Roles of non-coding RNA in megakaryocytopoiesis and thrombopoiesis: new target therapies in ITP

Figures & data

Figure 1. miRNAs regulate the differentiation of hematopoietic stem cells to MKs to produce platelets. miRNAs act on the 3‘end of mRNAs to inhibit their degradation or translation of proteins by targeting the MRE.

Abbreviations: MRE: microRNA response element; HSC: hematopoietic stem cells; MKs: megakaryocytes; PL: platelets.

Table I. Functions and mechanisms of long non-coding RNAs.

Long non-coding RNA	Function	Interacting with DNA, miRNA, proteins	Physiopathological process
UMLILO [51]	Regulation of transcription	WDR5–MLL	Activates transcription of CXCL chemokines.
APOLO [52]		DNA (R-loops); LHP1	Functions as a decoy for LHP1 at the R-loop; regulates expression of auxin-responsive genes.
CHASERR [53]		CHD2	Represses Chd2 expression in cis and broadly regulates CHD2 targets.
SWINGN [54]		SWI/SNF	Activates the proximal gene GAS6 and distant genes in trans through chromatin looping.
lncPRESS1 [55]		Sirtuin 6	Functions as a decoy for sirtuin 6 in regulating gene expression.
LincRNA-p21 [56]		P21	lincRNA-p21 acts in concert with hnRNP-K as a coactivator for p53-dependent p21 transcription, thereby the p53 tumor suppressor pathway.
REG1CP [57]		REG3A	REG1CP forms an RNA-DNA triad with REG3A to recruit FANCJ helicase for REG3A transcription, and promotes tumorigenesis.
HOTAIR [58]	Post-transcriptional regulation	miR-613	Sequesters miR-613, thereby causing upregulation of notch3 mRNAs and promoting the development of Prostate cancer.
PVT1 [59]		miR-26b	Sponging miR-26b promotes Melanoma Progression.
MT1P3 [20]		miR-126	Sponging miR-126 promotes p2y12 expression and activate platelet.
FAST [60]		β-TrCP	Interacts with β-TrCP to prevent β-catenin degradation, thereby activating WNT signaling and promoting pluripotency by maintaining the WNT pathway in human ESCs.
PYCARD-AS1 [61]		PYCARD	PYCARD-AS1 interact with the PYCARD mRNA each other via their 5’ overlapping region, leading to inhibition of PYCARD translation.
NORAD [62]	Genome integrity	Pumilio	Maintains genomic stability by sequestering PUMILIO proteins, which repress the stability and translation of mRNAs to which they bind.
DINO [63]		p53	Bound p53 protein and promoted its stabilization, mediating a p53 auto-amplification loop.
CONCR [64]		DDX11	Interacts with DDX11 to modulate cell cycle progression and DNA replication.
FIRRE [65]	Structural functions	hnRNPU	Interacts with multiple chromosomal regions through hnRNPU and involved in gene regulation in trans, regulates hematopoiesis in vivo.
sno-lncRNAs [66]		RBFOX2	Associates with RBFOX2 and regulates mRNA splicing.

Figure 2. LncAS-RBM15 participation in the regulation of megakaryocytopoiesis and thrombopoiesis. LncAS-RBM15 combines with RBM15-mRNA and enhances RBM15 protein translation. The transcription factor RUNX1 can activate the expression of both RBM15 and AS-RBM15.

Abbreviations: RUNX1:RUNX family transcription factor 1; HSC: Hematopoietic Stem Cell, MKs: Megakaryocytes, PL: Platelets.

Table II. Examples of mechanistically studied miRNAs and lncRNAs with roles during hematopoiesis.

Name	Function	Mechanism	Reference
MiR-130a	Megakaryocyte differentiation	Regulates the transcription factor MAFB to promote megakaryocyte differentiation	[37]
MiR-10a	Megakaryocyte differentiation	Regulates the transcription factor HOXA1 to promote megakaryocyte differentiation	[37]
MiR-223-3p	Megakaryocyte polyploidy	Regulation of human myosin heavy chain 10 (MYH10) to promote megakaryocyte polyploidy	[39]
MiR-146a	Megakaryocyte polyploidy	Targeting CXCR4 impedes its translation Inhibits megakaryocyte polyploidy formation	[40,41]
MiR-125b	Megakaryocyte maturation and differentiation	Regulates the cell cycle and promotes megakaryocyte maturation and differentiation	[42,43]
MiR-22	Megakaryocyte differentiation	Regulates the transcription of growth factor independent 1 (GFI1) and promotes megakaryocyte differentiation	[44]
MiR-34a	Megakaryocyte colony	Promotes hematopoietic stem cells (HSPC) to form megakaryocyte colonies to stimulate megakaryopoiesis	[45,46]
MiR-150	Megakaryocyte differentiation	Regulation of transcription factor MYB promotes megakaryocyte differentiation	[47]
MiR-155	Megakaryocyte differentiation	Regulates the cell cycle and inhibits the differentiation of hematopoietic stem cells into megakaryocytes	[48,49]
MiR-125a-5p	Megakaryocyte migration and proplatelet production	Actin-binding protein L-plastin, encoded by LCP1, inhibits megakaryocyte migration and proplatelet formation by reducing its expression level	[50]
LncRNAAS-RBM15	Megakaryocyte differentiation	Regulates RBM15 protein synthesis and inhibits megakaryocyte differentiation	[94–96]
LncRNA NEAT1	Megakaryocyte differentiation and platelet production	Regulates the cell cycle, promotes megakaryocyte differentiation and platelet granule formation	[97]

