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

Abstract

Noncoding RNAs (ncRNAs) are a group of RNA molecules that cannot encode proteins, and a better understanding of the complex interaction networks coordinated by ncRNAs will provide a theoretical basis for the development of therapeutics targeting the regulatory effects of ncRNAs. Platelets are produced upon the differentiation of hematopoietic stem cells into megakaryocytes, 10¹¹ per day, and are renewed every 8–9 days. The process of thrombopoiesis is affected by multiple factors, in which ncRNAs also exert a significant regulatory role. This article reviewed the regulatory roles of ncRNAs, mainly microRNAs (miRNAs), circRNAs (circular RNAs), and long non-coding RNAs (lncRNAs), in thrombopoiesis in recent years as well as their roles in primary immune thrombocytopenia (ITP).

Plain Language Summary

What is the context?

• Platelets are produced from progenitor cells named megakaryocytes (MKs) differentiated from bone marrow-derived hematopoietic stem cells (HSCs).

• Thrombopoiesis refers to the process by which platelet-producing MKs release platelet granules into peripheral blood under the shear force of blood flow for further development and maturation.

• The process of megakaryocytopoiesis and thrombopoiesis is affected by multiple factors, wherein some ncRNAs also exert a significant regulatory role.

• miRNAs/lncRNAs play a promising role in t primary immune thrombocytopenia (ITP).

What is new?

• This article reviewed the regulatory roles of ncRNAs, mainly microRNAs (miRNAs), circRNAs (circular RNAs), and long non-coding RNAs (lncRNAs), in thrombopoiesis.

• This article also reviewed the roles of ncRNAs in ITP.

What is the impact?

Changes in ncRNA expression are associated with changes in the production of MKs, thrombopoiesis, and platelet function, which allows a new understanding of the pathogenesis of many congenital or acquired platelet-related diseases.

Keywords:

• Megakaryocytopoiesis
• non-coding RNA, thrombopoiesis

Introduction

Noncoding RNAs (ncRNAs), a class of RNAs that do not encode proteins [1], are actually transcribed by most of the mammalian genome [2], and act as key regulators of physiological processes in the human body [3,4]. Therefore, an in-depth understanding of the complicated interaction network coordinated by non-coding RNAs will provide a substantial theoretical basis for developing therapeutic measures targeting the regulatory role of ncRNAs [5,6]. Platelets are produced from progenitor cells named megakaryocytes (MKs) differentiated from bone marrow-derived hematopoietic stem cells (HSCs) under the action of a series of hematopoietic growth factors [7,8]. At a steady state, MKs can support a daily production of 10¹¹ platelets with a renewal every 8–9 days. This process is greatly influenced by environmental alterations, which may lead to a more than 10-fold increase in the production responding to stimulation [9]. Thrombopoiesis refers to the process by which platelet-producing MKs release platelet granules into peripheral blood under the shear force of blood flow for further development and maturation [10–12]. Thrombocytopenia is a disorder characterized by defective or low platelet production with a risk of possible life-threatening bleeding. Although a variety of key factors have been identified to mediate MK maturation and platelet release [13–16], the regulatory mechanism of thrombopoiesis is still undefined. Despite lacking a nucleus, platelets possess multiple types of RNA inherited from MKs, encompassing protein-coding messenger RNAs (mRNAs) and ncRNAs such as microRNAs (miRNAs) and long noncoding RNAs (lncRNAs) [17,18], which have the major features of post-transcriptional RNA metabolism [19,20]. The process of thrombopoiesis is affected by multiple factors, wherein some ncRNAs also exert a significant regulatory role. Herein, this study systematically reviewed the research progress on the regulatory roles of non-coding RNAs, mainly miRNAs, and lncRNAs, on thrombopoiesis in recent years.

