An Engraftable Human Embryonic Stem Cell Neuronal Lineage-Specific Derivative Retains Embryonic Chromatin Plasticity for Scale-Up CNS Regeneration.
An Engraftable Human Embryonic Stem Cell Neuronal Lineage-Specific Derivative Retains Embryonic Chromatin Plasticity for Scale-Up CNS Regeneration.
复制标题
DOI：
10.7243/2050-1218-1-3
复制
发表时间：
2012-09
期刊：
影响因子：
--
通讯作者：
Xuejun H. Parsons
中科院分区：
文献类型：
--
作者：
Xuejun H. Parsons
背景多能人胚胎干细胞（hESC）通过在发育中的CNS中提供用于修复的人神经元细胞类型的多样性来提供对广泛的神经障碍的治疗。然而，实现hESC衍生物的治疗潜力受到了通过不可控和低效的多谱系分化从多能细胞产生神经元细胞的阻碍。以前，我们使用了一个确定的平台来鉴定视黄酸，视黄酸足以直接从hESC的多能状态诱导神经外胚层的特化，并在发育中的CNS中高效地触发均匀的神经元谱系特异性进展为人神经元祖细胞（hESC-I hNuP）和神经元（hESC-I hNus）。方法在实现了多能hESC向神经元谱系的均匀转化后，在本研究中，检测了hESC-1 hNuPs中一组染色质修饰剂的表达和细胞内分布模式，并与两种原型神经上皮样人神经干细胞（hNSCs）进行了比较，所述两种原型神经上皮样人神经干细胞（hNSCs）来源于hESC或直接从人胎儿神经外胚层体内分离。这些hESC-1 hNuPs表达高水平的活性染色质修饰剂，包括乙酰化组蛋白H3和H4，HDAC 1，Brg-1和hSNF 2 H，保留胚胎乙酰化的全局活性染色质状态。与这一观察结果一致的是，几种调节组蛋白H3 K9甲基化的抑制性染色质重塑因子，包括SIRT 1、SUV 39 H1和Brm，在hESC-1 hNuPs中是无活性的。这些Nurr 1阳性hESC-I hNuP不表达典型的hNSC标记物，有效地且排他地产生神经元，因为它们不分化成神经胶质细胞。在脑中植入后，hESC-I hNuP以高流行率产生良好分散和良好整合的人神经元。结论这些观察结果表明，与原型神经上皮样巢蛋白阳性hNSC不同，这些体外神经外胚层来源的Nurr 1阳性hESC-I hNuPs是一种更具神经元谱系特异性和可塑性的人类干细胞衍生物，提供高纯度的可移植的人胚胎神经元祖细胞，并大量供应，其具有足够的神经原性潜力，将中枢神经系统再生作为干细胞疗法转化为临床试验中的患者。
BACKGROUND Pluripotent human embryonic stem cells (hESCs) proffer cures for a wide range of neurological disorders by supplying the diversity of human neuronal cell types in the developing CNS for repair. However, realizing the therapeutic potential of hESC derivatives has been hindered by generating neuronal cells from pluripotent cells through uncontrollable and inefficient multi-lineage differentiation. Previously, we used a defined platform to identify retinoic acid as sufficient to induce the specification of neuroectoderm direct from the pluripotent state of hESCs and trigger uniform neuronal lineage-specific progression to human neuronal progenitors (hESC-I hNuPs) and neurons (hESC-I hNus) in the developing CNS with high efficiency. METHODS Having achieved uniformly conversion of pluripotent hESCs to a neuronal lineage, in this study, the expression and intracellular distribution patterns of a set of chromatin modifiers in hESC-I hNuPs were examined and compared to the two prototypical neuroepithelial-like human neural stem cells (hNSCs) either derived from hESCs or isolated directly from the human fetal neuroectoderm in vivo. RESULTS These hESC-I hNuPs expressed high levels of active chromatin modifiers, including acetylated histone H3 and H4, HDAC1, Brg-1, and hSNF2H, retaining an embryonic acetylated globally active chromatin state. Consistent with this observation, several repressive chromatin remodeling factors regulating histone H3K9 methylation, including SIRT1, SUV39H1, and Brm, were inactive in hESC-I hNuPs. These Nurr1-positive hESC-I hNuPs, which did not express the canonical hNSC markers, yielded neurons efficiently and exclusively, as they did not differentiate into glial cells. Following engraftment in the brain, hESC-I hNuPs yielded well-dispersed and well-integrated human neurons at a high prevalence. CONCLUSIONS These observations suggest that, unlike the prototypical neuroepithelial-like nestin-positive hNSCs, these in vitro neuroectoderm-derived Nurr1-positive hESC-I hNuPs are a more neuronal lineage-specific and plastic human stem cell derivative, providing an engraftable human embryonic neuronal progenitor in high purity and large supply with adequate neurogenic potential for scale-up CNS regeneration as stem cell therapy to be translated to patients in clinical trials.