level. Recently, it has beenreported that miRNAs exist in blood. In this study, we hypothesized thatthe expression levels of specific serum circulating miRNAs may be usefulto monitor the pathological progression of muscular diseases, andtherefore explored the possibility of these miRNAs as new biomarkers forDMD.
Methods: Using SYBR green real-time quantitative reversetranscription-PCR, we detected the expression of muscle-specific miRNAsin39patients with DMD and36normal control subjects from Dec.2010to Dec.2012. Furthermore, we analyzed the associations betweenmuscle-specific miRNAs expression and clinical features of patients withDMD comparing to creatine kinase (CK).
Results: Muscle-specific miRNAs, especial miRNA-206, weresignificantly overexpressed in serum of DMD relative to normal control (P<0.01), and its sensitivity and specificity were significantly higher than CKlevels, as a blood-based biomarker for DMD. Unlike CK levels, expressionlevels of these miRNAs in serum of DMD are little influenced by gender,age, exercise, and stress. They were highly correlated with the differentstages of DMD progression.
Conclusion: Muscle-specific miRNAs, especial miRNA-206, are newand valuable biomarkers for the diagnosis of DMD and possibly also as anew target for therapeutic interventions in humans. It provides a new ideato explore effective cure for DMD and has important implications forguidance related basic and clinical research.
目的：分析西南地区假肥大型肌营养不良（Duchenne/Beckermuscular dystrophy, DMD/BMD）患儿的诊治现状及生存质量（qualityof life, QoL）状况，以提高其诊疗水平。
方法：选取2009年12月～2011年12月我院疑诊DMD/BMD病例294例，采用多重聚合酶链反应（multiplepolymerase chain reaction, mPCR）+短串联重复序列（short tandemrepeat, STR）多态性连锁分析法行dystrophin基因检测。随机抽取17例基因检测阴性患儿行腓肠肌活检，光镜下观察病理改变。遵循量表引进程序，首次将儿童生存质量量表PedsQLTM3.0神经肌肉疾病模块（pediatric quality of life inventoryTM3.0neuromuscular module,PedsQLTM3.0NMM）翻译、修订并形成PedsQLTM3.0NMM中文版后，随机抽取39例确诊DMD/BMD患儿及其家长，调查其QoL。并随访治疗情况。
结果：294例疑诊病例mPCR+STR法检出外显子/STR缺失的阳性率为32.65%。外显子48、50和STR49CA、50CA缺失率最高。17例活检的阳性率为82.35%。PedsQLTM3.0NMM中文版的缺失率<1%，克伦巴赫系数α（Cronbach’s α）>0.7，评定者间的信度（intraclasscorrelation coefficient, ICC）>0.6，各条目与所属维度的Spearman相关系数（correlation coefficient of Spearman, rs）呈中、高度相关，相对拟合指数（Comparative fit index, CFI）>0.9。DMD/BMD患儿病情越重，疾病维度评分越低，差异有显著性（P <0.05）；而交流、家庭维度评分普遍较低，差异无统计学意义（P>0.05）。101例确诊的DMD患儿中，17.82%予口服泼尼松0.75mg/kg/d治疗，大部分未予治疗，观察随访中。
结论：mPCR+STR法可提高dystrophin基因检测的准确性，但阳性率不高；肌肉活检是确诊手段的良好补充。PedsQLTM3.0NMM中文版具有较好的信度和效度，可应用于我国DMD/BMD患儿QoL的评价。病情轻重是影响DMD/BMD患儿QoL的主要因素；他们普遍缺乏良好的交流，家庭经济条件较差。大部分DMD患儿未予治疗。制定适合我国国情的DMD/BMD临床诊治指南尤为迫切。
第二部分糖皮质激素治疗Duchenne型肌营养不良的临床疗效观察
目的：参考国外Duchenne型肌营养不良（Duchenne musculardystrophy, DMD）的临床诊治指南，多学科协作，进行糖皮质激素（glucocorticoid，GC）治疗DMD的前瞻性、随机、对照的临床试验，系统地评估GC治疗DMD的疗效和远期预后的影响，以期为西南地区DMD的早期诊断、早期干预、疗效评价及副反应监测提供建设性的指导意见。
方法：选取2010年12月～2012年12月我院经基因或病理检测确诊为DMD患儿66例，按随机数字表法随机抽取进入泼尼松组（36例）和对照组（30例）。为期6～12月泼尼松0.75mg/kg/d治疗的前瞻性临床试验后，比较治疗前后DMD患儿的肌力、肌肉功能、肌酸激酶（creatine kinase, CK）值、骨骼肌B超、生存质量（qualityof life, QoL）的变化，并监测药物副反应。
结果：治疗6～12月后，泼尼松组患儿的肌力、肌肉功能基本保持稳定并有好转，与治疗前比较，差异无显著性（P>0.05）。骨骼肌B超回声减弱，皮下脂肪厚度降低；QoL明显改善，与治疗前及同时点对照组比较，差异有显著性（P <0.05）。治疗6月后，泼尼松组患儿的CK值明显增高，与治疗前及同时点对照组比较，差异有显著性（P <0.05）；此后CK值逐渐下降，但尚未恢复正常范围。对照组患儿随病情的进展，肌力、肌肉功能下降明显，骨骼肌B超回声及QoL有明显恶化趋势，CK值有逐渐下降趋势，与治疗前比较，差异有显著性（P <0.05）。临床试验过程中，大部分DMD患儿无明显副反应，治疗前后，两组DMD患儿的体重、身高、血压（舒张压）相比较，差异均无显著性（P>0.05）。
