Directed differentiation of patient-specific induced pluripotent stem cells identifies the transcriptional repression and epigenetic modification of NKX2-5, HAND1, and NOTCH1 in hypoplastic left heart syndrome.
Directed differentiation of patient-specific induced pluripotent stem cells identifies the transcriptional repression and epigenetic modification of NKX2-5, HAND1, and NOTCH1 in hypoplastic left heart syndrome.
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DOI：
10.1371/journal.pone.0102796
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发表时间：
2014
期刊：
影响因子：
3.7
通讯作者：
Oh H
中科院分区：
文献类型：
--
作者：
Kobayashi J;Yoshida M;Tarui S;Hirata M;Nagai Y;Kasahara S;Naruse K;Ito H;Sano S;Oh H
左心发育不全综合征(HLHS)的遗传学基础尚不清楚，缺乏重建心脏发育不良的动物模型阻碍了对该疾病的研究。这项研究通过使用疾病特异性诱导多能干细胞(IPS)，研究了转录和表观遗传程序的改变控制，这可能影响HLHS的发展。从先天性心脏病患者中分离出心脏祖细胞(CPC)，以产生患者特有的iPS细胞。对HLHS和BV心脏来源的iPS细胞进行了基因表达的比较分析，以剖析可能促进疾病表型的复杂遗传电路。HLHS和BV心脏来源的CPC都被重新编程，以产生疾病特异性的iPS细胞，这些细胞显示出典型的人类胚胎干细胞特征，表达多潜能标记，并可以分化为心肌细胞。然而，HLHS-iPS细胞的心肌分化潜能低于BV-iPS细胞。定量基因表达分析显示，与对照细胞相比，HLHS来源的iPS细胞在转录水平上抑制了NKX2-5的表达，降低了Tbx2和Noch/嘿信号的水平，并抑制了HAND1/2的转录。尽管HLHS来源的CPC和iPS细胞与BV来源的细胞相比，SRE和TNNT2的转录活性降低，但NKX2-5、HAND1和NOTCH1共同转染HLHS来源的细胞后，这些启动子的激活得到协同恢复。值得注意的是，功能获得和功能丧失研究表明，NKX2-5对NPPA转录激活有显著影响。此外，分化的HLHS来源的iPS细胞显示H3K4二甲基化和组蛋白H3乙酰化减少，但H3K27三甲基化增加，以抑制NKX2-5启动子上的转录激活。这些发现表明，患者特有的iPS细胞可能至少部分通过NKX2-5、HAND1和NOTCH1的组合表达，为复杂的转录和表观遗传机制提供分子洞察力，这些基因协同导致HLHS心脏畸形。
The genetic basis of hypoplastic left heart syndrome (HLHS) remains unknown, and the lack of animal models to reconstitute the cardiac maldevelopment has hampered the study of this disease. This study investigated the altered control of transcriptional and epigenetic programs that may affect the development of HLHS by using disease-specific induced pluripotent stem (iPS) cells. Cardiac progenitor cells (CPCs) were isolated from patients with congenital heart diseases to generate patient-specific iPS cells. Comparative gene expression analysis of HLHS- and biventricle (BV) heart-derived iPS cells was performed to dissect the complex genetic circuits that may promote the disease phenotype. Both HLHS- and BV heart-derived CPCs were reprogrammed to generate disease-specific iPS cells, which showed characteristic human embryonic stem cell signatures, expressed pluripotency markers, and could give rise to cardiomyocytes. However, HLHS-iPS cells exhibited lower cardiomyogenic differentiation potential than BV-iPS cells. Quantitative gene expression analysis demonstrated that HLHS-derived iPS cells showed transcriptional repression of NKX2-5, reduced levels of TBX2 and NOTCH/HEY signaling, and inhibited HAND1/2 transcripts compared with control cells. Although both HLHS-derived CPCs and iPS cells showed reduced SRE and TNNT2 transcriptional activation compared with BV-derived cells, co-transfection of NKX2-5, HAND1, and NOTCH1 into HLHS-derived cells resulted in synergistic restoration of these promoters activation. Notably, gain- and loss-of-function studies revealed that NKX2-5 had a predominant impact on NPPA transcriptional activation. Moreover, differentiated HLHS-derived iPS cells showed reduced H3K4 dimethylation as well as histone H3 acetylation but increased H3K27 trimethylation to inhibit transcriptional activation on the NKX2-5 promoter. These findings suggest that patient-specific iPS cells may provide molecular insights into complex transcriptional and epigenetic mechanisms, at least in part, through combinatorial expression of NKX2-5, HAND1, and NOTCH1 that coordinately contribute to cardiac malformations in HLHS.
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DOI：
10.1016/s0022-5223(02)73575-7
发表时间：
2003-08-01
影响因子：
6
作者：
Sano, S;Ishino, K;Ohtsuki, S
通讯作者：
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影响因子：
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14.8
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影响因子：
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作者：
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Giordano, Emanuele