miR-31 functions as a negative regulator of lymphatic vascular lineage-specific differentiation in vitro and vascular development in vivo

Institute of Pharmaceutical Sciences, Swiss Federal Institute of Technology, ETH Zürich, Wolfgang-Pauli-Str. 10, HCI H303, CH-8093 Zürich, Switzerland.
Molecular and Cellular Biology (Impact Factor: 5.04). 07/2010; 30(14):3620-34. DOI: 10.1128/MCB.00185-10
Source: PubMed

ABSTRACT The lymphatic vascular system maintains tissue fluid homeostasis, helps mediate afferent immune responses, and promotes cancer metastasis. To address the role microRNAs (miRNAs) play in the development and function of the lymphatic vascular system, we defined the in vitro miRNA expression profiles of primary human lymphatic endothelial cells (LECs) and blood vascular endothelial cells (BVECs) and identified four BVEC signature and two LEC signature miRNAs. Their vascular lineage-specific expression patterns were confirmed in vivo by quantitative real-time PCR and in situ hybridization. Functional characterization of the BVEC signature miRNA miR-31 identified a novel BVEC-specific posttranscriptional regulatory mechanism that inhibits the expression of lymphatic lineage-specific transcripts in vitro. We demonstrate that suppression of lymphatic differentiation is partially mediated via direct repression of PROX1, a transcription factor that functions as a master regulator of lymphatic lineage-specific differentiation. Finally, in vivo studies of Xenopus and zebrafish demonstrated that gain of miR-31 function impaired venous sprouting and lymphatic vascular development, thus highlighting the importance of miR-31 as a negative regulator of lymphatic development. Collectively, our findings identify miR-31 is a potent regulator of vascular lineage-specific differentiation and development in vertebrates.

Download full-text


Available from: André W Brändli, May 05, 2014
  • Source
    • "We previously established the Xenopus laevis tadpole as a genetic model for lymphangiogenesis research, phenocopying deficiencies of known mammalian lymphatic genes (Ny et al., 2005). We and others further applied the tadpole model to investigate molecular regulation of lymphatic vascular development, including its use in chemical library screens to identify anti-lymph/angiogenesis compounds (Kälin et al., 2009; Marino et al., 2011; Ny et al., 2008; Leslie Pedrioli et al., 2010). In these studies, visualization of the blood-and lymphatic vasculature depended on staining by in situ hybridization (ISH). "
    [Show abstract] [Hide abstract]
    ABSTRACT: The importance of the blood- and lymph vessels in the transport of essential fluids, gases, macromolecules and cells in vertebrates warrants optimal insight into the regulatory mechanisms underlying their development. Mouse and zebrafish models of lymphatic development are instrumental for gene discovery and gene characterization but are challenging for certain aspects, e.g. no direct accessibility of embryonic stages, or non-straightforward visualization of early lymphatic sprouting, respectively. We previously demonstrated that the Xenopus tadpole is a valuable model to study the processes of lymphatic development. However, a fluorescent Xenopus reporter directly visualizing the lymph vessels was lacking. Here, we created transgenic Tg(Flk1:eGFP) Xenopus laevis reporter lines expressing green fluorescent protein (GFP) in blood- and lymph vessels driven by the Flk1 (VEGFR-2) promoter. We also established a high-resolution fluorescent dye labeling technique selectively and persistently visualizing lymphatic endothelial cells, even in conditions of impaired lymph vessel formation or drainage function upon silencing of lymphangiogenic factors. Next, we applied the model to dynamically document blood and lymphatic sprouting and patterning of the initially avascular tadpole fin. Furthermore, quantifiable models of spontaneous or induced lymphatic sprouting into the tadpole fin were developed for dynamic analysis of loss-of-function and gain-of-function phenotypes using pharmacologic or genetic manipulation. Together with angiography and lymphangiography to assess functionality, Tg(Flk1:eGFP) reporter tadpoles readily allowed detailed lymphatic phenotyping of live tadpoles by fluorescence microscopy. The Tg(Flk1:eGFP) tadpoles represent a versatile model for functional lymph/angiogenomics and drug screening.
    Biology Open 09/2013; 2(9):882-90. DOI:10.1242/bio.20134739 · 2.42 Impact Factor
  • Source
    • "Loss of either claudin-like protein 24 (clp24) or synectin leads to defects in lymphatic development in both mice and zebrafish (Geudens et al., 2010; Saharinen et al., 2010). The microRNA miR-31 has been shown to be required for lymphatic gene expression, differentiation, and sprouting of human LECs and lymphatics in developing Xenopus and zebrafish (Pedrioli et al., 2010). The spleen tyrosine kinase (syk) and related zeta associated protein-70 (zap-70) are required for both angiogenic and lymphatigiogenic development in the fish (Christie et al., 2010). "
    [Show abstract] [Hide abstract]
    ABSTRACT: The lymphatic system is crucial for fluid homeostasis, immune responses, and numerous pathological processes. However, the molecular mechanisms responsible for establishing the anatomical form of the lymphatic vascular network remain largely unknown. Here, we show that chemokine signaling provides critical guidance cues directing early trunk lymphatic network assembly and patterning. The chemokine receptors Cxcr4a and Cxcr4b are expressed in lymphatic endothelium, whereas chemokine ligands Cxcl12a and Cxcl12b are expressed in adjacent tissues along which the developing lymphatics align. Loss- and gain-of-function studies in zebrafish demonstrate that chemokine signaling orchestrates the stepwise assembly of the trunk lymphatic network. In addition to providing evidence for a lymphatic vascular guidance mechanism, these results also suggest a molecular basis for the anatomical coalignment of lymphatic and blood vessels.
    Developmental Cell 04/2012; 22(4):824-36. DOI:10.1016/j.devcel.2012.01.011 · 10.37 Impact Factor
  • Source
    • "Whereas the human Prox1 open reading frame (ORF) is 2.2 kb long and encodes a 737-amino-acid-long protein (Oliver et al. 1993; Tomarev et al. 1998), the PROX1 gene is 50 kb long and expresses 8-kb-long transcripts in most organs and tissues, except retina (Tomarev et al. 1998) and testis (Steffensen et al. 2004), which express an 2.3-kb Prox1 mRNA. Detailed molecular analyses show that Prox1 mRNA harbors an 5.4-kb-long 3 0 UTR and that this evolutionarily conserved region of the gene is found to be subjected to posttranscriptional regulation by HuR (Yoo et al. 2010) and microRNAs (Kazenwadel et al. 2010; Pedrioli et al. 2010), indicating that Prox1 may be regulated by multiple physiological and pathological signals and stimuli. "
    [Show abstract] [Hide abstract]
    ABSTRACT: The blood and lymphatic systems are the two major circulatory systems in our body. Although the blood system has been studied extensively, the lymphatic system has received much less scientific and medical attention because of its elusive morphology and mysterious pathophysiology. However, a series of landmark discoveries made in the past decade has begun to change the previous misconception of the lymphatic system to be secondary to the more essential blood vascular system. In this article, we review the current understanding of the development and pathology of the lymphatic system. We hope to convince readers that the lymphatic system is no less essential than the blood circulatory system for human health and well-being.
    Cold Spring Harbor Perspectives in Medicine 04/2012; 2(4):a006445. DOI:10.1101/cshperspect.a006445 · 7.56 Impact Factor
Show more

Similar Publications