Abstract
Somatic cells can be reprogrammed into induced pluripotent stem cells (iPSCs) through epigenetic manipulation. While the essential role of miRNA in reprogramming and maintaining pluripotency is well studied, little is known about the functions of miRNA from exosomes in this context. To fill this research gap,we comprehensively obtained the 17 sets of cellular mRNA transcriptomic data with 3.93 × 1010 bp raw reads and 18 sets of exosomal miRNA transcriptomic data with 2.83 × 107 bp raw reads from three categories of human somatic cells: peripheral blood mononuclear cells (PBMCs), skin fibroblasts(SFs) and urine cells (UCs), along with their derived iPSCs. Additionally, differentially expressed molecules of each category were identified and used to perform gene set enrichment analysis. Our study provides sets of comparative transcriptomic data of cellular mRNA and exosomal miRNA from three categories of human tissue with three individual biological controls in studies of iPSCs generation, which will contribute to a better understanding of donor cell variation in functional epigenetic regulation and differentiation bias in iPSCs.
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Background & Summary
Somatic reprogramming is a common method to manipulate cell lineage through epigenetic modification and induce somatic cells into a close embryonic stage triggered by various pluripotency master transcription factors, such as OCT4, SOX2, KLF4, c-Myc, or NANOG1,2,3,4. This somatic epigenetic manipulation has resulted in the generation of personalised induced pluripotent stem cells (iPSCs), which provide tremendous implications in regenerative medicine. During iPSCs generation, epigenetic regulation leads to differential patterns of gene expression through alterations in chromatin structure and modifications of the DNA while still sharing the same genomic sequence as its somatic cells5,6,7. Moreover, iPSCs retain epigenetic marks from their somatic source, known as “epigenetic memory”, which affects their downstream differentiation ability and inclines the differentiation to their original source8,9,10,11.
MiRNAs, a group of small non-coding RNAs with ~22-nt in length, were reported with solid evidence to maintain or manipulate cell lineage, which may be attributed to their ability to control factors involved in cell fate determination or epigenetic regulation12,13,14,15,16. For instance, miR-302 has been identified as a well-known gene silencer in reprogramming somatic cells into iPSCs. The miRNA function induces global DNA demethylation by repressing the expression of multiple key epigenetic regulators, such as DNMT1, MECP1/2, and HDAC2/412,17,18. The miR-290 family, called embryonic stem (ES) cell-specific cell cycle regulating miRNAs, was validated to maintain the rapid proliferative state of ES cells by regulating the G1-S phase transition. Moreover, miR-9 and miR-124a, which are predominantly expressed in neurons, have been demonstrated to regulate the formation and proliferation of the neural lineage derived from ES cells based on control of STAT3 phosphorylation19. Interestingly, miRNAs can be regulated by various epigenetic modifications, including DNA methylation, RNA modification, and histone modifications, which further exerts extensive influence on gene expression profile20,21,22,23,24. Dysregulation of the miRNA-epigenetic feedback loop has been validated to interfere with the physiological and pathological processes, but its specific role in cell fate determination of iPSCs remains poor understood.
Exosomes, one of the smallest extracellular vesicles (EVs) secreted in various cell types, act as bioactive vesicles in cell-to-cell communication by carrying proteins, miRNAs and other factors25,26. In the cell microenvironment, exosomal miRNA can be taken up by neighbour cells or distant cells and subsequently regulate the epigenetics of recipient cells. It was reported that exosomal miRNAs contribute significantly to the maintenance of pluripotency or other specific cell fate in their niche27,28,29,30. Notably, it was unveiled that about 70% of the miRNAs identified in iPSCs were also present in iPSC- EVs28, which indicates miRNAs were efficiently transferred from iPSCs to EVs for regulating pluripotent signalling. While the variate of exosomal miRNAs during reprogramming was limited to investigation, their roles in regulating cell fate need to be further studied.
