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Distinguishing Between Mouse and Human Pluripotent Stem Cell Regulation: the Best Laid Plans of Mice and Men

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Distinguishing Between Mouse and Human Pluripotent Stem Cell Regulation: the Best Laid Plans of Mice and Men EMBRYONIC STEM CELLS/INDUCED PLURIPOTENT STEM CELLS Distinguishing Between Mouse and Human Pluripotent Stem Cell Regulation: The Best Laid Plans of Mice and Men a,b a a,b ANGELIQUE SCHNERCH, CHANTAL CERDAN, MICKIE BHATIA aStem Cell and Cancer Research Institute, Faculty of Health Sciences, McMaster University, Hamilton, Ontario, Canada; bDepartment of Biochemistry and Biomedical Sciences, Faculty of Health Sciences, McMaster University, Hamilton, Ontario, Canada Key Words. Embryonic stem cells • Pluripotent stem cells • Cell biology • Cell signaling ABSTRACT Pluripotent stem cells (PSCs) have been derived from the tinguish between universally applicable and species-specific embryos of mice and humans, representing the two major features, we collated and compared the molecular and cel- sources of PSCs. These cells are universally defined by lular descriptions of mouse and human PSCs. Here we their developmental properties, specifically their self- compare and contrast the response to signals dictated by renewal capacity and differentiation potential which are the transcriptome and epigenome of mouse and human regulated in mice and humans by complex transcriptional PSCs that will hopefully act as a critical resource to the networks orchestrated by conserved transcription factors. field. These analyses underscore the importance of However, significant differences exist in the transcriptional accounting for species differences when designing strat- networks and signaling pathways that control mouse and egies to capitalize on the clinical potential of human PSCs. human PSC self-renewal and lineage development. To dis- STEM CELLS 2010;28:419–430 Disclosure of potential conflicts of interest is found at the end of this article. and stem cell maintenance was initially derived from the mu- INTRODUCTION rine system. The discovery of a conserved pluripotency net- work established by the transcription factors Oct4, Sox2, and Both hope and hype surround the potential to generate abundant Nanog lends credence to the continued use of the mouse model sources of differentiated cells for cell replacement therapies from for understanding human PSCs. However, fundamental differ- human pluripotent stem cells (PSCs), and although progress has ences in the global molecular signatures [22, 23] and signaling been made in the last decade, there is still more work to be done pathways [24] that maintain mouse and human PSCs exist. In before achieving this ultimate goal [1–3]. The clinical promise of addition, differences in colony shape, growth rate, surface PSCs versus other sources of somatic stem cells is based on two markers, and developmental potential between mESC and defining characteristics: 1) robust self-renewal capacity in vitro hESC cultures further demonstrate that distinct cellular and [4], and 2) multilineage differentiation to derivatives of the three molecular mechanisms define mouse versus human PSCs. embryonic germ layers and subsequent lineages [5]. PSCs repre- Currently, hundreds of reports have described the molecular sent a unique class of developmentally plastic cells that have basis of self-renewal and differentiation in mouse and human, yet been derived from several sources in mice and humans exempli- a thorough collation and comparison of this information is lack- fied by: fetal gonadal ridges and mesenteries (primordial germ ing. Here we provide a collective resource comparing the core cells) [6, 7], preimplantation blastocysts (embryonic stem cells; transcriptional networks and the emerging roles of microRNAs [ESCs]) [8–11], postimplantation mouse embryos (epiblast stem (miRNAs) and epigenetics in orchestrating signals that govern cells; EpiSCs) [12, 13], and, recently, induced pluripotent stem self-renewal and differentiation, and clearly reveal convergent cells (iPS cells) [14–21]. Of these, human and mouse ESCs repre- and divergent pathways that maintain mouse and human PSCs. sent the most prevalently used and studied PSCs to date, and serve as the comparative benchmark for other sources of PSCs, including reprogrammed somatic cells. Dissection of the molecular basis of multilineage differen- CONTRASTING SIGNALING PATHWAYS IN tiation and self-renewal in mouse and human PSCs may pro- MOUSE AND HUMAN PSCS vide insight into the current paucity of clinically useful cells arising from all sources of human PSCs. Much of our current Our understanding of the transcriptional networks associated understanding of molecular networks involved in pluripotency with self-renewal and pluripotency of mouse and human Author contributions: A.S.: Manuscript writing, collection and assembly of data, data