Impact of ANKRD1 mutations associated with hypertrophic cardiomyopathy on contraction parameters of engineered heart tissue
Impact of ANKRD1 mutations associated with hypertrophic cardiomyopathy on contraction parameters of engineered heart tissue
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DOI：
10.1007/s00395-013-0349-x
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发表时间：
2013-05-01
影响因子：
9.5
通讯作者：
Carrier, Lucie
中科院分区：
文献类型：
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作者：
Crocini, Claudia;Arimura, Takuro;Carrier, Lucie
肥厚型心肌病（HCM）是一种与肌节基因突变相关的心肌疾病。在ANKRD1中发现了三个突变，编码锚蛋白重复结构域1（ANKRD1），一种位于肌节中的转录辅因子。在本研究中，我们研究了HCM相关ANKRD1突变的表达是否会影响基因转移后工程心脏组织（EHTs）的收缩参数。EHT由新生大鼠心脏细胞产生，并用编码GFP或myc标记的野生型（WT）或突变体（P52A、T123M或I280V）ANKRD1的腺相关病毒转导。从培养的第8天至第16天分析收缩参数，并在不存在或存在蛋白酶体抑制剂环氧霉素的情况下评价24小时。在标准条件下，只有WT-和T123M-ANKRD1正确地掺入肌节中。T123M-ANKRD1转导的EHT表现出比WT更高的收缩和舒张力和速度。P52A和I280V-ANKRD1是高度不稳定的，不掺入肌节，并且不诱导收缩改变。在环氧霉素处理后，P52A和I280V都被稳定并掺入肌节中。I280V转导的EHTs表现出延长的松弛。这些数据表明AN-KRD1突变对心肌细胞功能的不同影响：T123 M突变在所有条件下的增益功能和I280 V突变的显性负效应，其可能仅在蛋白酶体受损时起作用。
Hypertrophic cardiomyopathy (HCM) is a myocardial disease associated with mutations in sarcomeric genes. Three mutations were found in ANKRD1, encoding ankyrin repeat domain 1 (ANKRD1), a transcriptional co-factor located in the sarcomere. In the present study, we investigated whether expression of HCM-associated ANKRD1 mutations affects contraction parameters after gene transfer in engineered heart tissues (EHTs). EHTs were generated from neonatal rat heart cells and were transduced with adeno-associated virus encoding GFP or myc-tagged wild-type (WT) or mutant (P52A, T123M, or I280V) ANKRD1. Contraction parameters were analyzed from day 8 to day 16 of culture, and evaluated in the absence or presence of the proteasome inhibitor epoxomicin for 24 h. Under standard conditions, only WT- and T123M-ANKRD1 were correctly incorporated in the sarcomere. T123M-ANKRD1-transduced EHTs exhibited higher force and velocities of contraction and relaxation than WT- P52A- and I280V-ANKRD1 were highly unstable, not incorporated into the sarcomere, and did not induce contractile alterations. After epoxomicin treatment, P52A and I280V were both stabilized and incorporated into the sarcomere. I280V-transduced EHTs showed prolonged relaxation. These data suggest different impacts of AN-KRD1 mutations on cardiomyocyte function: gain-offunction for T123M mutation under all conditions and dominant-negative effect for the I280V mutation which may come into play only when the proteasome is impaired.