To further understand the differentiation bias originating from somatic cells and the role of exosomal miRNA in regulating epigenetic heterogeneity during the generation of human iPSCs (hiPSCs), we simultaneously collected transcriptome data sets from the three most common somatic cell sources: skin fibroblasts (SFs), peripheral blood mononuclear cells (PBMCs), and urine cells (UCs), along with their derived iPSCs (Fig. 1). In order to minimise the biological variation, we recruited three healthy male donors within a similar age group (25–30 years old) and from the same genetic population (southern Han nationality represents about half of the Chinese population, approximately 10% of the world’s population31,32). Comparative data were generated before and after reprogramming, resulting in 17 sets of cellular RNA-Seq data and 18 sets of exosome-derived small RNA sequencing data. Subsequently, an in-house developed workflow was implemented to analyse the comparative transcriptomics data, including quality validation, differential expression analysis and gene set enrichment analysis. Our work provides a valuable resource for future investigations into donor cell variation in functional epigenetic regulation and differentiation bias in regenerative medicine.
Methods
Ethical approval
All samples were collected following the guidelines established by the Human Subject Research Ethics Committee at Guangzhou Institute of Biomedicine and Health (GIBH), the Chinese Academy of Sciences (CAS). The experiments were approved by the ethical committee under the approval number GIBH-IRB07-2015083. Prior to sample donation, all volunteers who donated skin, urine or blood samples had been thoroughly informed about the content, purposes, possible risks, and benefits of the experiment through a consent form, and provided their permission for genetic material data to be shared.
Collecting and culturing the human primary somatic cells
PBMCs, SFs and UCs were isolated from three healthy males aged from 25 to 30 and satisfied specific criteria. These criteria included having a normal BMI value, no family history of genetic disease and major surgery, and not smoking and alcohol consumption. Additionally, the annual health examination reports of all three volunteers had been thoroughly reviewed, indicating their physical well-being: no chronic illnesses or infectious diseases, with the standard range for blood pressure, heart rate, standard levels for blood chemistry, including cholesterol, glucose, liver function, kidney function, and absence of any medical conditions that significantly affect bodily functions or absence of diagnosed mental disorders. Notably, all three volunteers belonged to the southern Han nationality, representing approximately half of the Chinese national population and around 10% of the global population. The cell culture conditions were referred to in the previous publications33,34.
Establishment of hiPSCs from different sources
HiPSCs derived from SFs and UCs were generated based on the method described in the previous study33. The reprogramming procedure of UCs can be found in our previous protocol34. Reprogramming of human PBMCs was conducted with minor modification based on a published study35. Precisely, co-transfection of two episomal plasmids (pEP4-EO2SET2K and pEP4-M2L) and a vector containing hmiR302 cluster was performed in human PBMCs using AmaxaTM Basic NucleofectorTM Kit (Lonza), then these PBMCs were seeded on 6-well cell culture plate.
Characterisation of the hiPSCs
The karyotypes of hiPSCs derived from three types of somatic cell sources were detected using G band techniques. The presence of inserted genes from the reprogramming plasmids were demonstrated by PCR and gel-imaging system, as described in our previous research33. The protocols of immunofluorescence and quantitative real-time PCR analysis were referred to our previous studies33,34. Approximately 1 × 106 iPSCs were suspended in 100 μl Matrigel (diluted by DMEM/F12 1:1) and subcutaneously injected into the back of NOD/SCID mice. After teratoma formation, tumours were stained with haematoxylin-eosin and observed using an Olympus IX73 microscope.
RNA extraction, library construction, and Illumina sequencing
Total RNA was isolated using TRIzol reagent (Thermo Fisher) following its standard protocols. RNA qualification, library construction and sequencing were performed as our previous publications33,34.
Exosomal miRNA extraction, library construction, and Illumina sequencing
Exosomes were isolated from the cell culture medium using exoRNeasy Serum/Plasma Maxi Kit (Qiagen) following the provided instructions. HiPSCs with passage numbers ranging from 21 to 28 were used to isolate the exosomes in our experiments. Exosomal miRNA was extracted utilizing the exoRNeasy Mini Kit (Qiagen). After quantifying and qualifying the RNA, 3 μg of total RNA was used for the construction of sequencing libraries through NEBNext Multiplex Small RNA Library Prep Set for Illumina (NEB, USA). The quality of each library was assessed by the Agilent Bioanalyzer 2100 system, and sequencing was done on an Illumina Hiseq 2500/2000 platform.
