Neurological Disorders Research Center, Qatar Biomedical Research Institute, Hamad Bin Khalifa University, Qatar Foundation, Doha P.O. Box 34110, Qatar.
Qatar Biomedical Research Institute, Hamad Bin Khalifa University, Qatar Foundation, Doha P.O. Box 34110, Qatar.
Cells. 2022 Nov 29;11(23):3833. doi: 10.3390/cells11233833.
Somatic cells are reprogrammed with reprogramming factors to generate induced pluripotent stem cells (iPSCs), offering a promising future for disease modeling and treatment by overcoming the limitations of embryonic stem cells. However, this process remains inefficient since only a small percentage of transfected cells can undergo full reprogramming. Introducing miRNAs, such as miR-294 and miR302/3667, with reprogramming factors, has shown to increase iPSC colony formation. Previously, we identified five transcription factors, GBX2, NANOGP8, SP8, PEG3, and ZIC1, which may boost iPSC generation. In this study, we performed quantitative miRNAome and small RNA-seq sequencing and applied our previously identified transcriptome to identify the potential miRNA-mRNA regulomics and regulatory network of other ncRNAs. From each fibroblast (N = 4), three iPSC clones were examined (N = 12). iPSCs and original fibroblasts expressed miRNA clusters differently and miRNA clusters were compared to mRNA hits. Moreover, miRNA, piRNA, and snoRNAs expression profiles in iPSCs and original fibroblasts were assessed to identify the potential role of ncRNAs in enhancing iPSC generation, pluripotency, and differentiation. Decreased levels of let-7a-5p showed an increase of SP8 as described previously. Remarkably, the targets of identifier miRNAs were grouped into pluripotency canonical pathways, on stemness, cellular development, growth and proliferation, cellular assembly, and organization of iPSCs.
体细胞被重编程因子重编程为诱导多能干细胞（iPSCs），通过克服胚胎干细胞的局限性，为疾病建模和治疗提供了有前途的未来。然而，这个过程仍然效率低下，因为只有一小部分转染细胞可以进行完全重编程。引入重编程因子，如 miR-294 和 miR302/3667，可以增加 iPSC 集落形成。以前，我们鉴定了五个转录因子，GBX2、NANOGP8、SP8、PEG3 和 ZIC1，它们可能会促进 iPSC 的生成。在这项研究中，我们进行了定量 miRNA 组学和小 RNA-seq 测序，并应用我们之前鉴定的转录组来鉴定其他 ncRNAs 的潜在 miRNA-mRNA 调控组学和调控网络。从每个成纤维细胞（N=4）中，检查了三个 iPSC 克隆（N=12）。iPSCs 和原始成纤维细胞表达的 miRNA 簇不同，并且将 miRNA 簇与 mRNA 命中进行了比较。此外，还评估了 iPSCs 和原始成纤维细胞中 miRNA、piRNA 和 snoRNA 的表达谱，以确定 ncRNAs 在增强 iPSC 生成、多能性和分化中的潜在作用。如前所述，let-7a-5p 的水平降低导致 SP8 增加。值得注意的是，鉴定 miRNA 的靶标被分为多能性经典途径、干性、细胞发育、生长和增殖、细胞组装和 iPSC 的组织。