Table III. Examples of mechanistically studied miRNAs and lncRNAs with roles during ITP.

Name	Function	Mechanism	Reference
MiR-55	Participate in ITP development	Targeted regulatory cytokine 1 (SOCS1) promotes ITP	[121]
MiR-130a		Targeting transforming growth factor (TGF-β1) and interleukin 18 (IL-18) promotes ITP	[122]
MiR-155-5p		Upregulation of SOCS1 promotes PD1/PDL1 pathway mediated macrophage M2 polarization to prevent ITP	[123]
MiR-106b-5p		Treg/Th17 immune imbalance in immune cells induced by ITP via NR4A3/Foxp3 pathway	[124]
MiR-557		Inhibit megakaryocyte differentiation and maturation	[125]
LncRNA TMEVPG1		Stimulates IFN-γ transcription to slow down the progression of ITP	[127]
LncRNA MEG3		Inhibits miRNA-125a-5p expression and induces immune imbalance of immune cell Treg/Th17 in ITP	[128]
LncRNA GAS5		Promoting STAT3 ubiquitination inhibits immune cell Th17 differentiation and alleviates ITP	[129]
LncRNA PVT1		Targeting NOTCH1 makes immune cell Treg/Th17 imbalance to promote the occurrence of ITP	[130]

Fanucchi S, Fok ET, Dalla E, Shibayama Y, Borner K, Chang EY, Stoychev S, Imakaev M, Grimm D, Wang KC, et al. Immune genes are primed for robust transcription by proximal long noncoding RNAs located in nuclear compartments. Nat Genet. 2019;51(1):138–150. doi:10.1038/s41588-018-0298-2. Epub 2018/12/12.

 PubMed Web of Science ®Google Scholar

Ariel F, Lucero L, Christ A, Mammarella MF, Jegu T, Veluchamy A, Mariappan K, Latrasse D, Blein T, Liu C, et al. R-loop mediated trans action of the APOLO long noncoding RNA. Mol Cell. 2020;77(5):1055–1065 e1054. doi:10.1016/j.molcel.2019.12.015. Epub 2020/01/19.

 PubMed Web of Science ®Google Scholar

Rom A, Melamed L, Gil N, Goldrich MJ, Kadir R, Golan M, Biton I, Perry RBT, Ulitsky I. Regulation of CHD2 expression by the Chaserr long noncoding RNA gene is essential for viability. Nat Commun. 2019;10(1). doi:10.1038/s41467-019-13075-8.

 PubMedGoogle Scholar

Grossi E, Raimondi I, Goni E, Gonzalez J, Marchese FP, Chapaprieta V, Martin-Subero JI, Guo S, Huarte M. A lncRNA-SWI/SNF complex crosstalk controls transcriptional activation at specific promoter regions. Nat Commun. 2020;11(1):936. doi:10.1038/s41467-020-14623-3. Epub 2020/02/20.

 PubMed Web of Science ®Google Scholar

Jain AK, Xi YX, McCarthy R, Allton K, Akdemir KC, Patel LR, Aronow B, Lin CR, Li W, Yang LQ, et al. LncPRESS1 is a p53-regulated LncRNA that safeguards pluripotency by disrupting SIRT6-mediated De-acetylation of Histone H3K56. Mol Cell. 2016;64(5):967–981. doi:10.1016/j.molcel.2016.10.039.

 PubMed Web of Science ®Google Scholar

Dimitrova N, Zamudio JR, Jong RM, Soukup D, Resnick R, Sarma K, Ward AJ, Raj A, Lee JT, Sharp PA, et al. LincRNA-p21 activates p21 in cis to promote Polycomb target gene expression and to enforce the G1/S checkpoint. Mol Cell. 2014;54(5):777–790. doi:10.1016/j.molcel.2014.04.025. Epub 2014/05/27.

