MicroRNA-126a Directs Lymphangiogenesis Through Interacting with Chemokine and Flt4 Signaling in Zebrafish

Jian Chen, Rong Fang Zhu, Fang Fang Li, Yu Lai Liang, Chen Wang, Yong Wen Qin, Shuang Huang, Xian Xian Zhao, Qing Jing

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

Objective - MicroRNA-126 (miR-126) is an endothelium-enriched miRNA and functions in vascular integrity and angiogenesis. The application of miRNA as potential biomarker and therapy target has been widely investigated in various pathological processes. However, its role in lymphatic diseases had not been widely explored. We aimed to reveal the role of miR-126 in lymphangiogenesis and the regulatory signaling pathways for potential targets of therapy. Approach and Results - Loss-of-function studies using morpholino oligonucleotides and CRISPR/Cas9 (clustered regularly interspaced short palindromic repeats/CRISPR-associated protein 9) system showed that silencing of miR-126a severely affected the formation of parachordal lymphangioblasts and thoracic duct in zebrafish embryos, although their development in miR-126b knockdown embryos was normal. Expression analyses by in situ hybridization and immunofluorescence indicated that miR-126a was expressed in lymphatic vessels, as well as in blood vessels. Time-lapse confocal imaging assay further revealed that knockdown of miR-126a blocked both lymphangiogenic sprouts budding from the posterior cardinal vein and lymphangioblasts extension along horizontal myoseptum. Bioinformatics analysis and in vivo report assay identified that miR-126a upregulated Cxcl12a by targeting its 5′ untranslated region. Moreover, loss- and gain-of-function studies revealed that Cxcl12a signaling acted downstream of miR-126a during parachordal lymphangioblast extension, whereby Flt4 signaling acts as a cooperator of miR-126a, allowing it to modulate lymphangiogenic sprout formation. Conclusions - These findings demonstrate that miR-126a directs lymphatic endothelial cell sprouting and extension by interacting with Cxcl12a-mediated chemokine signaling and Vegfc-Flt4 signal axis. Our results suggest that these key regulators of lymphangiogenesis may be involved in lymphatic pathogenesis of cardiovascular diseases.

Original languageEnglish (US)
Pages (from-to)2381-2393
Number of pages13
JournalArteriosclerosis, thrombosis, and vascular biology
Volume36
Issue number12
DOIs
StatePublished - Dec 1 2016

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Lymphangiogenesis
Zebrafish
MicroRNAs
Chemokines
CRISPR-Associated Proteins
Blood Vessels
Embryonic Structures
Clustered Regularly Interspaced Short Palindromic Repeats
Time-Lapse Imaging
Thoracic Duct
Morpholinos
Lymphatic Vessels
5' Untranslated Regions
Lymphatic Diseases
Pathologic Processes
Computational Biology
Endothelium
In Situ Hybridization
Fluorescent Antibody Technique
Veins

Keywords

  • endothelial cells
  • lymphangiogenesis
  • microRNAs
  • thoracic duct
  • zebrafish

ASJC Scopus subject areas

  • Cardiology and Cardiovascular Medicine

Cite this

MicroRNA-126a Directs Lymphangiogenesis Through Interacting with Chemokine and Flt4 Signaling in Zebrafish. / Chen, Jian; Zhu, Rong Fang; Li, Fang Fang; Liang, Yu Lai; Wang, Chen; Qin, Yong Wen; Huang, Shuang; Zhao, Xian Xian; Jing, Qing.

In: Arteriosclerosis, thrombosis, and vascular biology, Vol. 36, No. 12, 01.12.2016, p. 2381-2393.

Research output: Contribution to journalArticle

Chen, Jian ; Zhu, Rong Fang ; Li, Fang Fang ; Liang, Yu Lai ; Wang, Chen ; Qin, Yong Wen ; Huang, Shuang ; Zhao, Xian Xian ; Jing, Qing. / MicroRNA-126a Directs Lymphangiogenesis Through Interacting with Chemokine and Flt4 Signaling in Zebrafish. In: Arteriosclerosis, thrombosis, and vascular biology. 2016 ; Vol. 36, No. 12. pp. 2381-2393.
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abstract = "Objective - MicroRNA-126 (miR-126) is an endothelium-enriched miRNA and functions in vascular integrity and angiogenesis. The application of miRNA as potential biomarker and therapy target has been widely investigated in various pathological processes. However, its role in lymphatic diseases had not been widely explored. We aimed to reveal the role of miR-126 in lymphangiogenesis and the regulatory signaling pathways for potential targets of therapy. Approach and Results - Loss-of-function studies using morpholino oligonucleotides and CRISPR/Cas9 (clustered regularly interspaced short palindromic repeats/CRISPR-associated protein 9) system showed that silencing of miR-126a severely affected the formation of parachordal lymphangioblasts and thoracic duct in zebrafish embryos, although their development in miR-126b knockdown embryos was normal. Expression analyses by in situ hybridization and immunofluorescence indicated that miR-126a was expressed in lymphatic vessels, as well as in blood vessels. Time-lapse confocal imaging assay further revealed that knockdown of miR-126a blocked both lymphangiogenic sprouts budding from the posterior cardinal vein and lymphangioblasts extension along horizontal myoseptum. Bioinformatics analysis and in vivo report assay identified that miR-126a upregulated Cxcl12a by targeting its 5′ untranslated region. Moreover, loss- and gain-of-function studies revealed that Cxcl12a signaling acted downstream of miR-126a during parachordal lymphangioblast extension, whereby Flt4 signaling acts as a cooperator of miR-126a, allowing it to modulate lymphangiogenic sprout formation. Conclusions - These findings demonstrate that miR-126a directs lymphatic endothelial cell sprouting and extension by interacting with Cxcl12a-mediated chemokine signaling and Vegfc-Flt4 signal axis. Our results suggest that these key regulators of lymphangiogenesis may be involved in lymphatic pathogenesis of cardiovascular diseases.",
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AU - Chen, Jian

