MicroRNA-216a is essential for cardiac angiogenesis

Rio P. Juni; Jordy M.M. Kocken; Ricardo C. Abreu; Lara Ottaviani; Tim Davalan; Burcu Duygu; Ella M. Poels; Aliaksei Vasilevich; Jana C. Hegenbarth; Mahesh Appari; Nicole Bitsch; Serve Olieslagers; Dorien M. Schrijvers; Monika Stoll; Joerg Heineke; Jan de Boer; Leon J. de Windt; Paula A. da Costa Martins

doi:10.1016/j.ymthe.2023.04.007

MicroRNA-216a is essential for cardiac angiogenesis

Rio P. Juni
, Jordy M.M. Kocken
, Ricardo C. Abreu
, Lara Ottaviani
, Tim Davalan
, Burcu Duygu
, Ella M. Poels
, Aliaksei Vasilevich
, Jana C. Hegenbarth
, Mahesh Appari
, Nicole Bitsch
, Serve Olieslagers
, Dorien M. Schrijvers
, Monika Stoll
, Joerg Heineke
, Jan de Boer
, Leon J. de Windt
, Paula A. da Costa Martins (Corresponding author)

Research output: Contribution to journal › Article › Academic › peer-review

11 Citations (Scopus)

58 Downloads (Pure)

Abstract

While it is experimentally supported that impaired myocardial vascularization contributes to a mismatch between myocardial oxygen demand and supply, a mechanistic basis for disruption of coordinated tissue growth and angiogenesis in heart failure remains poorly understood. Silencing strategies that impair microRNA biogenesis have firmly implicated microRNAs in the regulation of angiogenesis, and individual microRNAs prove to be crucial in developmental or tumor angiogenesis. A high-throughput functional screening for the analysis of a whole-genome microRNA silencing library with regard to their phenotypic effect on endothelial cell proliferation as a key parameter, revealed several anti- and pro-proliferative microRNAs. Among those was miR-216a, a pro-angiogenic microRNA which is enriched in cardiac microvascular endothelial cells and reduced in expression under cardiac stress conditions. miR-216a null mice display dramatic cardiac phenotypes related to impaired myocardial vascularization and unbalanced autophagy and inflammation, supporting a model where microRNA regulation of microvascularization impacts the cardiac response to stress.

Original language	English
Pages (from-to)	1807-1828
Number of pages	22
Journal	Molecular Therapy
Volume	31
Issue number	6
DOIs	https://doi.org/10.1016/j.ymthe.2023.04.007
Publication status	Published - 7 Jun 2023

Bibliographical note

Funding Information:
We acknowledge Genoway (Lyon, France) for helping with the strategy and generation of the miR-216a KO mice. Funding: A.V. acknowledges the financial support of the European Union's Seventh Framework (FP7/2007-2013) grant agreement 289720. J.D.B. acknowledges the financial support of the Dutch province of Limburg. L.D.W. acknowledges support from the Netherlands Cardiovascular Research Initiative and the Dutch Heart Foundation, the Netherlands. L.D.W. was further supported by grant 311549 from the European Research Council (ERC) and a VICI award 918-156-47 from The Netherlands Organization for Scientific Research (NWO). P.C.M. is supported by a MEERVOUD grant from The Netherlands Organization for Scientific Research (NWO), a Dutch CardioVascular Alliance (DCVA) awarded to the Phaedra consortium, as well as the Impulse Grant 2018 awarded to the Phaedra IMPACT consortium (CVON-2018-29), and is an Established Investigator of the Dutch Heart Foundation (NHS2015T066). P.C.M. and R.C.A. were further supported by the H2020 Twinning project RESETageing (GA 952266). J.M.M.K. was supported by a Dutch CardioVascular Alliance (DCVA) awarded to the Phaedra consortium as well as the Impulse Grant 2018 awarded to the Phaedra IMPACT consortium (CVON-2018-29). R.P.J. B.D. S.O. J.M.M.K. R.C.A. E.M.P. T.D. and P.A.d.C.M. performed northern blots, qRT-PCR, western blots experiments, and angiogenic functional assays. R.P.J. R.C.A. and A.V. performed the functional screening and respective validation. R.J.P. J.M.M.K. B.D. and M.A. performed primary cell isolations. R.J.P. D.M.S. and N.B. performed surgical procedures and echocardiography in mouse models. R.J.P. and J.M.M.K. performed histology in the different mouse models, the in situ hybridizations, and generated the luciferase reporter constructs used in the luciferase assays. R.J.P. T.D. and R.C.A. performed the photoacoustics combined with ultrasound analysis of the mouse models. L.O. J.C.H. and M.S. performed the sequencing and the bioinformatics analysis. P.A.d.C.M. and L.J.d.W. generated miR-216a gene-targeted mouse mice. J.D.B. and L.J.d.W. provided reagents, models or data, and critically reviewed the manuscript before submission. P.A.d.C.M. acquired funding for the study. L.d.W. and P.d.C.M. are cofounders of Mirabilis Therapeutics.