miRNA-mediated regulation of thrombopoiesis

MiRNAs are a category of endogenous small RNAs that are approximately 20–24 nt in length [21]. The majority of miRNAs are localized in intronic regions that are highly conserved in different species [22,23] and are highly homologous in sequence, which is strongly linked to the significance of its function [24], exhibiting fine-tuned control of protein expression through recognition, binding, and translational repression of complementary sequences to protein-coding mRNA transcripts [25,26]. The diverse functions and expression patterns of miRNAs in platelets or MKs reveal that they may play key modulatory roles in the biology of platelets. Circulating platelets are essential in the physiology of blood coagulation that maintains the hemostatic balance and protects vascular integrity [27]. Conversely, abnormality in platelet reactivity is related to diverse pathological factors such as cardiovascular disorders, inflammation, etc., leading to morbidity and mortality [28–31]. Multiple miRNAs have been uncovered in platelets, but their roles remain greatly unknown. One such miRNA, miR-223, is documented to be the most abundant miRNA in platelets [32,33], followed by miR-126 [20,34]. MiR-96, miR-200b, miR-495, miR-107, and miR-223 are underscored to closely correlate with reactivity, aggregation, secretion, and adhesion of platelets [35].

In recent years, miRNAs have gradually attracted attention, and changes in the miRNA expression signatures in cancers have been observed, rendering them prognostic and auxiliary diagnostic predictors [36]. Additionally, miRNAs are important mediators of hematopoietic progenitor cell differentiation into MKs in the hematopoietic system [37]. Present research focusing on the regulatory mechanism underlying the MK differentiation is the most in-depth stage across the whole process of thrombopoiesis, in which the action of miRNAs in the differentiation of MKs has also been gradually excavated. For instance, miR-125b and miR-660 can augment the number of polyploid MKs, but reversely, overexpression of the miR-23a/27A/24-2 cluster in human HSCs attenuates the MK maturation- and thrombopoiesis-related clone formation [38]; miR-223-3p stimulates MK polyploidy via mediating the expression of human myosin heavy chain 10 (MYH10) [39]; however, miR-146a inhibits the polyploidy in MKs [40,41]; miR-125b expedites the maturation and differentiation of MKs by controlling cell cycle [42,43]; miR-130a and miR-10a facilitate MK differentiation by regulating transcription factors MAFB and HOXA1, respectively, and both the two miRNAs are up-regulated in acute megakaryocytic leukemia cell lines [37]; miR-22 inhibits the transcription of growth factor independent-1 (GFI1) to accelerate MK differentiation [44]; miR-34a stimulates megakaryopoiesis by promoting the formation of MK colonies from HSCs and its expression is also increased during induction of MK differentiation in K562 cells [45,46]; miR-150 facilitates MK differentiation by targeting the transcription factor MYB [47]; reversely, miR-155 restrains the differentiation of HSCs into MKs through cell cycle regulation [48,49]; miR-125a-5p acts on LCP1-encoded actin-binding protein L-plastin to reduce its expression, thereby affecting MK migration and proplatelet formation [50] (Table I). Since the discovery of miRNAs in platelets, regulatory roles of more and more miRNAs have been successively reported, revealing new regulatory mechanisms during MK differentiation (Figure 1). Increasing attention has been paid to the expression patterns of different platelet miRNAs in a healthy or diseased status, and despite unique challenges, there is plenty of room for improvement in the research field of miRNA-dependent regulation of MK-platelet to shed light on the unknown molecular mechanisms that potentially modulate thrombopoiesis and platelet function.

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.

LncRNA-mediated regulation of thrombopoiesis

Long non-coding RNAs (lncRNAs) refer to a class of RNAs longer than 200 nt that are not translated into proteins [67]. LncRNAs exert regulatory effects on the expression of genes by binding to DNA, RNA, and proteins [68–70], which are also referred to as competing endogenous RNAs (ceRNAs) such as HOTAIR [58], PTV1 [59], HOXa11-AS [71], PFL [72], and ARSR [73]. LncRNA-miRNA interaction networks seem to be a major theme that existed in the biology of lncRNAs, which have been proven to be either the source or suppressors of miRNAs. LncRNAs play a significant role in the fine-tuning translation machinery and its regulation by modulating the key functions of other ncRNAs such as miRNAs [74]. LncRNAs are derived not only from their own promoters but also from promoters shared with differently transcribed coding or non-coding genes, or from enhancer sequences [75,76]. LncRNAs have a unique evolutionary conserved pattern with higher tissue specificity [77]. For years, studies have unraveled that multiple regulatory paradigms controlled by several established lncRNAs affect a wide range of cellular activities and are functionally relevant to normal development and pathophysiology in multiple diseases [78,79], including several types of cancers, neurological and cardiovascular diseases, immune and metabolic disorders. Several types of lncRNAs have been identified based on different parameters such as transcript length, association with annotated protein-coding genes,

