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c-MYC 非依赖性核重编程有利于诱导多能干细胞的心脏生成潜能。

c-MYC independent nuclear reprogramming favors cardiogenic potential of induced pluripotent stem cells.

机构信息

Division of Cardiovascular Diseases, Departments of Medicine, Mayo Clinic, Rochester, MN, USA.

出版信息

J Cardiovasc Transl Res. 2010 Feb;3(1):13-23. doi: 10.1007/s12265-009-9150-5.


DOI:10.1007/s12265-009-9150-5
PMID:20221419
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC2835311/
Abstract

Induced pluripotent stem cell (iPS) technology has launched a new platform in regenerative medicine aimed at deriving unlimited replacement tissue from autologous sources through somatic cell reprogramming using stemness factor sets. In this way, authentic cardiomyocytes have been obtained from iPS and recently demonstrated in proof-of-principle studies to repair infarcted heart. Optimizing the cardiogenic potential of iPS progeny would ensure a maximized yield of bioengineered cardiac tissue. Here, we reprogrammed fibroblasts in the presence or absence of c-MYC to determine if the acquired cardiogenicity is sensitive to the method of nuclear reprogramming. Using lentiviral constructs that expressed stemness factors SOX2, OCT4, and KLF4 with or without c-MYC, iPS clones generated through fibroblast reprogramming demonstrated indistinguishable characteristics for 5 days of differentiation with similar cell morphology, growth rates, and chimeric embryo integration. However, 4-factor c-MYC dependent nuclear reprogramming produced iPS progeny that consistently prolonged the expression of pluripotent Oct-4 and Fgf4 genes and repressed cardiac differentiation. In contrast, 3-factor c-MYC-less iPS clones efficiently up-regulated pre-cardiac (CXCR4, Flk-1, and Mesp1/2) and cardiac (Nkx2.5, Mef2c, and Myocardin) gene expression patterns. In fact, 3-factor iPS progeny demonstrated early and robust cardiogenesis during in vitro differentiation with consistent beating activity, sarcomere maturation, and rhythmical intracellular calcium dynamics. Thus, nuclear reprogramming independent of c-MYC enhances production of pluripotent stem cells with innate cardiogenic potential.

摘要

诱导多能干细胞(iPS)技术在再生医学领域开创了一个新的平台,旨在通过使用干性因子集对自体来源的体细胞进行重编程,从而获得无限量的替代组织。通过这种方式,已经从 iPS 中获得了真正的心肌细胞,并在最近的原理验证研究中证明可修复梗死的心脏。优化 iPS 后代的成心肌潜能将确保生物工程心脏组织的产量最大化。在这里,我们在存在或不存在 c-MYC 的情况下重编程成纤维细胞,以确定获得的成心肌性是否对核重编程方法敏感。使用表达干性因子 SOX2、OCT4 和 KLF4 的慢病毒构建体,带有或不带有 c-MYC,通过成纤维细胞重编程产生的 iPS 克隆在 5 天的分化过程中表现出相似的细胞形态、生长速度和嵌合胚胎整合,具有不可区分的特征。然而,4 因子 c-MYC 依赖性核重编程产生的 iPS 后代持续延长多能 Oct-4 和 Fgf4 基因的表达,并抑制心脏分化。相比之下,3 因子 c-MYC 缺失的 iPS 克隆有效地上调了前心肌(CXCR4、Flk-1 和 Mesp1/2)和心肌(Nkx2.5、Mef2c 和 Myocardin)基因表达模式。事实上,3 因子 iPS 后代在体外分化过程中表现出早期和强大的成心肌性,具有一致的搏动活性、肌节成熟和节律性细胞内钙动力学。因此,不依赖 c-MYC 的核重编程增强了具有固有成心肌潜能的多能干细胞的产生。

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本文引用的文献

[1]
Induced pluripotent reprogramming from promiscuous human stemness related factors.

Clin Transl Sci. 2009-4

[2]
Stem cell platforms for regenerative medicine.

Clin Transl Sci. 2009-6

[3]
Enhanced generation of induced pluripotent stem cells from a subpopulation of human fibroblasts.

PLoS One. 2009-9-23

[4]
Strategies for therapeutic repair: The "R(3)" regenerative medicine paradigm.

Clin Transl Sci. 2008-9

[5]
Generation of pluripotent stem cells from patients with type 1 diabetes.

Proc Natl Acad Sci U S A. 2009-9-15

[6]
iPS programmed without c-MYC yield proficient cardiogenesis for functional heart chimerism.

Circ Res. 2009-9-25

[7]
Modelling pathogenesis and treatment of familial dysautonomia using patient-specific iPSCs.

Nature. 2009-9-17

[8]
Repair of acute myocardial infarction by human stemness factors induced pluripotent stem cells.

Circulation. 2009-8-4

[9]
Elite and stochastic models for induced pluripotent stem cell generation.

Nature. 2009-7-2

[10]
Disease-corrected haematopoietic progenitors from Fanconi anaemia induced pluripotent stem cells.

Nature. 2009-7-2

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