AU - Zhu, Rong Fang

AU - Li, Fang Fang

AU - Liang, Yu Lai

AU - Wang, Chen

AU - Qin, Yong Wen

AU - Huang, Shuang

AU - Zhao, Xian Xian

AU - Jing, Qing

PY - 2016/12/1

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N2 - Objective - MicroRNA-126 (miR-126) is an endothelium-enriched miRNA and functions in vascular integrity and angiogenesis. The application of miRNA as potential biomarker and therapy target has been widely investigated in various pathological processes. However, its role in lymphatic diseases had not been widely explored. We aimed to reveal the role of miR-126 in lymphangiogenesis and the regulatory signaling pathways for potential targets of therapy. Approach and Results - Loss-of-function studies using morpholino oligonucleotides and CRISPR/Cas9 (clustered regularly interspaced short palindromic repeats/CRISPR-associated protein 9) system showed that silencing of miR-126a severely affected the formation of parachordal lymphangioblasts and thoracic duct in zebrafish embryos, although their development in miR-126b knockdown embryos was normal. Expression analyses by in situ hybridization and immunofluorescence indicated that miR-126a was expressed in lymphatic vessels, as well as in blood vessels. Time-lapse confocal imaging assay further revealed that knockdown of miR-126a blocked both lymphangiogenic sprouts budding from the posterior cardinal vein and lymphangioblasts extension along horizontal myoseptum. Bioinformatics analysis and in vivo report assay identified that miR-126a upregulated Cxcl12a by targeting its 5′ untranslated region. Moreover, loss- and gain-of-function studies revealed that Cxcl12a signaling acted downstream of miR-126a during parachordal lymphangioblast extension, whereby Flt4 signaling acts as a cooperator of miR-126a, allowing it to modulate lymphangiogenic sprout formation. Conclusions - These findings demonstrate that miR-126a directs lymphatic endothelial cell sprouting and extension by interacting with Cxcl12a-mediated chemokine signaling and Vegfc-Flt4 signal axis. Our results suggest that these key regulators of lymphangiogenesis may be involved in lymphatic pathogenesis of cardiovascular diseases.

AB - Objective - MicroRNA-126 (miR-126) is an endothelium-enriched miRNA and functions in vascular integrity and angiogenesis. The application of miRNA as potential biomarker and therapy target has been widely investigated in various pathological processes. However, its role in lymphatic diseases had not been widely explored. We aimed to reveal the role of miR-126 in lymphangiogenesis and the regulatory signaling pathways for potential targets of therapy. Approach and Results - Loss-of-function studies using morpholino oligonucleotides and CRISPR/Cas9 (clustered regularly interspaced short palindromic repeats/CRISPR-associated protein 9) system showed that silencing of miR-126a severely affected the formation of parachordal lymphangioblasts and thoracic duct in zebrafish embryos, although their development in miR-126b knockdown embryos was normal. Expression analyses by in situ hybridization and immunofluorescence indicated that miR-126a was expressed in lymphatic vessels, as well as in blood vessels. Time-lapse confocal imaging assay further revealed that knockdown of miR-126a blocked both lymphangiogenic sprouts budding from the posterior cardinal vein and lymphangioblasts extension along horizontal myoseptum. Bioinformatics analysis and in vivo report assay identified that miR-126a upregulated Cxcl12a by targeting its 5′ untranslated region. Moreover, loss- and gain-of-function studies revealed that Cxcl12a signaling acted downstream of miR-126a during parachordal lymphangioblast extension, whereby Flt4 signaling acts as a cooperator of miR-126a, allowing it to modulate lymphangiogenic sprout formation. Conclusions - These findings demonstrate that miR-126a directs lymphatic endothelial cell sprouting and extension by interacting with Cxcl12a-mediated chemokine signaling and Vegfc-Flt4 signal axis. Our results suggest that these key regulators of lymphangiogenesis may be involved in lymphatic pathogenesis of cardiovascular diseases.

KW - endothelial cells

KW - lymphangiogenesis

KW - microRNAs

KW - thoracic duct

KW - zebrafish

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