 PubMed Web of Science ®Google Scholar

Yari H, Jin L, Teng L, Wang YF, Wu YY, Liu GZ, Gao W, Liang J, Xi YF, Feng YC, et al. LncRNA REG1CP promotes tumorigenesis through an enhancer complex to recruit FANCJ helicase for REG3A transcription. Nat Commun. 2019;10(1). doi:10.1038/s41467-019-13313-z.

 PubMedGoogle Scholar

Cai H, Yao J, An Y, Chen X, Chen W, Wu D, Luo B, Yang Y, Jiang Y, Sun D, et al. LncRNA HOTAIR acts a competing endogenous RNA to control the expression of notch3 via sponging miR-613 in pancreatic cancer. Oncotarget. 2017;8(20):32905–32917. doi:10.18632/oncotarget.16462. Epub 2017/04/19.

 PubMedGoogle Scholar

Wang BJ, Ding HW, Ma GA. Long noncoding RNA PVT1 promotes melanoma progression via endogenous sponging miR-26b. Oncol Res. 2018;26(5):675–681. doi:10.3727/096504017X14920318811730. Epub 2017/04/15.

 PubMed Web of Science ®Google Scholar

Zhou M, Gao M, Luo Y, Gui R, Ji H. Long non-coding RNA metallothionein 1 pseudogene 3 promotes p2y12 expression by sponging miR-126 to activate platelet in diabetic animal model. Platelets. 2019;30(4):452–459. doi:10.1080/09537104.2018.1457781.

 PubMed Web of Science ®Google Scholar

Guo CJ, Ma XK, Xing YH, Zheng CC, Xu YF, Shan L, Zhang J, Wang S, Wang Y, Carmichael GG, et al. Distinct processing of lncRNAs contributes to non-conserved functions in stem cells. Cell. 2020;181(3):621–636 e622. doi:10.1016/j.cell.2020.03.006. Epub 2020/04/08.

 PubMed Web of Science ®Google Scholar

Miao H, Wang LL, Zhan HM, Dai JS, Chang YB, Wu F, Liu T, Liu ZY, Gao CF, Li L, et al. A long noncoding RNA distributed in both nucleus and cytoplasm operates in the PYCARD-regulated apoptosis by coordinating the epigenetic and translational regulation. PLoS Genet. 2019;15(5):e1008144. doi:10.1371/journal.pgen.1008144.

 PubMed Web of Science ®Google Scholar

Lee S, Kopp F, Chang TC, Sataluri A, Chen B, Sivakumar S, Yu H, Xie Y, Mendell JT. Noncoding RNA NORAD regulates genomic stability by sequestering PUMILIO proteins. Cell. 2016;164(1–2):69–80. doi:10.1016/j.cell.2015.12.017.

 PubMed Web of Science ®Google Scholar

Schmitt AM, Garcia JT, Hung T, Flynn RA, Shen Y, Qu K, Payumo AY, Peres-da-Silva A, Broz DK, Baum R, et al. An inducible long noncoding RNA amplifies DNA damage signaling. Nat Genet. 2016;48(11):1370–1376. doi:10.1038/ng.3673. Epub 2016/10/28.

 PubMed Web of Science ®Google Scholar

Marchese FP, Grossi E, Marin-Bejar O, Bharti SK, Raimondi I, Gonzalez J, Martinez-Herrera DJ, Athie A, Amadoz A, Brosh RM, Jr., et al. A long noncoding RNA regulates sister chromatid cohesion. Mol Cell. 2016;63(3):397–407. doi:10.1016/j.molcel.2016.06.031. Epub 2016/08/02.

 PubMed Web of Science ®Google Scholar

Lewandowski JP, Lee JC, Hwang T, Sunwoo H, Goldstein JM, Groff AF, Chang NP, Mallard W, Williams A, Henao-Meija J, et al. The Firre locus produces a trans-acting RNA molecule that functions in hematopoiesis. Nat Commun. 2019;10(1). doi:10.1038/s41467-019-12970-4.

 Google Scholar

Yin QF, Yang L, Zhang Y, Xiang JF, Wu YW, Carmichael GG, Chen LL. Long noncoding RNAs with snoRNA ends. Mol Cell. 2012;48(2):219–230. doi:10.1016/j.molcel.2012.07.033. Epub 2012/09/11.