Funding

We acknowledge Genoway (Lyon, France) for helping with the strategy and generation of the miR-216a KO mice. Funding: A.V. acknowledges the financial support of the European Union's Seventh Framework (FP7/2007-2013) grant agreement 289720. J.D.B. acknowledges the financial support of the Dutch province of Limburg. L.D.W. acknowledges support from the Netherlands Cardiovascular Research Initiative and the Dutch Heart Foundation, the Netherlands. L.D.W. was further supported by grant 311549 from the European Research Council (ERC) and a VICI award 918-156-47 from The Netherlands Organization for Scientific Research (NWO). P.C.M. is supported by a MEERVOUD grant from The Netherlands Organization for Scientific Research (NWO), a Dutch CardioVascular Alliance (DCVA) awarded to the Phaedra consortium, as well as the Impulse Grant 2018 awarded to the Phaedra IMPACT consortium (CVON-2018-29), and is an Established Investigator of the Dutch Heart Foundation (NHS2015T066). P.C.M. and R.C.A. were further supported by the H2020 Twinning project RESETageing (GA 952266). J.M.M.K. was supported by a Dutch CardioVascular Alliance (DCVA) awarded to the Phaedra consortium as well as the Impulse Grant 2018 awarded to the Phaedra IMPACT consortium (CVON-2018-29). R.P.J. B.D. S.O. J.M.M.K. R.C.A. E.M.P. T.D. and P.A.d.C.M. performed northern blots, qRT-PCR, western blots experiments, and angiogenic functional assays. R.P.J. R.C.A. and A.V. performed the functional screening and respective validation. R.J.P. J.M.M.K. B.D. and M.A. performed primary cell isolations. R.J.P. D.M.S. and N.B. performed surgical procedures and echocardiography in mouse models. R.J.P. and J.M.M.K. performed histology in the different mouse models, the in situ hybridizations, and generated the luciferase reporter constructs used in the luciferase assays. R.J.P. T.D. and R.C.A. performed the photoacoustics combined with ultrasound analysis of the mouse models. L.O. J.C.H. and M.S. performed the sequencing and the bioinformatics analysis. P.A.d.C.M. and L.J.d.W. generated miR-216a gene-targeted mouse mice. J.D.B. and L.J.d.W. provided reagents, models or data, and critically reviewed the manuscript before submission. P.A.d.C.M. acquired funding for the study. L.d.W. and P.d.C.M. are cofounders of Mirabilis Therapeutics. We acknowledge Genoway (Lyon, France) for helping with the strategy and generation of the miR-216a KO mice. Funding: A.V. acknowledges the financial support of the European Union’s Seventh Framework ( FP7/2007-2013 ) grant agreement 289720 . J.D.B. acknowledges the financial support of the Dutch province of Limburg . L.D.W. acknowledges support from the Netherlands Cardiovascular Research Initiative and the Dutch Heart Foundation , the Netherlands. L.D.W. was further supported by grant 311549 from the European Research Council (ERC) and a VICI award 918-156-47 from The Netherlands Organization for Scientific Research ( NWO ). P.C.M. is supported by a MEERVOUD grant from The Netherlands Organization for Scientific Research ( NWO ), a Dutch CardioVascular Alliance ( DCVA ) awarded to the Phaedra consortium, as well as the Impulse Grant 2018 awarded to the Phaedra IMPACT consortium ( CVON-2018-29 ), and is an Established Investigator of the Dutch Heart Foundation ( NHS2015T066 ). P.C.M. and R.C.A. were further supported by the H2020 Twinning project RESETageing ( GA 952266 ). J.M.M.K. was supported by a Dutch CardioVascular Alliance ( DCVA ) awarded to the Phaedra consortium as well as the Impulse Grant 2018 awarded to the Phaedra IMPACT consortium ( CVON-2018-29 ).

