# Impact of microRNA-130a on the neutrophil proteome

**Authors:** Corinna Cavan Pedersen, Jan Christian Refsgaard, Ole Østergaard, Lars Juhl Jensen, Niels Henrik Helweg Heegaard, Niels Borregaard, Jack Bernard Cowland

PMC · DOI: 10.1186/s12865-015-0134-8 · BMC Immunology · 2015-11-25

## TL;DR

This study explores how microRNA-130a affects the protein levels in neutrophils, revealing new insights into its role in neutrophil development.

## Contribution

The study experimentally identifies miR-130a-regulated proteins and constructs an association network of potential targets.

## Key findings

- Inhibition of miR-130a significantly regulated 44 and 34 proteins in murine and human myeloid cell lines.
- Three proteins (NFYC, ISOC1, and CAT) were identified as putative miR-130a targets with high RAIN scores.
- Transcription factors Myb and CBF-β were linked as putative miR-130a targets, potentially regulating granule proteins.

## Abstract

MicroRNAs (miRNAs) are important for the development and function of neutrophils. miR-130a is highly expressed during early neutrophil development and regulates target proteins important for this process. miRNA targets are often identified by validating putative targets found by in silico prediction algorithms one at a time. However, one miRNA can have many different targets, which may vary depending on the context. Here, we investigated the effect of miR-130a on the proteome of a murine and a human myeloid cell line.

Using pulsed stable isotope labelling of amino acids in cell culture and mass spectrometry for protein identification and quantitation, we found 44 and 34 proteins that were significantly regulated following inhibition of miR-130a in a miR-130a-overexpressing 32Dcl3 clone and Kasumi-1 cells, respectively. The level of miR-130a inhibition correlated with the impact on protein levels. We used RAIN, a novel database for miRNA–protein and protein–protein interactions, to identify putative miR-130a targets. In the 32Dcl3 clone, putative targets were more up-regulated than the remaining quantified proteins following miR-130a inhibition, and three significantly derepressed proteins (NFYC, ISOC1, and CAT) are putative miR-130a targets with good RAIN scores. We also created a network including inferred, putative neutrophil miR-130a targets and identified the transcription factors Myb and CBF-β as putative miR-130a targets, which may regulate the primary granule proteins MPO and PRTN3 and other proteins differentially expressed following miR-130a inhibition in the 32Dcl3 clone.

We have experimentally identified miR-130a-regulated proteins within the neutrophil proteome. Linking these to putative miR-130a targets, we provide an association network of potential direct and indirect miR-130a targets that expands our knowledge on the role of miR-130a in neutrophil development and is a valuable platform for further experimental studies.

The online version of this article (doi:10.1186/s12865-015-0134-8) contains supplementary material, which is available to authorized users.

## Linked entities

- **Genes:** MIR130A (microRNA 130a) [NCBI Gene 406919], NFYC (nuclear transcription factor Y subunit gamma) [NCBI Gene 4802], ISOC1 (isochorismatase domain containing 1) [NCBI Gene 51015], CAT (catalase) [NCBI Gene 847], MYB (MYB proto-oncogene, transcription factor) [NCBI Gene 4602], CBFB (core-binding factor subunit beta) [NCBI Gene 865], MPO (myeloperoxidase) [NCBI Gene 4353], PRTN3 (proteinase 3) [NCBI Gene 5657]
- **Species:** Mus musculus (taxon 10090)