Preprocessing of RNA sequencing data
Raw sequencing data were processed by fastp v0.20.136 with default parameters to remove adapter sequences, low-quality reads and short-length sequences. The resulting clean reads were then mapped to the human genome hg38 to quantify global gene expression using the expectation-Maximization method implemented in RSEM v1.2.22. Gene count and transcripts per million (TPM) matrix information was obtained for each sample. After filtering low-expression genes with an average expression less than 1, the log2(CPM + 1) values of each sample were used for the principal component analysis (PCA) and the correlation coefficient calculation. Differentially expressed genes (DEGs) between somatic cells and their derived hiPSCs were determined by DESeq 2 v1.30.137. DEGs were identified with an adjusted P-value < 0.05 and the absolute value of log2FoldChange > 1. While TPM matrix information was used to evaluate the expression levels of differentiation and pluripotency related genes. Gene set enrichment analysis was conducted by clusterProfiler v4.4.438 based on Gene Ontology (GO) and the Kyoto Encyclopedia of Genes and Genomes (KEGG) databases.
Preprocessing of small RNA sequencing data
FastQC (http://www.bioinformatics.babraham.ac.uk/projects/fastqc/) was utilized to verify the sequence quality by assessing parameters such as Q20, Q30, GC-content and adapter sequences. The raw data were mapped to the human genome hg38 and human miRNA sequences from miRBase39 v22 to predict novel miRNAs and evaluate the expression levels of known miRNAs by miRDeep40 v2.0.1.2 in multiple samples mode. The mapping number of each read in the human genome hg38 was constrained by a maximum of 5. Low-expression miRNAs with an average expression of less than 1 were filtered out, and then log2(CPM + 1) values of miRNA in each sample were used to perform the principal component analysis (PCA) and calculate the correlation coefficient. Differentially expressed miRNAs (DEMs) were identified using the same method as for DEGs. The miRNA targeted genes were analysed by multiMiR41 v1.12.0 based on its database v2.3, which incorporated three validated miRNA-target interactions databases (miRecoord, miRtarBase and TarBase) and 8 predicted miRNA-target interactions databases (DIANA-microT-CDS, RIMMo, MicroCosm, miRDB, PicTar, PITA and targetScan). The down-regulated DEGs were used to explore the up-regulated DEMs target genes, while the up-regulated DEGs were used to explore the down-regulated DEMs target genes. The targeted genes found in validated databases or in more than three predicted databases underwent GO and KEGG pathway enrichment, as described above.
Data Records
Our raw data, consisting of 17 RNA-seq and 18 exosomes’ small RNA-seq data sets, was stored in the Genome Sequence Archive42 in National Genomics Data Center (NGDC)43 with the accession number HRA00369744. The corresponding expression matrix information was deposited in NGDC of China National Center for Bioinformation with the accession number PRJCA01366245.
Technical Validation
The characteristics of hiPSCs
Karyotype analysis indicated that the chromosome profiles of hiPSCs derived from PBMCs, SFs and UCs were without abnormality (Fig. 2a). The exogenous episomal DNA (OCT4, SOX2, KLF4, MicroRNA302-367 or c-Myc) was absent in all three somatic cell-derived hiPSCs (Fig. 2b). Compared with their respective somatic cells, the mRNA expression levels of OCT4, SOX2 and NANOG were significantly increased in hiPSCs from three categories (Fig. 2c). Immunofluorescence result revealed higher expression levels of the pluripotent protein markers (OCT4, SSEA4, TRA-1-60, and TRA-1-81) in hiPSCs derived from the three somatic cell types compared to their respective somatic cells (Fig. 2d). The pluripotent potential of hiPSCs derived from the three types of somatic cells was further investigated by teratoma formation, which exhibited their ability to differentiate into three germ-layer (Fig. 2e). These results illustrated that our hiPSCs have similar pluripotent characteristics to human embryonic stem cells.