结论：泼尼松0.75mg/kg/d适合西南地区DMD患儿的中长期治疗，副反应少。DMD患儿血清CK值的变化具有诊断价值，但存在个体差异，且受多种因素的影响，无法作为评估药物疗效的客观指标。骨骼肌B超不但能判断肌肉病变部位、范围、程度，为疾病的早期诊断、鉴别诊断及病情随访提供客观依据；而且动态监测骨骼肌B超的变化可作为评估DMD药物疗效的客观指标。这些结果可初步为西南地区DMD的早期诊断、早期干预、疗效评价及副反应监测提供建设性的指导意见。
第三部分肌肉特异性microRNAs在Duchenne型肌营养不良中的表达及意义
目的：通过检测Duchenne型肌营养不良（Duchenne musculardystrophy, DMD）血清肌肉特异性microRNAs（miRNAs）的表达水平，探讨其与DMD病情严重程度的相关性，以期作为DMD诊断的新型生物标志物。
方法：2010年12月～2012年12月我院经基因或病理检测确诊的DMD患儿66例，按随机数字表法随机抽取进入实验组（39例）。于同一时期本院儿童保健科健康体检的健康男性儿童中抽取对照组36例。行肌力、肌肉功能、生存质量（quality of life, QoL）评估及血清肌酸激酶（creatine kinase, CK）检测后，采用实时定量荧光聚合酶链反应（Real-time quantitative polymerase chain reaction,RT-qPCR）法检测血清肌肉特异性miRNAs的表达水平，并与血清CK值及肌力、肌肉功能和QoL进行比较，探讨肌肉特异性miRNAs与DMD病情严重程度的相关性。
结果：实验组血清肌肉特异性miRNAs的表达水平较对照组明显增高，差异有显著性（P <0.01）。肌肉特异性miRNAs与年龄之间无相关性（P>0.05），与肌力、肌肉功能及QoL之间存在显著的负相关性（P <0.05），尤以miRNA-206为显著。CK值与年龄之间存在显著的相关性（P <0.05），与肌力、肌肉功能及QoL无相关性（P>0.05）。受试者工作特征（receiver operating characteristic, ROC）曲线分析显示，肌肉特异性miRNAs的AUC≥0.9，诊断价值较高，灵敏度和特异度好，尤以miRNA-206为著。
结论：肌肉特异性miRNAs在DMD患儿的血清中高度表达，稳定性强，并与病情的严重程度密切相关，可作为DMD诊断的新型生物标志物，对指导相关基础与临床研究具有重要的意义。
PART I DUCHENNE/BECKER MUSCULARDYSTROPHY IN SOUTHWEST OF CHINA
Objective: To improve the diagnosis and management ofDuchenne/Becker muscular dystrophy (DMD/BMD) in Southwest ofChina.
Methods: Clinical features of294DMD/BMD cases from Dec.2009to Dec.2011were collected. Genomic DNA was extracted using standardprocedures from the peripheral blood leukocytes, and multiple polymerasechain reaction (mPCR)+short tandem repeat (STR) were applied to detectDystrophin gene to identify genetic mutation.17cases weregastrocnemius muscle biopsies whose deletion mutations were notidentified. The standard procedure of cross-culture adaptation was used todevelop the Chinese version Pediatric Quality of Life InventoryTM3.0Neuromuscular Module (PedsQLTM3.0NMM).39patients and theirparents were investigated with the Chinese version scale. A11the patientswere followed up.
Results: Among the294cases, exons and STR deletion ofDystrophin were detected in96cases (32.65%). And exons deletions incentral hot spot of recombination (exon44-51) were detected in82cases(85.42%). Among the17cases of gastrocnemius muscle biopsies,DMD/BMD was identified in14cases (82.35%).39questionnaires of theChinese version PedsQLTM3.0NMM were completed. It had acceptablepsychometric properties. The rates of item missing were less than1%.Total and all dimension's Cronbach alpha coefficients were larger than0.7and the intraclass correlation coefficient (ICC) was larger than0.6. Theitem in the PedsQLTM3.0NMM was moderately and highly related withcorrelation coefficient of Spearman in its belonged domains than in otherdomains. Confirmatory factor analysis showed that the main indices ofgoodness of fit comparative fit index (CFI) were larger than0.9. Thescores in the cases with different disease severity had significantdifferences (P <0.05), and scores in communication and familydimensions were very low (P>0.05). Among the101DMD cases, only18were used prednisone with0.75mg/kg/d.