and similarity to mRNA [80]. These classes of RNA molecules are distinct from other RNAs relying on some unique characteristics, including alternative forms of biogenesis [81], unique regulatory mechanism [82], cis-regulatory activity, and RNA-binding domain with functional structure [83,84] (Table I). Although lncRNAs have attracted attention from different research teams, much is still unknown about their functions and structures.

LncRNAs play a crucial role in maintaining the human HSC population and are well-known to be critical for the identification of specific differentiation pathways of HSCs. For example, lncHSC-2 regulates the self-renewal and differentiation of HSCs [85]. Another lncRNA shlnc-EC6 is related to erythroid differentiation [86]. HOTAIRM1 acts as a cell cycle regulator during myeloid maturation in addition to granulocyte differentiation [87,88]. EGO positively regulates several genes essential for eosinophil production [89]. LINC00173 can stimulate granulocyte differentiation and myeloid colony formation [90]. Hematopoietic development shows similar processes in both mice and humans [91]. However, the lncRNAs involved in mouse and human hematopoiesis are poorly conserved, suggesting different evolution patterns of lncRNAs [92]. Nevertheless, the implications of lncRNAs in MK differentiation and thrombopoiesis represent a largely unexplored area. Megakaryopoiesis is controlled by intricate networks involving bone marrow hematopoietic growth factors [93]. Among diverse regulators, lncRNAs are also a group of critical regulators for the induction of MKs and thrombopoiesis (Table II). Little is known about the roles of lncRNAs in megakaryopoiesis. Many past studies have revealed that some lncRNAs are abnormally expressed in myeloid progenitors and MKs. LncRNA AS-RBM15, which is situated on chromosome 1 with a total length of 52 797 bp, transcribed in the antisense orientation, and overlapped with the 5’ untranslated region (UTR) of the protein-coding gene RNA-binding protein 15 (RBM15), is initially unveiled to control the differentiation of MKs [98]. RBM15 is highly expressed during hematopoiesis, particularly in myeloid differentiation and other hematopoietic cell lines, in which its mechanism is mediated by RBP-Jkappa-dependent Notch signaling activation, but lowly expressed in MKs [94]. Differential expression of RBM15 crucially functions in the development of myeloid, MKs, and progenitor cell subsets as well as dysfunction-related disorders [95]. There is also evidence in another study supporting the involvement of this antisense RNA transcript lncRNA AS-RBM15 in the transcription and translation regulation of the RBM15 protein in MKs [96]. Activation of lncRNA AS-RBM15 plays a direct inducing role in the protein synthesis of RBM15, an extremely important control component of MK-terminal differentiation [99] (Figure 2). Besides, the transcription factor RUNX1 restrains the erythroid gene expression program during MK differentiation [100] and maintains the activities of RBM15 and AS-RBM15 as a consequence of erythroid gene expression patterns. These genes are restricted by epigenetic repression of the master erythroid regulator, krueppel-like factor 1 (KLF1), during MK differentiation [101]. Furthermore, the current study shows the high AS-RBM15 expression reflecting that the function of RUNX1 affects the balance between erythroid and MK differentiation [80]. In addition, a recent study has reported that lncRNA NEAT1 participates in MK differentiation and thrombopoiesis and its mechanism of action affects the cell cycle and promotes the differentiation of MEG-01 cells and production of platelet granules [97]. There is scientific evidence demonstrating that the biological functions of lncRNAs cannot be ignored and integrated complementary approaches are demanded for these critical lncRNAs. These approaches are varied depending on their modes of action in the development of MKs and disease processes. Furthermore, these lncRNAs may be implicated in multiple pathways related to the developmental process of MKs.