Quality control of RNA sequencing data
High-throughput RNA sequencing generated 1.2~2.0 × 107 raw reads per sample, with the Q20 > 0.95, Q30 > 0.90, GC-content close to 0.50, and the mean length of 149 bp for clean reads(Table 1). Most of the pair reads were aligned to the hg38 genome, with the sample align rate ranging from 66% to 88% and the unique mapping rate ranging from 15% to 22%, much lower than that of the multiple mapping rate (Table 2).
Genes expression analysis
Gene expression analysis was performed using three biologically replicated samples, except that SFs-derived hiPSCs had only two samples. The correlation coefficients calculated based on Pearson correlation and PCA analysis implied good repeatability within the biological replicates and high similarity among hiPSCs derived from the three kinds of somatic cells (Fig. 3a,b). The gene expression levels of the pluripotency related genes (PPA2, GDF3, TERT, NANOG, ZFP42, FGF4, LIN28A, DPPA4, POU5F1, and SOX2) were significantly higher in hiPSCs than that in somatic cells, whereas the gene expression levels of differentiation related genes (NR2F2, ANPEP, and SOX17) were significantly lower (Fig. 3c). The analysis of differentially expressed genes (DEGs) between somatic cells and their derived hiPSCs indicated significant changes from a differentiated state to a pluripotency state. In these sets, a total of 10,203 genes, 10,026 genes and 9,330 genes were identified as DEGs (Table 3), respectively, including 4446 shared DEGs with 2279 consistently up-regulated DEGs and 1423 consistently down-regulated DEGs. (Fig. 3d). In addition, the top 20 pathways identified in the enrichment analysis of DEGs based on the biological process GO and KEGG indicated that immune-related pathways were mostly down-regulated in PBMCs-derived hiPSCs compared to PBMCs. Up-regulated synaptic signalling and down-regulated embryonic skeletal system development were both observed in the SFs and UCs categories (Fig. 3e,f).
Processing the small RNA sequencing data
A total of 2.83 × 107 raw reads were generated, ranging from 1.1 to 2.2 × 106 raw reads per sample. The Q20 rate was more than 0.99, the Q30 rate was more than 0.98, and the GC content of raw reads with a mean length of 50 bp was approximately 0.53 (Table 4). In summary, the small RNA-seq sequencing data have quite good quality. The mapping rate of small RNA reads to the hg38 genome per sample ranged from 19% to 62%, while the mapping rate of reads to human miRNA sequences ranged from 0.5% to 12%, with an average rate of 2.50% (Table 5). According to data GSE21655646 deposited on the NCBI GEO database, the mapping rates of small RNA sequences extracted from exosomes to human miRNA sequences ranged from 1.26% to 2.68%, indicating a lower alignment rate of miRNA from exosomes compared to the cellular datasets. The results of correlation coefficients and the PCA analysis implied good repeatability within the biological replicates, except for sample Us3, which displays a closer relationship to the PBMCs exosomes (Fig. 4a,b). The exosomal miRNA data generated from three categories of hiPSCs exhibited a high degree of similarity, vastly different from their somatic miRNA samples. A total of 248, 137 and 106 miRNAs were separately identified as differentially expressed miRNAs (DEMs) in these three groups, including shared 72 DEMs with 68 consistently up-regulated DEMs and 5 consistently down-regulated DEMs (Table 6, Figs. 3d, 4c). The items of DEGs targeted by DEMs were summarised in Table 7. The gene set enrichment analysis of targeted DEGs for each group based on biological process GO, and KEGG was conducted and revealed similar enriched pathways to DEGs (Table 8). The top 20 enriched pathways were shown in Fig. 4e,f.
Code availability
The command script for MirDeep2 and downstream analysis code written by R are available at the GitHub repository https://github.com/Andelyu/hiPSCs_exosomal_miRNA_project.
