A systems approach delivers a functional microRNA catalog and expanded targets for seizure suppression in temporal lobe epilepsy

Morten T. Venø; Cristina R. Reschke; Gareth Morris; Niamh M.C. Connolly; Junyi Su; Yan Yan; Tobias Engel; Eva M. Jimenez-Mateos; Lea M. Harder; Dennis Pultz; Stefan J. Haunsberger; Ajay Pal; Janosch P. Heller; Aoife Campbell; Elena Langa; Gary P. Brennan; Karen Conboy; Amy Richardson; Braxton A. Norwood; Lara S. Costard; Valentin Neubert; Federico Del Gallo; Beatrice Salvetti; Vamshidhar R. Vangoor; Amaya Sanz-Rodriguez; Juha Muilu; Paolo F. Fabene; R. Jeroen Pasterkamp; Jochen H.M. Prehn; Stephanie Schorge; Jens S. Andersen; Felix Rosenow; Sebastian Bauer; Jørgen Kjems; David C. Henshall

doi:10.1073/pnas.1919313117

A systems approach delivers a functional microRNA catalog and expanded targets for seizure suppression in temporal lobe epilepsy

Morten T. Venø, Cristina R. Reschke, Gareth Morris, Niamh M.C. Connolly, Junyi Su, Yan Yan, Tobias Engel, Eva M. Jimenez-Mateos, Lea M. Harder, Dennis Pultz, Stefan J. Haunsberger, Ajay Pal, Janosch P. Heller, Aoife Campbell, Elena Langa, Gary P. Brennan, Karen Conboy, Amy Richardson, Braxton A. Norwood, Lara S. CostardValentin Neubert, Federico Del Gallo, Beatrice Salvetti, Vamshidhar R. Vangoor, Amaya Sanz-Rodriguez, Juha Muilu, Paolo F. Fabene, R. Jeroen Pasterkamp, Jochen H.M. Prehn, Stephanie Schorge, Jens S. Andersen, Felix Rosenow, Sebastian Bauer, Jørgen Kjems, David C. Henshall^*

^*Corresponding author for this work

Research output: Contribution to journal/Conference contribution in journal/Contribution to newspaper › Journal article › Research › peer-review

43 Citations (Scopus)

71 Downloads (Pure)

Abstract

Temporal lobe epilepsy is the most common drug-resistant form of epilepsy in adults. The reorganization of neural networks and the gene expression landscape underlying pathophysiologic network behavior in brain structures such as the hippocampus has been suggested to be controlled, in part, by microRNAs. To systematically assess their significance, we sequenced Argonaute-loaded microRNAs to define functionally engaged microRNAs in the hippocampus of three different animal models in two species and at six time points between the initial precipitating insult through to the establishment of chronic epilepsy. We then selected commonly up-regulated microRNAs for a functional in vivo therapeutic screen using oligonucleotide inhibitors. Argonaute sequencing generated 1.44 billion small RNA reads of which up to 82% were microRNAs, with over 400 unique microRNAs detected per model. Approximately half of the detected microRNAs were dysregulated in each epilepsy model. We prioritized commonly up-regulated microRNAs that were fully conserved in humans and designed custom antisense oligonucleotides for these candidate targets. Antiseizure phenotypes were observed upon knockdown of miR-10a-5p, miR-21a-5p, and miR-142a- 5p and electrophysiological analyses indicated broad safety of this approach. Combined inhibition of these three microRNAs reduced spontaneous seizures in epileptic mice. Proteomic data, RNA sequencing, and pathway analysis on predicted and validated targets of these microRNAs implicated derepressed TGF-β signaling as a shared seizure-modifying mechanism. Correspondingly, inhibition of TGF- β signaling occluded the antiseizure effects of the antagomirs. Together, these results identify shared, dysregulated, and functionally active microRNAs during the pathogenesis of epilepsy which represent therapeutic antiseizure targets.

Original language	English
Journal	Proceedings of the National Academy of Sciences (PNAS)
Volume	117
Issue	27
Pages (from-to)	15977-15988
Number of pages	12
ISSN	0027-8424
DOIs	https://doi.org/10.1073/pnas.1919313117
Publication status	Published - Jul 2020

Keywords

Antisense oligonucleotide
Biomarker
Epigenetic
Epilepsy
Noncoding RNA

Access to Document

10.1073/pnas.1919313117Licence: CC BY-NC-ND

15977.fullFinal published version, 2.98 MBLicence: CC BY-NC-ND

Cite this

Venø, M. T., Reschke, C. R., Morris, G., Connolly, N. M. C., Su, J., Yan, Y., Engel, T., Jimenez-Mateos, E. M., Harder, L. M., Pultz, D., Haunsberger, S. J., Pal, A., Heller, J. P., Campbell, A., Langa, E., Brennan, G. P., Conboy, K., Richardson, A., Norwood, B. A., ... Henshall, D. C. (2020). A systems approach delivers a functional microRNA catalog and expanded targets for seizure suppression in temporal lobe epilepsy. Proceedings of the National Academy of Sciences (PNAS), 117(27), 15977-15988. https://doi.org/10.1073/pnas.1919313117