 PubMed Web of Science ®Google Scholar

Garzon R, Pichiorri F, Palumbo T, Iuliano R, Cimmino A, Aqeilan R, Volinia S, Bhatt D, Alder H, Marcucci G, et al. MicroRNA fingerprints during human megakaryocytopoiesis. Proc Natl Acad Sci USA. 2006;103(13):5078–5083. doi:10.1073/pnas.0600587103.

 PubMed Web of Science ®Google Scholar

Shi R, Zhou X, Ji WJ, Zhang YY, Ma YQ, Zhang JQ, Li YM. The emerging role of miR-223 in platelet reactivity: implications in antiplatelet therapy. Biomed Res Int. 2015;2015:981841. doi:10.1155/2015/981841. Epub 2015/07/30.

 PubMed Web of Science ®Google Scholar

Opalinska JB, Bersenev A, Zhang Z, Schmaier AA, Choi J, Yao Y, D’Souza J, Tong W, Weiss MJ. MicroRNA expression in maturing murine megakaryocytes. Blood. 2010;116(23):e128–138. doi:10.1182/blood-2010-06-292920. Epub 2010/08/20.

 PubMed Web of Science ®Google Scholar

Labbaye C, Spinello I, Quaranta MT, Pelosi E, Pasquini L, Petrucci E, Biffoni M, Nuzzolo ER, Billi M, Foa R, et al. A three-step pathway comprising PLZF/miR-146a/cxcr4 controls megakaryopoiesis. Nat Cell Biol. 2008;10(7):788–801. doi:10.1038/ncb1741. Epub 2008/06/24.

 PubMed Web of Science ®Google Scholar

Kandi R, Undi R, Gutti RK. MiR-125b regulates cell proliferation and survival in neonatal megakaryocytes. Ann Hematol. 2014;93(6):1065–1066. doi:10.1007/s00277-013-1928-5. Epub 2013/10/26.

 PubMed Web of Science ®Google Scholar

Qu M, Fang F, Zou X, Zeng Q, Fan Z, Chen L, Yue W, Xie X, Pei X. miR-125b modulates megakaryocyte maturation by targeting the cell-cycle inhibitor p19(ink4d). Cell Death Dis. 2016;7(10):e2430. doi:10.1038/cddis.2016.288. Epub 2016/10/21.

 PubMedGoogle Scholar

Weiss CN, Ito K. microRNA-22 promotes megakaryocyte differentiation through repression of its target, GFI1. Blood Adv. 2019;3(1):33–46. doi:10.1182/bloodadvances.2018023804. Epub 2019/01/09.

 PubMed Web of Science ®Google Scholar

Ichimura A, Ruike Y, Terasawa K, Shimizu K, Tsujimoto G. MicroRNA-34a inhibits cell proliferation by repressing mitogen-activated protein kinase kinase 1 during megakaryocytic differentiation of K562 cells. Mol Pharmacol. 2010;77(6):1016–1024. doi:10.1124/mol.109.063321. Epub 2010/03/20.

 PubMed Web of Science ®Google Scholar

Navarro F, Gutman D, Meire E, Caceres M, Rigoutsos I, Bentwich Z, Lieberman J. miR-34a contributes to megakaryocytic differentiation of K562 cells independently of p53. Blood. 2009;114(10):2181–2192. doi:10.1182/blood-2009-02-205062. Epub 2009/07/09.

 PubMed Web of Science ®Google Scholar

Emambokus N, Vegiopoulos A, Harman B, Jenkinson E, Anderson G, Frampton J. Progression through key stages of haemopoiesis is dependent on distinct threshold levels of c-Myb. EMBO J. 2003;22:4478–4488. doi:10.1093/emboj/cdg434. Epub 2003/08/28.

 PubMed Web of Science ®Google Scholar

Lu J, Guo S, Ebert BL, Zhang H, Peng X, Bosco J, Pretz J, Schlanger R, Wang JY, Mak RH, et al. MicroRNA-mediated control of cell fate in megakaryocyte-erythrocyte progenitors. Dev Cell. 2008;14(6):843–853. doi:10.1016/j.devcel.2008.03.012. Epub 2008/06/10.

 PubMed Web of Science ®Google Scholar

Georgantas RW, 3rd, Hildreth R, Morisot S, Alder J, Liu CG, Heimfeld S, Calin GA, Croce CM, Civin CI. CD34+ hematopoietic stem-progenitor cell microRNA expression and function: a circuit diagram of differentiation control. Proc Natl Acad Sci USA. 2007;104(8):2750–2755. doi:10.1073/pnas.0610983104. Epub 2007/02/13.