References
Ding, Y. et al. OCT4, SOX2 and NANOG co-regulate glycolysis and participate in somatic induced reprogramming. Cytotechnology 74, 371–383, https://doi.org/10.1007/s10616-022-00530-6 (2022).
Takahashi, K. & Yamanaka, S. Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors. Cell 126, 663–676, https://doi.org/10.1016/j.cell.2006.07.024 (2006).
Wernig, M. et al. In vitro reprogramming of fibroblasts into a pluripotent ES-cell-like state. Nature 448, 318–324, https://doi.org/10.1038/nature05944 (2007).
Gomes, K. M. et al. Induced pluripotent stem cells reprogramming: Epigenetics and applications in the regenerative medicine. Revista da Associacao Medica Brasileira (1992) 63, 180–189, https://doi.org/10.1590/1806-9282.63.02.180 (2017).
Wang, Y., Bi, Y. & Gao, S. Epigenetic regulation of somatic cell reprogramming. Current Opinion in Genetics & Development 46, 156–163, https://doi.org/10.1016/j.gde.2017.07.002 (2017).
Villasante, A. et al. Epigenetic regulation of Nanog expression by Ezh2 in pluripotent stem cells. Cell cycle (Georgetown, Tex.) 10, 1488–1498, https://doi.org/10.4161/cc.10.9.15658 (2011).
Watanabe, A., Yamada, Y. & Yamanaka, S. Epigenetic regulation in pluripotent stem cells: a key to breaking the epigenetic barrier. Philosophical transactions of the Royal Society of London. Series B, Biological sciences 368, 20120292, https://doi.org/10.1098/rstb.2012.0292 (2013).
Nashun, B., Hill, P. W. & Hajkova, P. Reprogramming of cell fate: epigenetic memory and the erasure of memories past. The EMBO journal 34, 1296–1308, https://doi.org/10.15252/embj.201490649 (2015).
Kim, M. H., Thanuthanakhun, N., Fujimoto, S. & Kino-Oka, M. Effect of initial seeding density on cell behavior-driven epigenetic memory and preferential lineage differentiation of human iPSCs. Stem cell research 56, 102534, https://doi.org/10.1016/j.scr.2021.102534 (2021).
Wu, T. et al. Histone Variant H2A.X Deposition Pattern Serves as a Functional Epigenetic Mark for Distinguishing the Developmental Potentials of iPSCs. Cell Stem Cell 15, 281–294, https://doi.org/10.1016/j.stem.2014.06.004 (2014).
Rouhani, F. et al. Genetic background drives transcriptional variation in human induced pluripotent stem cells. PLoS genetics 10, e1004432, https://doi.org/10.1371/journal.pgen.1004432 (2014).
Ying, S. Y., Fang, W. & Lin, S. L. The miR-302-Mediated Induction of Pluripotent Stem Cells (iPSC): Multiple Synergistic Reprogramming Mechanisms. Methods in molecular biology (Clifton, N.J.) 1733, 283–304, https://doi.org/10.1007/978-1-4939-7601-0_23 (2018).
Giacomazzi, G. et al. MicroRNAs promote skeletal muscle differentiation of mesodermal iPSC-derived progenitors. Nature communications 8, 1249, https://doi.org/10.1038/s41467-017-01359-w (2017).
Ivey, K. N. et al. MicroRNA regulation of cell lineages in mouse and human embryonic stem cells. Cell Stem Cell 2, 219–229, https://doi.org/10.1016/j.stem.2008.01.016 (2008).
Izarra, A. et al. miRNA-1 and miRNA-133a are involved in early commitment of pluripotent stem cells and demonstrate antagonistic roles in the regulation of cardiac differentiation. Journal of tissue engineering and regenerative medicine 11, 787–799, https://doi.org/10.1002/term.1977 (2017).
Liu, Y. et al. MicroRNA-200a regulates Grb2 and suppresses differentiation of mouse embryonic stem cells into endoderm and mesoderm. PloS one 8, e68990, https://doi.org/10.1371/journal.pone.0068990 (2013).