## Full-text entities

- **Genes:** Smad4 (SMAD family member 4) [NCBI Gene 17128] {aka D18Wsu70e, DPC4, Madh4}, Prtn3 (proteinase 3) [NCBI Gene 19152] {aka PR-3, PR3, mPR3}, Il2ra (interleukin 2 receptor, alpha chain) [NCBI Gene 16184] {aka CD25, Il2r, Ly-43}, PRTN3 (proteinase 3) [NCBI Gene 5657] {aka ACPA, AGP7, C-ANCA, CANCA, MBN, MBT}, ISOC1 (isochorismatase domain containing 1) [NCBI Gene 51015] {aka CGI-111}, Tgfb1 (transforming growth factor, beta 1) [NCBI Gene 21803] {aka TGF-beta1, TGFbeta1, Tgfb, Tgfb-1}, IL3 (interleukin 3) [NCBI Gene 3562] {aka IL-3, MCGF, MULTI-CSF}, RASIP1 (Ras interacting protein 1) [NCBI Gene 54922] {aka RAIN}, SACM1L (SAC1 like phosphatidylinositide phosphatase) [NCBI Gene 22908] {aka SAC1}, Mir130a (microRNA 130a) [NCBI Gene 387149] {aka Mirn130, Mirn130a, mir-130a}, NFYC (nuclear transcription factor Y subunit gamma) [NCBI Gene 4802] {aka CBF-C, CBFC, H1TF2A, HAP5, HSM, NF-YC}, CAT (catalase) [NCBI Gene 847], SMAD4 (SMAD family member 4) [NCBI Gene 4089] {aka DPC4, JIP, MADH4, MYHRS}, RUNX1 (RUNX family transcription factor 1) [NCBI Gene 861] {aka AML1, AML1-EVI-1, AMLCR1, CBF2alpha, CBFA2, EVI-1}, Rasip1 (Ras interacting protein 1) [NCBI Gene 69903] {aka 1110025D03Rik, 2610025P08Rik, Rain}, Cat (catalase) [NCBI Gene 12359] {aka 2210418N07, Cas-1, Cas1, Cs-1}, CEBPE (CCAAT enhancer binding protein epsilon) [NCBI Gene 1053] {aka C/EBP-epsilon, CRP1, IMD108, SGD1, c/EBP epsilon}, Isoc1 (isochorismatase domain containing 1) [NCBI Gene 66307] {aka 2610034N03Rik}, Nfya (nuclear transcription factor-Y alpha) [NCBI Gene 18044] {aka Cbf-b, SEZ-10, SEZ10}, CEBPA (CCAAT enhancer binding protein alpha) [NCBI Gene 1050] {aka C/EBP-alpha, CEBP}, CBFB (core-binding factor subunit beta) [NCBI Gene 865] {aka CLCD2, PEBP2B}, NFYA (nuclear transcription factor Y subunit alpha) [NCBI Gene 4800] {aka CBF-A, CBF-B, HAP2, NF-YA}, MPO (myeloperoxidase) [NCBI Gene 4353], SMARCD2 (SWI/SNF related BAF chromatin remodeling complex subunit D2) [NCBI Gene 6603] {aka BAF60B, CRACD2, PRO2451, Rsc6p, SGD2}, SMARCB1 (SWI/SNF related BAF chromatin remodeling complex subunit B1) [NCBI Gene 6598] {aka BAF47, CSS3, INI-1, INI1, MRD15, PPP1R144}, Myb (Myb proto-oncogene, transcription factor) [NCBI Gene 17863] {aka c-myb}, Cbfb (core binding factor beta) [NCBI Gene 12400] {aka PEA2, PEBP2b, Pebp2, Pebpb2}, Il3 (interleukin 3) [NCBI Gene 16187] {aka BPA, Csfmu, HCGF, Il-3, MCGF, PSF}, Mpo (myeloperoxidase) [NCBI Gene 17523] {aka mKIAA4033}, Nfyc (nuclear transcription factor-Y gamma) [NCBI Gene 18046], MIR130A (microRNA 130a) [NCBI Gene 406919] {aka MIRN130A, miRNA130A, mir-130a}
- **Diseases:** leukemias (MESH:D007938), infection (MESH:D007239), RAIN (MESH:C563663), cancers (MESH:D009369), MRE (MESH:D020238), acute myeloblastic leukemia (MESH:D015470)
- **Chemicals:** HCl (MESH:D006851), streptomycin (MESH:D013307), dithiothreitol (MESH:D004229), trypan blue (MESH:D014343), acetonitrile (MESH:C032159), lysine (MESH:D008239), penicillin (MESH:D010406), iodoacetamide (MESH:D007460), glycerol (MESH:D005990), L amino acids (MESH:D000596), formic acid (MESH:C030544), arginine (MESH:D001120), 13C6,15N2-lysine (-), trichloroacetic acid (MESH:D014238), hydrogen peroxide (MESH:D006861), proline (MESH:D011392), urea (MESH:D014508), acetone (MESH:D000096), oligonucleotide (MESH:D009841), glutamine (MESH:D005973), SDS (MESH:D012967), methanol (MESH:D000432)
- **Species:** Mus musculus (house mouse, species) [taxon 10090], Homo sapiens (human, species) [taxon 9606]
- **Mutations:** L for lysine, L for arginine, arginine-to-proline
- **Cell lines:** HeLa — Homo sapiens (Human), Human papillomavirus-related endocervical adenocarcinoma, Cancer cell line (CVCL_0030), 32Dcl3 — Mus musculus (Mouse), Factor-dependent cell line (CVCL_0119), S2 — Drosophila melanogaster (Fruit fly), Spontaneously immortalized cell line (CVCL_Z232), Kasumi-1 — Homo sapiens (Human), Childhood acute myeloid leukemia with maturation, Cancer cell line (CVCL_0589), CRL-11346 — Homo sapiens (Human), Finite cell line (CVCL_2U06)

## Full text

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## Figures

4 figures with captions in the complete paper: https://tomesphere.com/paper/PMC4659159/full.md

## References

56 references — full list in the complete paper: https://tomesphere.com/paper/PMC4659159/full.md

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Source: https://tomesphere.com/paper/PMC4659159