@article{d7d173c4c327400a8a792f736821df0f,

title = "A systems approach delivers a functional microRNA catalog and expanded targets for seizure suppression in temporal lobe epilepsy",

abstract = "Temporal lobe epilepsy is the most common drug-resistant form of epilepsy in adults. The reorganization of neural networks and the gene expression landscape underlying pathophysiologic network behavior in brain structures such as the hippocampus has been suggested to be controlled, in part, by microRNAs. To systematically assess their significance, we sequenced Argonaute-loaded microRNAs to define functionally engaged microRNAs in the hippocampus of three different animal models in two species and at six time points between the initial precipitating insult through to the establishment of chronic epilepsy. We then selected commonly up-regulated microRNAs for a functional in vivo therapeutic screen using oligonucleotide inhibitors. Argonaute sequencing generated 1.44 billion small RNA reads of which up to 82% were microRNAs, with over 400 unique microRNAs detected per model. Approximately half of the detected microRNAs were dysregulated in each epilepsy model. We prioritized commonly up-regulated microRNAs that were fully conserved in humans and designed custom antisense oligonucleotides for these candidate targets. Antiseizure phenotypes were observed upon knockdown of miR-10a-5p, miR-21a-5p, and miR-142a- 5p and electrophysiological analyses indicated broad safety of this approach. Combined inhibition of these three microRNAs reduced spontaneous seizures in epileptic mice. Proteomic data, RNA sequencing, and pathway analysis on predicted and validated targets of these microRNAs implicated derepressed TGF-β signaling as a shared seizure-modifying mechanism. Correspondingly, inhibition of TGF- β signaling occluded the antiseizure effects of the antagomirs. Together, these results identify shared, dysregulated, and functionally active microRNAs during the pathogenesis of epilepsy which represent therapeutic antiseizure targets.",

keywords = "Antisense oligonucleotide, Biomarker, Epigenetic, Epilepsy, Noncoding RNA",

author = "Ven{\o}, {Morten T.} and Reschke, {Cristina R.} and Gareth Morris and Connolly, {Niamh M.C.} and Junyi Su and Yan Yan and Tobias Engel and Jimenez-Mateos, {Eva M.} and Harder, {Lea M.} and Dennis Pultz and Haunsberger, {Stefan J.} and Ajay Pal and Heller, {Janosch P.} and Aoife Campbell and Elena Langa and Brennan, {Gary P.} and Karen Conboy and Amy Richardson and Norwood, {Braxton A.} and Costard, {Lara S.} and Valentin Neubert and Gallo, {Federico Del} and Beatrice Salvetti and Vangoor, {Vamshidhar R.} and Amaya Sanz-Rodriguez and Juha Muilu and Fabene, {Paolo F.} and Pasterkamp, {R. Jeroen} and Prehn, {Jochen H.M.} and Stephanie Schorge and Andersen, {Jens S.} and Felix Rosenow and Sebastian Bauer and J{\o}rgen Kjems and Henshall, {David C.}",

year = "2020",

month = jul,

doi = "10.1073/pnas.1919313117",

language = "English",

volume = "117",

pages = "15977--15988",

journal = "Proceedings of the National Academy of Sciences (PNAS)",

issn = "0027-8424",

publisher = "National Academy of Sciences",

number = "27",

}

Venø, MT, Reschke, CR, Morris, G, Connolly, NMC, Su, J, Yan, Y, Engel, T, Jimenez-Mateos, EM, Harder, LM, Pultz, D, Haunsberger, SJ, Pal, A, Heller, JP, Campbell, A, Langa, E, Brennan, GP, Conboy, K, Richardson, A, Norwood, BA, Costard, LS, Neubert, V, Gallo, FD, Salvetti, B, Vangoor, VR, Sanz-Rodriguez, A, Muilu, J, Fabene, PF, Pasterkamp, RJ, Prehn, JHM, Schorge, S, Andersen, JS, Rosenow, F, Bauer, S, Kjems, J & Henshall, DC 2020, 'A systems approach delivers a functional microRNA catalog and expanded targets for seizure suppression in temporal lobe epilepsy', Proceedings of the National Academy of Sciences (PNAS), vol. 117, no. 27, pp. 15977-15988. https://doi.org/10.1073/pnas.1919313117

A systems approach delivers a functional microRNA catalog and expanded targets for seizure suppression in temporal lobe epilepsy. / Venø, Morten T.; Reschke, Cristina R.; Morris, Gareth et al.
In: Proceedings of the National Academy of Sciences (PNAS), Vol. 117, No. 27, 07.2020, p. 15977-15988.

Research output: Contribution to journal/Conference contribution in journal/Contribution to newspaper › Journal article › Research › peer-review

TY - JOUR

T1 - A systems approach delivers a functional microRNA catalog and expanded targets for seizure suppression in temporal lobe epilepsy

AU - Venø, Morten T.

AU - Reschke, Cristina R.

AU - Morris, Gareth

AU - Connolly, Niamh M.C.

AU - Su, Junyi

AU - Yan, Yan

AU - Engel, Tobias

AU - Jimenez-Mateos, Eva M.