Lin, S. L. & Ying, S. Y. Mechanism and Method for Generating Tumor-Free iPS Cells Using Intronic MicroRNA miR-302 Induction. Methods in molecular biology (Clifton, N.J.) 1733, 265–282, https://doi.org/10.1007/978-1-4939-7601-0_22 (2018).
Lin, S. L., Chen, J. S. & Ying, S. Y. MiR-302-Mediated Somatic Cell Reprogramming and Method for Generating Tumor-Free iPS Cells Using miR-302. Methods in molecular biology (Clifton, N.J.) 2115, 199–219, https://doi.org/10.1007/978-1-0716-0290-4_12 (2020).
Delaloy, C. et al. MicroRNA-9 coordinates proliferation and migration of human embryonic stem cell-derived neural progenitors. Cell Stem Cell 6, 323–335, https://doi.org/10.1016/j.stem.2010.02.015 (2010).
Szulwach, K. E. et al. Cross talk between microRNA and epigenetic regulation in adult neurogenesis. The Journal of cell biology 189, 127–141, https://doi.org/10.1083/jcb.200908151 (2010).
Au, S. L. et al. Enhancer of zeste homolog 2 epigenetically silences multiple tumor suppressor microRNAs to promote liver cancer metastasis. Hepatology (Baltimore, Md.) 56, 622–631, https://doi.org/10.1002/hep.25679 (2012).
Dastmalchi, N. et al. An Updated Review of the Cross-talk Between MicroRNAs and Epigenetic Factors in Cancers. Current medicinal chemistry 28, 8722–8732, https://doi.org/10.2174/0929867328666210514125955 (2021).
Xu, N., Liu, J. & Li, X. Lupus nephritis: The regulatory interplay between epigenetic and MicroRNAs. Frontiers in physiology 13, 925416, https://doi.org/10.3389/fphys.2022.925416 (2022).
Guil, S. & Esteller, M. DNA methylomes, histone codes and miRNAs: tying it all together. The international journal of biochemistry & cell biology 41, 87–95, https://doi.org/10.1016/j.biocel.2008.09.005 (2009).
Bang, C. & Thum, T. Exosomes: New players in cell–cell communication. The international journal of biochemistry & cell biology 44, 2060–2064, https://doi.org/10.1016/j.biocel.2012.08.007 (2012).
Kalluri, R. & LeBleu, V. S. The biology, function, and biomedical applications of exosomes. 367, https://doi.org/10.1126/science.aau6977 (2020).
La Greca, A. et al. Extracellular vesicles from pluripotent stem cell-derived mesenchymal stem cells acquire a stromal modulatory proteomic pattern during differentiation. Experimental & Molecular Medicine 50, 1–12, https://doi.org/10.1038/s12276-018-0142-x (2018).
Baumann, K. EVs promote stemness. Nature Reviews Molecular Cell Biology 22, 72–73, https://doi.org/10.1038/s41580-020-00327-5 (2021).
Bi, Y. et al. Systemic proteomics and miRNA profile analysis of exosomes derived from human pluripotent stem cells. Stem Cell Research & Therapy 13, 449, https://doi.org/10.1186/s13287-022-03142-1 (2022).
Jung, J. H., Fu, X. & Yang, P. C. Exosomes Generated From iPSC-Derivatives: New Direction for Stem Cell Therapy in Human Heart Diseases. Circulation research 120, 407–417, https://doi.org/10.1161/circresaha.116.309307 (2017).
Cong, P. K., Bai, W. Y. & Li, J. C. Genomic analyses of 10,376 individuals in the Westlake BioBank for Chinese (WBBC) pilot project. 13, 2939, https://doi.org/10.1038/s41467-022-30526-x (2022).
Xu, S. et al. Genomic dissection of population substructure of Han Chinese and its implication in association studies. American journal of human genetics 85, 762–774, https://doi.org/10.1016/j.ajhg.2009.10.015 (2009).
Fan, K. et al. A Machine Learning Assisted, Label-free, Non-invasive Approach for Somatic Reprogramming in Induced Pluripotent Stem Cell Colony Formation Detection and Prediction. Scientific reports 7, 13496, https://doi.org/10.1038/s41598-017-13680-x (2017).