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]

Abundant circular RNAs expression in differentiating megakaryocytes

CircRNAs are a subgroup of highly stable non-coding RNA, which are dominantly generated from pre-mRNA back-splicing [102]. Recently, studies have shown several potential functions of circRNAs, which can interact with RNA-binding proteins or modulate transcription by sponging miRNAs [103,104]. Furthermore, circRNAs are currently recognized as important regulatory molecules in cell differentiation, cell proliferation, and apoptosis [105]. However, the functions of circRNAs during hematopoietic differentiation are still not clear. High-throughput sequencing revealed that circRNAs are highly enriched in human platelets [106]. The mRNAs decay more rapidly than circRNAs in platelet formation. Therefore, circRNAs are hugely enriched in platelets compared with megakaryocytes. By comparing the expression pattern of circRNAs and mRNAs, circRNAs were demonstrated to modulate hematopoietic differentiation and their expression significantly increased in differentiating megakaryocytes [107]. Xia et al. identify a circular RNA named cia-cGAS, which can regulate the balance between self-renewal and differentiation of hematopoietic stem cells [108]. These results suggest that circRNAs might play important role in megakaryocytopoiesis and thrombopoiesis.

The roles of ncRNAs in primary immune thrombocytopenia

Primary immune thrombocytopenia (ITP) is a clinically common, acquired autoimmune hemorrhagic disease in the absence of obvious exogenous factors, predominantly characterized by increased platelet destruction, peripheral blood thrombocytopenia, and normal MKs or accompanied by maturation disorder. This disease frequently recurs and persists and can be divided into acute and chronic disorders according to the duration of the disease: acute ITP patients are more commonly present in children often with a history of infection before the onset, showing a relatively short disease course, most of which can be self-cured; chronic ITP is most often diagnosed in adults with unclear etiology, slow onset, and frequent recurrence that is difficult to relieve spontaneously [109,110]. As an autoimmune disease, abnormal immune responses develop in vivo following ITP, yet its pathogenesis is still unclear [111]. It is currently believed that the pathogenesis of ITP is a multi-factorial process. In addition to the involvement of cellular immunity, humoral immunity, MK maturation disorder, and the complement system, some recently identified new mechanisms may be suggested to participate in the onset of ITP, such as the dysregulation of miRNAs/lncRNAs [112], FcγR-independent platelet clearance pathway [113,114], soluble programmed cell death receptor-1 (sPD-1) [115–117], and metalloproteinase 10 (ADAM10) [118,119]. Existing research data have elucidated an immunoregulatory effect of miRNAs/lncRNAs on immune response and immune cells, indicating that miRNAs/lncRNAs play a promising role in the pathogenesis of ITP. The regulatory role of miRNAs/lncRNAs in thrombopoiesis has recently become a research hotspot. Currently, the effect of miRNAs/lncRNAs on platelet function in ITP patients is still undetermined. Accumulating studies have suggested that abnormally expressed miRNAs/lncRNAs are involved in the pathogenesis of ITP [120]. One such miRNA, miR-55, targets the suppressor of cytokine signaling 1 (SOCS1), promoting the occurrence of ITP [121]; miR-130a is also involved in the development of ITP via targeting transforming growth factor (TGF-β1) and interleukin 18 (IL-18) [122]; silencing miR-155-5p upregulates SOCS1 and accelerates PD1/PDL1 pathway-mediated M2 macrophage polarization to prevent the development of ITP [123]; miR-106b-5p induces the imbalance of immune cells Treg/Th17 in ITP through the NR4A3/Foxp3 pathway [124]; miR-557 suppresses the differentiation and maturation of MKs in ITP [125]; miR-409-3p expression is upregulated in the PBMCs of ITP patients after effective treatment, and IFN-γ is confirmed to be a target gene of miR-409-3p [126]. LncRNA TMEVPG1 stimulates IFN-γ transcription to slow down the progression of ITP [127]. LncRNA MEG3 inhibits the miR-125a-5p expression and triggers an immune imbalance of Treg/Th17 in ITP [128]; LncRNA GAS5 alleviates ITP by motivating STAT3 ubiquitination and inhibiting the differentiation of immune cells Th17 [129]; LncRNA PVT1 targets NOTCH1 to elicit the immune imbalance of Treg/Th17 ratio, facilitating the occurrence of ITP [130]. With a deepened understanding of ncRNAs, it is still worth exploring the significance of ncRNAs in the pathogenesis of ITP; for instance, whether the differential expression of ncRNAs in plasma can serve as specific indicators of ITP (Table III). All these issues require an illustration through more thorough fundamental research and clinical observations. Therapeutic targeting of miRNAs and lncRNAs represents a promising approach for the treatment of many diseases such as cardiovascular disease and cancer. Over the past few decades, several RNA-based therapies, including ASO anti-microRNAs (antimiRs), short hairpin RNAs (shRNAs), small interfering RNAs (siRNAs), and antisense oligonucleotides (ASOs) have been trying to translate into clinical application. However, only more than ten RNA-based therapeutics are approved by the Food and Drug Administration (FDA) and the European Medicines Agency (EMA) until now. All these RNA drugs are either ASOs or siRNAs, which can cause specific gene degradation in the central nervous system, muscle, or liver [131]. Currently, no therapeutics targeting miRNAs and lncRNAs in megakaryopoiesis and thrombopoiesis enter the clinic. In the future, the issues associated with tolerability, delivery methods, and specificity must be overcome.

