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Material andmethods
Results anddiscussion
Conclusions Here we report a comprehensive and unique transcriptomic study of the kinetics of gene SM-164 during ASC adipogenesis in vitro. Despite the limitation of the study design including the use of relatively late passage ASCs and the lack of BMI measurements for all four donors, the study nonetheless identified genes that could potentially be used to distinguish adipocytes at the early (SLC7A8 and TMEM158) and late (COL4A1, PCK1, RORB, CEBPA and PPARG) stages of differentiation, while others (FABP4, PLIN1, ADIPOQ, AOC3, IGF1, ACSL5 and APCDD1) were identified to be general adipocyte markers. ASCs were also found to express unique gene signatures on different days during in vitro adipogenesis. The identification of FOXO1 and its functional interacting partners such as SIRT1, whose role in adipogenesis has been well described in US patent US20050136429 A1 (Guarente and Picard, 2005), validates our hypothesis and the methodology that could be used to identify potential anti-obesity therapeutic targets. The identification of MED1, EP300 and NCOA1 as functional partners interacting strongly with the anti-diabetic thiazolidinedione (TZD) drug target PPARG (Lehrke and Lazar, 2005) warrants further investigation in vivo to establish their suitability as potential drug targets in combating obesity and its associated diseases. PER1 and RORB are two important circadian clock genes identified in this study, which could play a role in controlling the internal circadian clock driving adipogenesis/adipocyte formation. MLXIPL and ZNF117 TFs, which have not previously been reported to play a role in adipogenesis, were identified to be significantly up-regulated from D14 onwards. Functional studies of these TFs will provide greater insight into their role in adipogenesis and might serve as potential targets for regulating this process. Functional network analysis revealed that ASC adipogenesis leads to significant enrichment of genes associated with different pathophysiological conditions including various cancers and cardiovascular signaling pathways. For example, adipocyte maturation correlates with the activation of colorectal cancer metastasis signaling, strengthening the link between obesity and the risk of cancer development. Overall, this study provides a unique resource for many functional studies of genes with no previously reported role in adipogenesis. The following are the supplementary data related to this article.
Competing interests
Authors\' contributions
Acknowledgements This research was funded by the South African Medical Research Council in terms of the SAMRC\'s Flagship Award Project SAMRC-RFA-UFSP-01-2013/STEM CELLS, the SAMRC Extramural Stem Cell Research and Therapy Unit, the National Research Foundation of South Africa (grant no. 86942), the National Health Laboratory Services Research Trust (grant no. 94453) and the Institute for Cellular and Molecular Medicine of the University of Pretoria.
Introduction Human pluripotent stem cell (hPSC)-derived cardiomyocytes (CMs) have provided a realistic and exciting platform for drug-induced toxicity screening and disease modeling (Burridge et al., 2012; Yang et al., 2014). Directed differentiation of CMs from hPSCs has been established in several laboratories (reviewed in (Burridge et al., 2012; Mummery et al., 2012)), producing immature CMs that have limited ability to accurately reflect the physiology and pathology of human hearts (Nunes et al., 2013; Yang et al., 2014). Evidence suggests hPSC-CMs can progressively mature in long-term culture (Kamakura et al., 2013; Lundy et al., 2013), but this method can be time consuming which may limit its utility. Thus, new approaches that enhance hPSC-CM maturation are needed. In adult myocardium, CMs are longitudinally aligned in the form of parallel bundles to facilitate the mechanical contraction and electrical propagation of the heart tissue. Mimicking nature\'s work, engineered anisotropy (namely cell patterning) has been widely used as an effective approach to improve the maturation of immature CMs (Yang et al., 2014). This engineered anisotropy can be induced by electrical (Nunes et al., 2013; Radisic et al., 2004) or mechanical loading (Thavandiran et al., 2013; Tulloch et al., 2011) and also by substrates that provide topographical guidance, which create a permissive environment to guide the alignment of CMs (Chen et al., 2014; Kim et al., 2010; Liau et al., 2011; Ma et al., 2014; McCain et al., 2014; Rao et al., 2013; Salick et al., 2014; Zhang et al., 2013). Aligned CMs may result in enhanced cardiac phenotypes indicative of cell maturation. Recently, enhanced sarcomere alignment (Khan et al., 2015; Salick et al., 2014), excitation-contraction coupling (Zhang et al., 2013), calcium cycling (Khan et al., 2015; Rao et al., 2013), and drug sensitivity (Chen et al., 2014) have been reported for hPSC-CMs grown on various micro-patterned substrates, suggesting improved cell structural and/or functional maturation. However, the robustness of cell alignment as a single approach to improve the maturation of hPSC-CMs has not been fully investigated.