analysis and interpretation; C.C.: Manuscript writing, collection and assembly of data; M.B.: Manuscript writing, conception and design, financial support, final approval of manuscript. A.S. and C.C. contributed equally to this work. Correspondence: Dr. Mickie Bhatia, PhD., Stem Cell and Cancer Research Institute (SCC-RI), Faculty of Health Sciences, McMaster University, 1200 Main Street West, MDCL 5029, Hamilton, Ontario, L8N 3Z5, Canada. Telephone: 905-525-9140 Extension: 28687; Fax: 905-522-7772; e-mail: [email protected] Received September 28, 2009; accepted for publication December 3, 2009; first published online in STEM CELLS EXPRESS January 6, 2010. VC AlphaMed Press 1066-5099/2009/$30.00/0 doi: 10.1002/stem.298 STEM CELLS 2010;28:419–430 www.StemCells.com 420 Mouse and Human Pluripotent Stem Cell Regulation Figure 1. Human and mouse embryonic stem cell (ESC) identity is sustained by mainly distinct signaling networks. Both FGF and IGF path- ways are central mediators in the maintenance of undifferentiated hESCs, likely through MEK/ERK [117–119] and PI3K/Akt activation. FGF2 has been reported to induce the expression of hESC maintenance factors such as transforming growth factor beta (summarized in the review arti- cle by Stewart et al. [120]). SMAD2/3 indirectly regulates OCT-4 hESCs via Activin A signaling which is mediated by SMAD2/3 [121] (indirect regulation shown by dashed arrows). In contrast, LIF/Stat3 is required for maintaining the undifferentiated state in mESCs [122]. As long as the balance remains in favor of Stat3, self-renewal is promoted at the expense of differentiation (MEK/ERK signaling pathway) [123, 124]. BMP4 can inhibit the MEK/ERK differentiation pathway resulting in mESC self-renewal [125]. Under specific chemical inhibition GSK3, FGF, and ERK signaling STAT3 is not required for mouse ES self-renewal [116] providing a groundstate for self-renewal, however, for the purposes of this review, we chose to display the canonical signaling pathways of self-renewal in ES cells. Abbreviations: ERK, extra-cellular-signal-related kinase; FGF, fibroblast growth factor; IGF, insulin-like growth factor; MEK, MAPK/ERK kinase. ESCs has increased significantly in recent years; however, in the signaling pathways controlling the stem cell state and very little is known about the crosstalk between pathways. fate exist between mouse and human PSCs, thus devising This is largely obscured by the complexity of ESC cultures, improved differentiation strategies, for instance, small mole- which may be, in part, owing to their heterogeneity [25, 26]. cule delivery [35], should be informed by and validated in The intersections between the core signaling pathways in human PSC lines. mouse and human ESCs, with emphasis on species differen- ces, have been compiled in Figure 1. In addition, the expres- sion of key components of these signaling pathways and their functions has been summarized in Table 1. Questions as to ACOMMON TRANSCRIPTIONAL HUB OF whether the dissimilarity between mouse and human ESC sig- PLURIPOTENCY? naling pathways are the result of genuine species-specific, de- velopmental stage-specific [27], or epigenetic variations have The consistent requirement of the three transcription factors: arisen. It has been reported that mouse EpiSCs, derived from Oct4, Sox2, and Nanog, for the maintenance of both mouse the postimplantation epiblast, exhibit characteristics similar to and human ESCs form the foundation of mammalian pluripo- hESCs at multiple levels (culture requirements, expression tency. In spite of the conservation of this ‘‘transcriptional profiles, transcriptional networks, and epigenetic status) [12, hub,’’ the gene targets and functional effects following modu- 13]. Interestingly, an 7-fold greater overlap in Oct4 targets lation of these factors appear to be species-specific. The func- exists between hESCs and EpiSCs compared with mESCs tional effects of gene expression changes and a summary of [13]. This suggests that EpiSCs and hESCs represent a similar the gene targets of Oct4/Sox2/Nanog in hESCs and Oct4 in developmental stage; however, progress toward modeling mESCs are discussed below in detail and presented in sup- human differentiation in EpiSCs has yet to be reported. The porting Table S1. derivation of iPS [28–30] and the potential to model and treat Oct4, the POU-family homeobox transcription factor, is human diseases using patient-derived iPS [31–34] has de- exclusively expressed in pluripotent cells of the developing emphasized the focus on using EpiSCs to understand human human and mouse embryo (inner cell mass [ICM] and early pluripotent stem cell-isms. Regardless, consistent differences germ cells), or its in vitro counterparts (ESCs and embryonic Schnerch, Cerdan, Bhatia 421 Table 1. Signaling pathways in mouse and human embryonic stem cells Expression Pathway Gene Human ESCs Mouse ESCs In Vivo/In Vitro Embryonic
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