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]

Perspectives

With the changing understanding of ncRNA based on the continuous deepening exploration, ncRNAs have gradually emerged as important regulators of gene expression in the human body; the aberrant expression of ncRNA has become a pathogenic factor to the human body and also they serve as targets for the treatment of various human diseases. At present, much is unknown about the multiple functions, corresponding targets, and specific molecular mechanisms of ncRNAs. Although thousands of ncRNAs are expressed during hematopoiesis, studies on the functions of specific ncRNAs with experimental validation are limited. The increasing demand to decipher the exact functions of ncRNAs in the development of MKs and thrombopoiesis raises further research questions. Up to now, the significance of ncRNAs in megakaryocytopoiesis in relation to lineage-specific development has only been described in a few studies. Under different physiological conditions, an in-depth understanding of the changes in the ncRNA transcriptome in MKs, particularly how these changes indicate their importance in the development and progression of MKs, may contribute to the identification of new cellular pathways related to the generation of mature MKs from hematopoietic stem/progenitor cells (HSPCs). Understanding the ncRNA profile is also of crucial importance to provide information to directly reflect MK lineage relationship and functional similarities to the hematopoietic system. Furthermore, deep excavation of the ncRNA profiles in the MK population is also an efficient way to identify and predict their novel developmental roles in terminal megakaryocytopoiesis and subsequent platelet production.

Taken together, changes in ncRNA expression are associated with changes in the production of MKs, thrombopoiesis, and platelet function. In recent years, scientists have developed methods to generate platelets by in vitro inducing the differentiation of umbilical cord blood stem cells, bone marrow HSCs, human induced pluripotent stem cells, and mesenchymal stem cells into MKs. Nevertheless, due to limited stem cell sources, deficient platelet production, and other factors such as the potential carcinogenic risk of induced pluripotent stem cells, these studies are still in the laboratory exploration stage, unable to achieve mass production for clinical application. Research on MK differentiation suggests the possibility of producing clinical-grade platelets in vitro on a large scale. Meanwhile, the improvement of platelet isolation technology aids in improving the quality and quantity of platelets. Great progress has been made in the investigation of molecular mechanisms, which allows a new understanding of the pathogenesis of many congenital or acquired platelet-related diseases.

Acknowledgments

We thank the Home for Researchers editorial team (www.home-for-researchers.com) for editing the language of the manuscript.

Disclosure statement

No potential conflict of interest was reported by the authors.

Additional information

Funding

This work was supported by Provincial Natural Science Foundation of Shandong Province, China [Grant No. ZR2021MH082].