Conclusion: The incidence of DMD/BMD is not uncommon inSouthwest of China. Application of mPCR+STR can improve theDystrophin gene detection accuracy, but the positive rate is low. Musclebiopsy is a good supplementary means to identify the disease. The mainfactor affecting quality of life (QOL) of DMD/BMD patients is illness severity. DMD/BMD patients are generally lack of good communicationand are bad family economy. It is very urgent to develop the diagnosis andmanagement of DMD/BMD guideline suitable for China's nationalconditions.
PART II PREDNISON TREATMENT FOR DUCHENNEMUSCULAR DYSTROPHY IN SOUTHWEST OF CHINA
Objective: Duchenne muscular dystrophy (DMD) is an X-linkedrecessive disorder and is the most common form of muscular dystrophy.Glucocorticoids (GCs) are the only medication currently available thatslows the decline in muscle strength and function in patients with DMD.But it is not very clear which GCs to choose, when to initiate treatment,and how best to monitor manage side-efects. To perform a prospective,randomized, and controlled clinical trial comparing muscle strength,function, and quality of life (QoL) in patients with DMD under theguidelines in Southwest of China.
Methods:66patients with DMD (aged4-12years) from Dec.2010 to Dec.2012were divided into GC group and the controls, receivingprednisone0.75mg/kg/d for6to12months, were evaluated by MedicalResearch Council Scale (MRC), the Chinese version Pediatric Quality ofLife InventoryTM3.0Neuromuscular Module (PedsQLTM3.0NMM),high-frequency ultrasound, and creatine kinase (CK). At the same time,side-efects were monitored.
Results: For the patients in GC group after6to12months treatment,the muscle strength and function remained stable, which the differenceswere not significant (P>0.05) comparing to before treatment. Echoweakened and subcutaneous fat thickness reduced in high-frequencyultrasound, QoL improved, which the differences were significant (P <0.05) comparing to before treatment and with the controls. After6monthstreatment, the CK levels were significantly higher than those before and thecontrols, which the differences were significant (P <0.05). Thereafter CKlevels gradually declined, but they did not return to the normal ranges. Allthe results of the controls decreased significantly with the progression ofthe disease, which the differences were statistically significant (P <0.05)comparing to before treatment. Side-effects of most patients with DMDwere not significant in the clinical trial process. The weight, height, andblood pressure in the two groups were not different significantly (P>0.05).
Conclusion: Prednisone0.75mg/kg/d is suitable for the patients withDMD in Southwest of China for medium-and long-term treatment, and it is fewer side-effects. The change of CK levels is valuable for DMD diagnosis.But it is no stable and can not be used as the objective indicators to assessdrug efficacy. High-frequency ultrasound not only be able to provide anobjective basis for the early diagnosis of the disease and disease follow-up,but also be as objective indicators to assess the drug efficacy of DMD.These results will provide constructive guidance for early diagnosis, earlyintervention and effective evaluation and side-effects monitoring of DMDin the southwest of China.
PART III THE EXPRESSION AND CLINICALSIGNIFICANCE OF MUSCLE-SPECIFIC MICRORNASIN SERUM OF DUCHENNE MUSCULAR DYSTROPHY
Objective: Duchenne muscular dystrophy (DMD) is a lethalX-linked disorder caused by mutations in the dystrophin gene, whichencodes a cytoskeletal protein, dystrophin. Creatine kinase (CK) isgenerally used as a blood-based biomarker for muscular disease includingDMD, but it is not always reliable since it is easily affected by stress to the body, such as exercise. Therefore, more reliable biomarkers of musculardystrophy have long been desired. MicroRNAs (miRNAs) are small,～22nucleotide, noncoding RNAs which play important roles in the regulationof gene expression at the post-transcriptional level. Recently, it has beenreported that miRNAs exist in blood. In this study, we hypothesized thatthe expression levels of specific serum circulating miRNAs may be usefulto monitor the pathological progression of muscular diseases, andtherefore explored the possibility of these miRNAs as new biomarkers forDMD.
Methods: Using SYBR green real-time quantitative reversetranscription-PCR, we detected the expression of muscle-specific miRNAsin39patients with DMD and36normal control subjects from Dec.2010to Dec.2012. Furthermore, we analyzed the associations betweenmuscle-specific miRNAs expression and clinical features of patients withDMD comparing to creatine kinase (CK).
Results: Muscle-specific miRNAs, especial miRNA-206, weresignificantly overexpressed in serum of DMD relative to normal control (P<0.01), and its sensitivity and specificity were significantly higher than CKlevels, as a blood-based biomarker for DMD. Unlike CK levels, expressionlevels of these miRNAs in serum of DMD are little influenced by gender,age, exercise, and stress. They were highly correlated with the differentstages of DMD progression.
Conclusion: Muscle-specific miRNAs, especial miRNA-206, are newand valuable biomarkers for the diagnosis of DMD and possibly also as anew target for therapeutic interventions in humans. It provides a new ideato explore effective cure for DMD and has important implications forguidance related basic and clinical research.
引文
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