 PubMed Web of Science ®Google Scholar

Bhatlekar S, Manne BK, Basak I, Edelstein LC, Tugolukova E, Stoller ML, Cody MJ, Morley SC, Nagalla S, Weyrich AS, et al. miR-125a-5p regulates megakaryocyte proplatelet formation via the actin-bundling protein L-plastin. Blood. 2020;136(15):1760–1772. doi:10.1182/blood.2020005230. Epub 2020/08/28.

 PubMed Web of Science ®Google Scholar

Bian W, Chen W, Jiang X, Qu H, Jiang J, Yang J, Liang X, Zhao B, Sun Y, Zhang C. Downregulation of long non-coding RNA nuclear Paraspeckle assembly transcript 1 inhibits MEG-01 differentiation and platelet-like particles activity. Front Genet. 2020;11:571467. doi:10.3389/fgene.2020.571467. Epub 2020/11/17.

 PubMed Web of Science ®Google Scholar

Qian BH, Ye X, Zhang L, Sun Y, Zhang JR, Gu ML, Qin Q, Chen J, Deng AM. Increased miR-155 expression in peripheral blood mononuclear cells of primary immune thrombocytopenia patients was correlated with serum cytokine profiles. Acta Haematol. 2015;133(3):257–263. doi:10.1159/000362150. Epub 2014/11/22.

 PubMed Web of Science ®Google Scholar

Zhao H, Li H, Du W, Zhang D, Ge J, Xue F, Zhou Z, Yang R. Reduced MIR130A is involved in primary immune thrombocytopenia via targeting TGFB1 and IL18. Br J Haematol. 2014;166(5):767–773. doi:10.1111/bjh.12934. Epub 2014/05/08.

 PubMed Web of Science ®Google Scholar

Chang Y, Chen X, Tian Y, Gao X, Liu Z, Dong X, Wang L, He F, Zhou J. Downregulation of microRNA-155-5p prevents immune thrombocytopenia by promoting macrophage M2 polarization via the SOCS1-dependent PD1/PDL1 pathway. Life Sci. 2020;257:118057. doi:10.1016/j.lfs.2020.118057. Epub 2020/07/08.

 PubMed Web of Science ®Google Scholar

Li JQ, Tian JM, Fan XR, Wang ZY, Ling J, Wu XF, Yang FY, Xia YL. miR-106b-5p induces immune imbalance of Treg/Th17 in immune thrombocytopenic purpura through NR4A3/Foxp3 pathway. Cell Cycle. 2020;19(11):1265–1274. doi:10.1080/15384101.2020.1746485. Epub 2020/04/24.

 PubMed Web of Science ®Google Scholar

Wang Y, Guo Y, Zhang X, Zhao H, Zhang B, Wu Y, Zhang J. The role and mechanism of miR-557 in inhibiting the differentiation and maturation of megakaryocytes in immune thrombocytopenia. RNA Biol. 2021;18(11):1953–1968. doi:10.1080/15476286.2021.1884783. Epub 2021/02/16.

 PubMed Web of Science ®Google Scholar

Li H, Hao Y, Zhang D, Fu R, Liu W, Zhang X, Xue F, Yang R. Aberrant expression of long noncoding RNA TMEVPG1 in patients with primary immune thrombocytopenia. Autoimmunity. 2016;49(7):496–502. doi:10.3109/08916934.2016.1167192. Epub 2016/04/07.

 PubMed Web of Science ®Google Scholar

Li JQ, Hu SY, Wang ZY, Lin J, Jian S, Dong YC, Wu XF, Dai L, Cao LJ. Long non-coding RNA MEG3 inhibits microRNA-125a-5p expression and induces immune imbalance of Treg/Th17 in immune thrombocytopenic purpura. Biomed Pharmacother. 2016;83:905–911. doi:10.1016/j.biopha.2016.07.057. Epub 2016/10/25.

 PubMed Web of Science ®Google Scholar

Li J, Tian J, Lu J, Wang Z, Ling J, Wu X, Yang F, Xia Y. LncRNA GAS5 inhibits Th17 differentiation and alleviates immune thrombocytopenia via promoting the ubiquitination of STAT3. Int Immunopharmacol. 2020;80:106127. doi:10.1016/j.intimp.2019.106127. Epub 2020/01/25.

 PubMed Web of Science ®Google Scholar

Huang Y, Qiao Y, Zhao Y, Li Y, Yuan J, Zhou J, Sun H, Wang H. Large scale RNA-binding proteins/LncRNAs interaction analysis to uncover lncRNA nuclear localization mechanisms. Brief Bioinform. 2021;22(6): bbab195. doi:10.1093/bib/bbab195.

 PubMed Web of Science ®Google Scholar