Keywords

angiogenesis
autophagy
cardiac remodeling
endothelial cells
heart failure
microRNAs

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Cite this

Juni, R. P., Kocken, J. M. M., Abreu, R. C., Ottaviani, L., Davalan, T., Duygu, B., Poels, E. M., Vasilevich, A., Hegenbarth, J. C., Appari, M., Bitsch, N., Olieslagers, S., Schrijvers, D. M., Stoll, M., Heineke, J., de Boer, J., de Windt, L. J., & da Costa Martins, P. A. (2023). MicroRNA-216a is essential for cardiac angiogenesis. Molecular Therapy, 31(6), 1807-1828. https://doi.org/10.1016/j.ymthe.2023.04.007

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title = "MicroRNA-216a is essential for cardiac angiogenesis",

abstract = "While it is experimentally supported that impaired myocardial vascularization contributes to a mismatch between myocardial oxygen demand and supply, a mechanistic basis for disruption of coordinated tissue growth and angiogenesis in heart failure remains poorly understood. Silencing strategies that impair microRNA biogenesis have firmly implicated microRNAs in the regulation of angiogenesis, and individual microRNAs prove to be crucial in developmental or tumor angiogenesis. A high-throughput functional screening for the analysis of a whole-genome microRNA silencing library with regard to their phenotypic effect on endothelial cell proliferation as a key parameter, revealed several anti- and pro-proliferative microRNAs. Among those was miR-216a, a pro-angiogenic microRNA which is enriched in cardiac microvascular endothelial cells and reduced in expression under cardiac stress conditions. miR-216a null mice display dramatic cardiac phenotypes related to impaired myocardial vascularization and unbalanced autophagy and inflammation, supporting a model where microRNA regulation of microvascularization impacts the cardiac response to stress.",

keywords = "angiogenesis, autophagy, cardiac remodeling, endothelial cells, heart failure, microRNAs",

author = "Juni, \{Rio P.\} and Kocken, \{Jordy M.M.\} and Abreu, \{Ricardo C.\} and Lara Ottaviani and Tim Davalan and Burcu Duygu and Poels, \{Ella M.\} and Aliaksei Vasilevich and Hegenbarth, \{Jana C.\} and Mahesh Appari and Nicole Bitsch and Serve Olieslagers and Schrijvers, \{Dorien M.\} and Monika Stoll and Joerg Heineke and \{de Boer\}, Jan and \{de Windt\}, \{Leon J.\} and \{da Costa Martins\}, \{Paula A.\}",

