Several studies have reported pivotal

Several studies have reported pivotal roles for some miRNAs in hBMSC-lineage commitment and fate determination that are mediated by their ability to modulate differentiation-regulating factors. For instance, miR-210, miR-206, miR-133, miR-2861, and miR-138 have been implicated in the regulation of osteogenic differentiation via modulation of the TGFB type I receptor, connexin 43, RUNX2, HDACs, and the FAK, while miR-21, miR-375, miR-27a, and miR-320c have been involved in the regulation of adipogenic differentiation through modulation of TGFB1, ERK1/2, PPARG, and RUNX2 (Eskildsen et al., 2011; Hamam et al., 2014; Hu et al., 2011; Inose et al., 2009; Kim et al., 2010b, 2009; Li et al., 2008; Ling et al., 2011; Mizuno et al., 2009). To identify the molecular mechanism regulating LRP3 tegaserod in hBMSCs, we performed miRNA expression profiling in the imCL1 and imCL2 clones, which revealed some differentially expressed miRNAs between the two clones. Interestingly, hsa-miR-4739, which we subsequently identified as a bone fide regulator of LRP3 expression, was the most under-represented miRNA in imCL1 compared to imCL2. Thus far, only three studies have reported on hsa-miR-4739 functions, linking it to unilineage hematopoietic stem cell differentiation and acute myeloid leukemia, β-catenin signaling pathway and gastric cancer, as well as in the pathogenesis of negative anaplastic large cell lymphoma (Cattaneo et al., 2015; Dong et al., 2015; Wang et al., 2015a). Hence, our data extend the list of possible biological roles of hsa-miR-4739 by demonstrating its ability to regulate LRP3-mediated osteogenic at the expense of adipogenic differentiation of hBMSCs, which suggests that it functions as a molecular switch during hSSC differentiation. It is plausible that hsa-miR-4739 has additional gene targets besides LRP3 that might also regulate hSSC/BMSC osteogenic and adipocytic differentiation. In fact, investigation of the TargetScan database identified LRP4 as a potential target for hsa-miR-4739; however, the direct regulation of LRP4 by hsa-miR-4739 remains to be proven. Therefore, we propose the use of hsa-miR-4739 mimics or inhibitors to fine-tune the commitment of hBMSCs to the osteogenic or the adipogenic lineage, with potential application in regenerative medicine. The following are the supplementary data related to this article.
Conflict of interest
Acknowledgements The authors would like to extend their sincere appreciation to the Deanship of Scientific Research at King Saud University for its funding of this research through the Research Group Project no RGP-1438-032.
: please write over examples provided, fill in all fields. Resource details Bernard-Soulier Syndrome (BSS) is an autosomal recessive rare bleeding disorder characterized by thrombocytopenia and large platelets. The disease is caused by mutations in one of the three genes coding the glycoprotein complex GPIb-IX-V, which is a receptor for von Willebrand Factor (vWF). As a result, the complex is absent or dysfunctional in the cell membrane of BSS platelets (Andrews and Berndt, 2013; Berndt and Andrews, 2011). We have recently described the first two human iPSC models for BSS (Lopez-Onieva et al., 2016a; Lopez-Onieva et al., 2016b). In this study, we report the generation of a new cellular model from a BSS patient carrying the homozygous point mutation c.212 T>C in the GPIX locus (Noris et al., 1997). Human Peripheral Blood Mononuclear Cells (PBMC) from this patient (BSS3-PBMC) were reprogrammed into iPSCs (BSS3-PBMC-iPS4F8) using the non-integrative CytoTune iPS 2.0 Reprograming System (Life Technologies, Invitrogen). Characterization of BSS3-PBMC-iPS4F8 line involved different molecular and functional analysis. First, sequencing analysis of the GPIX gene confirmed the presence of a c.212 T>C transition in exon 3 predicting a homozygous p.Phe55Ser substitution, identical to PBMCs-BSS3 (Fig. 1A). In addition, Short Tandem Repeat (STR) profiling confirmed same genetic identity between both cell types (Fig. 1B). Moreover, exogenous reprogramming factors were silenced after 9 passages (Fig. 1C). PCR-based detection tests confirmed the absence of Mycoplasma contamination at this stage (data not shown). Importantly, BSS3-PBMC-iPS4F8 line showed normal karyotype (46, XX) (Fig. 1D).