Sun, W. et al. Human Urinal Cell Reprogramming: Synthetic 3D Peptide Hydrogels Enhance Induced Pluripotent Stem Cell Population Homogeneity. 6, 6263–6275, https://doi.org/10.1021/acsbiomaterials.0c00667 (2020).
Staerk, J. et al. Reprogramming of human peripheral blood cells to induced pluripotent stem cells. Cell Stem Cell 7, 20–24, https://doi.org/10.1016/j.stem.2010.06.002 (2010).
Chen, S., Zhou, Y., Chen, Y. & Gu, J. fastp: an ultra-fast all-in-one FASTQ preprocessor. Bioinformatics (Oxford, England) 34, i884–i890, https://doi.org/10.1093/bioinformatics/bty560 (2018).
Love, M. I., Huber, W. & Anders, S. Moderated estimation of fold change and dispersion for RNA-seq data with DESeq 2. Genome biology 15, 550, https://doi.org/10.1186/s13059-014-0550-8 (2014).
Wu, T. et al. clusterProfiler 4.0: A universal enrichment tool for interpreting omics data. Innovation (Cambridge (Mass.)) 2, 100141, https://doi.org/10.1016/j.xinn.2021.100141 (2021).
Kozomara, A., Birgaoanu, M. & Griffiths-Jones, S. miRBase: from microRNA sequences to function. Nucleic acids research 47, D155–d162, https://doi.org/10.1093/nar/gky1141 (2019).
Friedländer, M. R., Mackowiak, S. D., Li, N., Chen, W. & Rajewsky, N. miRDeep2 accurately identifies known and hundreds of novel microRNA genes in seven animal clades. Nucleic acids research 40, 37–52, https://doi.org/10.1093/nar/gkr688 (2012).
Ru, Y. et al. The multiMiR R package and database: integration of microRNA-target interactions along with their disease and drug associations. Nucleic acids research 42, e133, https://doi.org/10.1093/nar/gku631 (2014).
Chen, T. et al. The Genome Sequence Archive Family: Toward Explosive Data Growth and Diverse Data Types. Genomics, proteomics & bioinformatics https://doi.org/10.1016/j.gpb.2021.08.001 (2021).
Database Resources of the National Genomics Data Center, China National Center for Bioinformation in 2022. Nucleic acids research 50, D27–d38, https://doi.org/10.1093/nar/gkab951 (2022).
National Genomics Data Center-GSA for Human https://ngdc.cncb.ac.cn/gsa-human/browse/HRA003697 (2023).
Genome Sequence Archive in National Genomics Data Center https://ngdc.cncb.ac.cn/bioproject/browse/PRJCA013662 (2023).
NCBI Gene Expression Omnibus https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE216556 (2022).
Acknowledgements
This research was funded by multiple sources, including the Key Research Program of Chinese Academy of Sciences (O2222001), the Scientific Instrumentation Development Program of Chinese Academy of Sciences (ZDKYYQ20210006), the National Natural Science Foundation (No. 62271127), and the Medico-Engineering Cooperation Funds from the University of Electronic Science and Technology of China and West China Hospital of Sichuan University (No. ZYGX2022YGRH011 and HXDZ22005).
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Xiao Zhang and Sheng Zhang conceived and designed the experiments. Xiao Zhang, Chunlai Yu, Sheng Zhang and Nini Rao wrote and revised the paper. Shaoling Wu collected the specimens. Mei Zhang took the charge of all experiments. Chunlai Yu completed the analysis work. Yucui Xiong, Qizheng Wang, Yuanhua Wang, Zhizhong Zhang and Yan Wang assisted with the experiments, Sajjad Hussain assisted in revising the language of the paper.
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Yu, C., Zhang, M., Xiong, Y. et al. Comparison of miRNA transcriptome of exosomes in three categories of somatic cells with derived iPSCs. Sci Data 10, 616 (2023). https://doi.org/10.1038/s41597-023-02493-5
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DOI: https://doi.org/10.1038/s41597-023-02493-5