note = "Funding Information: We acknowledge Genoway (Lyon, France) for helping with the strategy and generation of the miR-216a KO mice. Funding: A.V. acknowledges the financial support of the European Union's Seventh Framework (FP7/2007-2013) grant agreement 289720. J.D.B. acknowledges the financial support of the Dutch province of Limburg. L.D.W. acknowledges support from the Netherlands Cardiovascular Research Initiative and the Dutch Heart Foundation, the Netherlands. L.D.W. was further supported by grant 311549 from the European Research Council (ERC) and a VICI award 918-156-47 from The Netherlands Organization for Scientific Research (NWO). P.C.M. is supported by a MEERVOUD grant from The Netherlands Organization for Scientific Research (NWO), a Dutch CardioVascular Alliance (DCVA) awarded to the Phaedra consortium, as well as the Impulse Grant 2018 awarded to the Phaedra IMPACT consortium (CVON-2018-29), and is an Established Investigator of the Dutch Heart Foundation (NHS2015T066). P.C.M. and R.C.A. were further supported by the H2020 Twinning project RESETageing (GA 952266). J.M.M.K. was supported by a Dutch CardioVascular Alliance (DCVA) awarded to the Phaedra consortium as well as the Impulse Grant 2018 awarded to the Phaedra IMPACT consortium (CVON-2018-29). R.P.J. B.D. S.O. J.M.M.K. R.C.A. E.M.P. T.D. and P.A.d.C.M. performed northern blots, qRT-PCR, western blots experiments, and angiogenic functional assays. R.P.J. R.C.A. and A.V. performed the functional screening and respective validation. R.J.P. J.M.M.K. B.D. and M.A. performed primary cell isolations. R.J.P. D.M.S. and N.B. performed surgical procedures and echocardiography in mouse models. R.J.P. and J.M.M.K. performed histology in the different mouse models, the in situ hybridizations, and generated the luciferase reporter constructs used in the luciferase assays. R.J.P. T.D. and R.C.A. performed the photoacoustics combined with ultrasound analysis of the mouse models. L.O. J.C.H. and M.S. performed the sequencing and the bioinformatics analysis. P.A.d.C.M. and L.J.d.W. generated miR-216a gene-targeted mouse mice. J.D.B. and L.J.d.W. provided reagents, models or data, and critically reviewed the manuscript before submission. P.A.d.C.M. acquired funding for the study. L.d.W. and P.d.C.M. are cofounders of Mirabilis Therapeutics. ",

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doi = "10.1016/j.ymthe.2023.04.007",

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Juni, RP, Kocken, JMM, Abreu, RC, Ottaviani, L, Davalan, T, Duygu, B, Poels, EM, Vasilevich, A, Hegenbarth, JC, Appari, M, Bitsch, N, Olieslagers, S, Schrijvers, DM, Stoll, M, Heineke, J, de Boer, J, de Windt, LJ & da Costa Martins, PA 2023, 'MicroRNA-216a is essential for cardiac angiogenesis', Molecular Therapy, vol. 31, no. 6, pp. 1807-1828. https://doi.org/10.1016/j.ymthe.2023.04.007

TY - JOUR

T1 - MicroRNA-216a is essential for cardiac angiogenesis

AU - Juni, Rio P.

AU - Kocken, Jordy M.M.

AU - Abreu, Ricardo C.

AU - Ottaviani, Lara

AU - Davalan, Tim

AU - Duygu, Burcu

AU - Poels, Ella M.

AU - Vasilevich, Aliaksei

AU - Hegenbarth, Jana C.

AU - Appari, Mahesh

AU - Bitsch, Nicole

AU - Olieslagers, Serve

AU - Schrijvers, Dorien M.

AU - Stoll, Monika

AU - Heineke, Joerg

AU - de Boer, Jan

AU - de Windt, Leon J.

AU - da Costa Martins, Paula A.