AU - Harder, Lea M.

AU - Pultz, Dennis

AU - Haunsberger, Stefan J.

AU - Pal, Ajay

AU - Heller, Janosch P.

AU - Campbell, Aoife

AU - Langa, Elena

AU - Brennan, Gary P.

AU - Conboy, Karen

AU - Richardson, Amy

AU - Norwood, Braxton A.

AU - Costard, Lara S.

AU - Neubert, Valentin

AU - Gallo, Federico Del

AU - Salvetti, Beatrice

AU - Vangoor, Vamshidhar R.

AU - Sanz-Rodriguez, Amaya

AU - Muilu, Juha

AU - Fabene, Paolo F.

AU - Pasterkamp, R. Jeroen

AU - Prehn, Jochen H.M.

AU - Schorge, Stephanie

AU - Andersen, Jens S.

AU - Rosenow, Felix

AU - Bauer, Sebastian

AU - Kjems, Jørgen

AU - Henshall, David C.

PY - 2020/7

Y1 - 2020/7

N2 - Temporal lobe epilepsy is the most common drug-resistant form of epilepsy in adults. The reorganization of neural networks and the gene expression landscape underlying pathophysiologic network behavior in brain structures such as the hippocampus has been suggested to be controlled, in part, by microRNAs. To systematically assess their significance, we sequenced Argonaute-loaded microRNAs to define functionally engaged microRNAs in the hippocampus of three different animal models in two species and at six time points between the initial precipitating insult through to the establishment of chronic epilepsy. We then selected commonly up-regulated microRNAs for a functional in vivo therapeutic screen using oligonucleotide inhibitors. Argonaute sequencing generated 1.44 billion small RNA reads of which up to 82% were microRNAs, with over 400 unique microRNAs detected per model. Approximately half of the detected microRNAs were dysregulated in each epilepsy model. We prioritized commonly up-regulated microRNAs that were fully conserved in humans and designed custom antisense oligonucleotides for these candidate targets. Antiseizure phenotypes were observed upon knockdown of miR-10a-5p, miR-21a-5p, and miR-142a- 5p and electrophysiological analyses indicated broad safety of this approach. Combined inhibition of these three microRNAs reduced spontaneous seizures in epileptic mice. Proteomic data, RNA sequencing, and pathway analysis on predicted and validated targets of these microRNAs implicated derepressed TGF-β signaling as a shared seizure-modifying mechanism. Correspondingly, inhibition of TGF- β signaling occluded the antiseizure effects of the antagomirs. Together, these results identify shared, dysregulated, and functionally active microRNAs during the pathogenesis of epilepsy which represent therapeutic antiseizure targets.

AB - Temporal lobe epilepsy is the most common drug-resistant form of epilepsy in adults. The reorganization of neural networks and the gene expression landscape underlying pathophysiologic network behavior in brain structures such as the hippocampus has been suggested to be controlled, in part, by microRNAs. To systematically assess their significance, we sequenced Argonaute-loaded microRNAs to define functionally engaged microRNAs in the hippocampus of three different animal models in two species and at six time points between the initial precipitating insult through to the establishment of chronic epilepsy. We then selected commonly up-regulated microRNAs for a functional in vivo therapeutic screen using oligonucleotide inhibitors. Argonaute sequencing generated 1.44 billion small RNA reads of which up to 82% were microRNAs, with over 400 unique microRNAs detected per model. Approximately half of the detected microRNAs were dysregulated in each epilepsy model. We prioritized commonly up-regulated microRNAs that were fully conserved in humans and designed custom antisense oligonucleotides for these candidate targets. Antiseizure phenotypes were observed upon knockdown of miR-10a-5p, miR-21a-5p, and miR-142a- 5p and electrophysiological analyses indicated broad safety of this approach. Combined inhibition of these three microRNAs reduced spontaneous seizures in epileptic mice. Proteomic data, RNA sequencing, and pathway analysis on predicted and validated targets of these microRNAs implicated derepressed TGF-β signaling as a shared seizure-modifying mechanism. Correspondingly, inhibition of TGF- β signaling occluded the antiseizure effects of the antagomirs. Together, these results identify shared, dysregulated, and functionally active microRNAs during the pathogenesis of epilepsy which represent therapeutic antiseizure targets.

KW - Antisense oligonucleotide

KW - Biomarker

KW - Epigenetic

KW - Epilepsy

KW - Noncoding RNA

UR - http://www.scopus.com/inward/record.url?scp=85088209858&partnerID=8YFLogxK

U2 - 10.1073/pnas.1919313117

DO - 10.1073/pnas.1919313117

M3 - Journal article

C2 - 32581127

AN - SCOPUS:85088209858

SN - 0027-8424

VL - 117

SP - 15977

EP - 15988

JO - Proceedings of the National Academy of Sciences (PNAS)

JF - Proceedings of the National Academy of Sciences (PNAS)

IS - 27

ER -

A systems approach delivers a functional microRNA catalog and expanded targets for seizure suppression in temporal lobe epilepsy

Abstract

Keywords

Access to Document

Other files and links

Fingerprint

Cite this