Multicellular Transcriptional Analysis of Mammalian Heart Regeneration.
Multicellular Transcriptional Analysis of Mammalian Heart Regeneration.
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DOI：
10.1161/circulationaha.117.028252
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发表时间：
2017-09-19
期刊：
影响因子：
37.8
通讯作者：
Porrello ER
中科院分区：
文献类型：
--
作者：
Quaife-Ryan GA;Sim CB;Ziemann M;Kaspi A;Rafehi H;Ramialison M;El-Osta A;Hudson JE;Porrello ER
补充数字内容可在文本中找到。成年哺乳动物心脏损伤后不能再生是心血管医学的一个主要障碍。相比之下，新生哺乳动物的心脏保留了短暂的再生能力，在出生后不久就失去了。确定新生儿期再生能力的分子机制仍然是心脏生物学的中心目标。在这里，我们在出生后发育和损伤后组装了多个心脏细胞群的转录组框架，这使得第一次能够比较分析再生（新生儿）与非再生（成人）状态。心肌细胞，成纤维细胞，白细胞和内皮细胞从梗死和非梗死新生儿（P1）和成年（P56）小鼠心脏分离酶解离和荧光激活细胞分选术后第3天。对这些细胞群进行RNA测序，以生成心脏发育、修复和再生过程中主要心脏细胞群的转录组。为了补充我们的转录组数据，我们还调查了出生后成熟期间心肌细胞的表观遗传景观，通过使用纯化小鼠心肌细胞核（P1，P14和P56）的转座酶可降解染色质测定法对可接近的染色质区域进行深度测序。分析心肌细胞和非心肌细胞转录程序发现了几个损伤响应基因在再生和非再生时间点。然而，所有心脏细胞类型中的大多数转录变化是由新生儿阶段到成年期的发育成熟引起的，而不是由不同的再生特异性基因程序激活引起的。此外，成人白细胞和成纤维细胞的特点是表达的增殖基因表达网络梗死后，这反映了新生儿的状态。相比之下，心肌细胞未能重新激活新生儿增殖网络梗死后，这与出生后成熟过程中细胞周期基因周围的染色质可及性丧失有关。这项工作提供了一个全面的框架和多个心脏细胞群体在心脏发育，修复和再生的转录资源。我们的研究结果定义了一个支持新生儿再生状态的调控程序，并确定了可能限制成年心肌细胞再生程序再诱导的染色质景观的改变。
Supplemental Digital Content is available in the text. The inability of the adult mammalian heart to regenerate following injury represents a major barrier in cardiovascular medicine. In contrast, the neonatal mammalian heart retains a transient capacity for regeneration, which is lost shortly after birth. Defining the molecular mechanisms that govern regenerative capacity in the neonatal period remains a central goal in cardiac biology. Here, we assemble a transcriptomic framework of multiple cardiac cell populations during postnatal development and following injury, which enables comparative analyses of the regenerative (neonatal) versus nonregenerative (adult) state for the first time. Cardiomyocytes, fibroblasts, leukocytes, and endothelial cells from infarcted and noninfarcted neonatal (P1) and adult (P56) mouse hearts were isolated by enzymatic dissociation and fluorescence-activated cell sorting at day 3 following surgery. RNA sequencing was performed on these cell populations to generate the transcriptome of the major cardiac cell populations during cardiac development, repair, and regeneration. To complement our transcriptomic data, we also surveyed the epigenetic landscape of cardiomyocytes during postnatal maturation by performing deep sequencing of accessible chromatin regions by using the Assay for Transposase-Accessible Chromatin from purified mouse cardiomyocyte nuclei (P1, P14, and P56). Profiling of cardiomyocyte and nonmyocyte transcriptional programs uncovered several injury-responsive genes across regenerative and nonregenerative time points. However, the majority of transcriptional changes in all cardiac cell types resulted from developmental maturation from neonatal stages to adulthood rather than activation of a distinct regeneration-specific gene program. Furthermore, adult leukocytes and fibroblasts were characterized by the expression of a proliferative gene expression network following infarction, which mirrored the neonatal state. In contrast, cardiomyocytes failed to reactivate the neonatal proliferative network following infarction, which was associated with loss of chromatin accessibility around cell cycle genes during postnatal maturation. This work provides a comprehensive framework and transcriptional resource of multiple cardiac cell populations during cardiac development, repair, and regeneration. Our findings define a regulatory program underpinning the neonatal regenerative state and identify alterations in the chromatin landscape that could limit reinduction of the regenerative program in adult cardiomyocytes.