N1 - Funding Information: We acknowledge Genoway (Lyon, France) for helping with the strategy and generation of the miR-216a KO mice. Funding: A.V. acknowledges the financial support of the European Union's Seventh Framework (FP7/2007-2013) grant agreement 289720. J.D.B. acknowledges the financial support of the Dutch province of Limburg. L.D.W. acknowledges support from the Netherlands Cardiovascular Research Initiative and the Dutch Heart Foundation, the Netherlands. L.D.W. was further supported by grant 311549 from the European Research Council (ERC) and a VICI award 918-156-47 from The Netherlands Organization for Scientific Research (NWO). P.C.M. is supported by a MEERVOUD grant from The Netherlands Organization for Scientific Research (NWO), a Dutch CardioVascular Alliance (DCVA) awarded to the Phaedra consortium, as well as the Impulse Grant 2018 awarded to the Phaedra IMPACT consortium (CVON-2018-29), and is an Established Investigator of the Dutch Heart Foundation (NHS2015T066). P.C.M. and R.C.A. were further supported by the H2020 Twinning project RESETageing (GA 952266). J.M.M.K. was supported by a Dutch CardioVascular Alliance (DCVA) awarded to the Phaedra consortium as well as the Impulse Grant 2018 awarded to the Phaedra IMPACT consortium (CVON-2018-29). R.P.J. B.D. S.O. J.M.M.K. R.C.A. E.M.P. T.D. and P.A.d.C.M. performed northern blots, qRT-PCR, western blots experiments, and angiogenic functional assays. R.P.J. R.C.A. and A.V. performed the functional screening and respective validation. R.J.P. J.M.M.K. B.D. and M.A. performed primary cell isolations. R.J.P. D.M.S. and N.B. performed surgical procedures and echocardiography in mouse models. R.J.P. and J.M.M.K. performed histology in the different mouse models, the in situ hybridizations, and generated the luciferase reporter constructs used in the luciferase assays. R.J.P. T.D. and R.C.A. performed the photoacoustics combined with ultrasound analysis of the mouse models. L.O. J.C.H. and M.S. performed the sequencing and the bioinformatics analysis. P.A.d.C.M. and L.J.d.W. generated miR-216a gene-targeted mouse mice. J.D.B. and L.J.d.W. provided reagents, models or data, and critically reviewed the manuscript before submission. P.A.d.C.M. acquired funding for the study. L.d.W. and P.d.C.M. are cofounders of Mirabilis Therapeutics.

PY - 2023/6/7

Y1 - 2023/6/7

N2 - While it is experimentally supported that impaired myocardial vascularization contributes to a mismatch between myocardial oxygen demand and supply, a mechanistic basis for disruption of coordinated tissue growth and angiogenesis in heart failure remains poorly understood. Silencing strategies that impair microRNA biogenesis have firmly implicated microRNAs in the regulation of angiogenesis, and individual microRNAs prove to be crucial in developmental or tumor angiogenesis. A high-throughput functional screening for the analysis of a whole-genome microRNA silencing library with regard to their phenotypic effect on endothelial cell proliferation as a key parameter, revealed several anti- and pro-proliferative microRNAs. Among those was miR-216a, a pro-angiogenic microRNA which is enriched in cardiac microvascular endothelial cells and reduced in expression under cardiac stress conditions. miR-216a null mice display dramatic cardiac phenotypes related to impaired myocardial vascularization and unbalanced autophagy and inflammation, supporting a model where microRNA regulation of microvascularization impacts the cardiac response to stress.

AB - While it is experimentally supported that impaired myocardial vascularization contributes to a mismatch between myocardial oxygen demand and supply, a mechanistic basis for disruption of coordinated tissue growth and angiogenesis in heart failure remains poorly understood. Silencing strategies that impair microRNA biogenesis have firmly implicated microRNAs in the regulation of angiogenesis, and individual microRNAs prove to be crucial in developmental or tumor angiogenesis. A high-throughput functional screening for the analysis of a whole-genome microRNA silencing library with regard to their phenotypic effect on endothelial cell proliferation as a key parameter, revealed several anti- and pro-proliferative microRNAs. Among those was miR-216a, a pro-angiogenic microRNA which is enriched in cardiac microvascular endothelial cells and reduced in expression under cardiac stress conditions. miR-216a null mice display dramatic cardiac phenotypes related to impaired myocardial vascularization and unbalanced autophagy and inflammation, supporting a model where microRNA regulation of microvascularization impacts the cardiac response to stress.

KW - angiogenesis

KW - autophagy

KW - cardiac remodeling

KW - endothelial cells

KW - heart failure

KW - microRNAs

UR - https://www.scopus.com/pages/publications/85153870231

U2 - 10.1016/j.ymthe.2023.04.007

DO - 10.1016/j.ymthe.2023.04.007

M3 - Article

C2 - 37073128

AN - SCOPUS:85153870231

SN - 1525-0016

VL - 31

SP - 1807

EP - 1828

JO - Molecular Therapy

JF - Molecular Therapy

IS - 6

ER -

MicroRNA-216a is essential for cardiac angiogenesis

Abstract

Bibliographical note

Funding

Keywords

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