Adenosine kinase as a target for therapeutic antisense strategies in epilepsy

Panos Theofilas, Sukhmani Brar, Kerry-Ann Mitchell, Hai Ying Shen, Ursula S. Sandau, David Poulsen, Detlev Boison

Research output: Contribution to journalArticle

Abstract

Purpose: Given the high incidence of refractory epilepsy, novel therapeutic approaches and concepts are urgently needed. To date, viral-mediated delivery and endogenous expression of antisense sequences as a strategy to prevent seizures have received little attention in epilepsy therapy development efforts. Here we validate adenosine kinase (ADK), the astrocyte-based key negative regulator of the brain's endogenous anticonvulsant adenosine, as a potential therapeutic target for antisense-mediated seizure suppression. Methods: Wedeveloped adenoassociated virus 8 (AAV8)-based gene therapy vectors to selectively modulate ADK expression in astrocytes. Cell type selectivity was achieved by expressing an Adk-cDNA in sense or antisense orientation under the control of an astrocyte-specific gfaABC1D promoter. Viral vectors where injected into the CA3 of wild-type mice or spontaneously epileptic Adk-tg transgenic mice that overexpress ADK in brain. After virus injection, ADK expression was assessed histologically and biochemically. In addition, intracranial electroencephalography (EEG) recordings were obtained. Key Findings: We demonstrate in wild-type mice that viral overexpression of ADK within astrocytes is sufficient to trigger spontaneous recurrent seizures in the absence of any other epileptogenic event, whereas ADK down-regulation via AAV8-mediated RNA interference almost completely abolished spontaneous recurrent seizures in Adk-tg mice. Significance: Our data demonstrate that modulation of astrocytic ADK expression can trigger or prevent seizures, respectively. This is the first study to use an antisense approach to validate ADK as a rational therapeutic target for the treatment of epilepsy and suggests that gene therapies based on the knock down of ADK might be a feasible approach to control seizures in refractory epilepsy. Wiley Periodicals, Inc.

Original languageEnglish (US)
Pages (from-to)589-601
Number of pages13
JournalEpilepsia
Volume52
Issue number3
DOIs
StatePublished - Mar 1 2011
Externally publishedYes

Fingerprint

Adenosine Kinase
Epilepsy
Seizures
Astrocytes
Therapeutics
Viruses
Genetic Therapy
Absence Epilepsy
Brain
RNA Interference
Anticonvulsants
Adenosine
Transgenic Mice
Down-Regulation
Complementary DNA

Keywords

  • AAV8
  • Adenoassociated virus
  • ADK
  • Gene therapy
  • RNAi
  • Seizure

ASJC Scopus subject areas

  • Neurology
  • Clinical Neurology

Cite this

Theofilas, P., Brar, S., Mitchell, K-A., Shen, H. Y., Sandau, U. S., Poulsen, D., & Boison, D. (2011). Adenosine kinase as a target for therapeutic antisense strategies in epilepsy. Epilepsia, 52(3), 589-601. https://doi.org/10.1111/j.1528-1167.2010.02947.x

Adenosine kinase as a target for therapeutic antisense strategies in epilepsy. / Theofilas, Panos; Brar, Sukhmani; Mitchell, Kerry-Ann; Shen, Hai Ying; Sandau, Ursula S.; Poulsen, David; Boison, Detlev.

In: Epilepsia, Vol. 52, No. 3, 01.03.2011, p. 589-601.

Research output: Contribution to journalArticle

Theofilas, P, Brar, S, Mitchell, K-A, Shen, HY, Sandau, US, Poulsen, D & Boison, D 2011, 'Adenosine kinase as a target for therapeutic antisense strategies in epilepsy', Epilepsia, vol. 52, no. 3, pp. 589-601. https://doi.org/10.1111/j.1528-1167.2010.02947.x
Theofilas, Panos ; Brar, Sukhmani ; Mitchell, Kerry-Ann ; Shen, Hai Ying ; Sandau, Ursula S. ; Poulsen, David ; Boison, Detlev. / Adenosine kinase as a target for therapeutic antisense strategies in epilepsy. In: Epilepsia. 2011 ; Vol. 52, No. 3. pp. 589-601.
@article{3fedeacb4b204c30a9465bec24cdec5c,
title = "Adenosine kinase as a target for therapeutic antisense strategies in epilepsy",
abstract = "Purpose: Given the high incidence of refractory epilepsy, novel therapeutic approaches and concepts are urgently needed. To date, viral-mediated delivery and endogenous expression of antisense sequences as a strategy to prevent seizures have received little attention in epilepsy therapy development efforts. Here we validate adenosine kinase (ADK), the astrocyte-based key negative regulator of the brain's endogenous anticonvulsant adenosine, as a potential therapeutic target for antisense-mediated seizure suppression. Methods: Wedeveloped adenoassociated virus 8 (AAV8)-based gene therapy vectors to selectively modulate ADK expression in astrocytes. Cell type selectivity was achieved by expressing an Adk-cDNA in sense or antisense orientation under the control of an astrocyte-specific gfaABC1D promoter. Viral vectors where injected into the CA3 of wild-type mice or spontaneously epileptic Adk-tg transgenic mice that overexpress ADK in brain. After virus injection, ADK expression was assessed histologically and biochemically. In addition, intracranial electroencephalography (EEG) recordings were obtained. Key Findings: We demonstrate in wild-type mice that viral overexpression of ADK within astrocytes is sufficient to trigger spontaneous recurrent seizures in the absence of any other epileptogenic event, whereas ADK down-regulation via AAV8-mediated RNA interference almost completely abolished spontaneous recurrent seizures in Adk-tg mice. Significance: Our data demonstrate that modulation of astrocytic ADK expression can trigger or prevent seizures, respectively. This is the first study to use an antisense approach to validate ADK as a rational therapeutic target for the treatment of epilepsy and suggests that gene therapies based on the knock down of ADK might be a feasible approach to control seizures in refractory epilepsy. Wiley Periodicals, Inc.",
keywords = "AAV8, Adenoassociated virus, ADK, Gene therapy, RNAi, Seizure",
author = "Panos Theofilas and Sukhmani Brar and Kerry-Ann Mitchell and Shen, {Hai Ying} and Sandau, {Ursula S.} and David Poulsen and Detlev Boison",
year = "2011",
month = "3",
day = "1",
doi = "10.1111/j.1528-1167.2010.02947.x",
language = "English (US)",
volume = "52",
pages = "589--601",
journal = "Epilepsia",
issn = "0013-9580",
publisher = "Wiley-Blackwell",
number = "3",

}

TY - JOUR

T1 - Adenosine kinase as a target for therapeutic antisense strategies in epilepsy

AU - Theofilas, Panos

AU - Brar, Sukhmani

AU - Mitchell, Kerry-Ann

AU - Shen, Hai Ying

AU - Sandau, Ursula S.

AU - Poulsen, David

AU - Boison, Detlev

PY - 2011/3/1

Y1 - 2011/3/1

N2 - Purpose: Given the high incidence of refractory epilepsy, novel therapeutic approaches and concepts are urgently needed. To date, viral-mediated delivery and endogenous expression of antisense sequences as a strategy to prevent seizures have received little attention in epilepsy therapy development efforts. Here we validate adenosine kinase (ADK), the astrocyte-based key negative regulator of the brain's endogenous anticonvulsant adenosine, as a potential therapeutic target for antisense-mediated seizure suppression. Methods: Wedeveloped adenoassociated virus 8 (AAV8)-based gene therapy vectors to selectively modulate ADK expression in astrocytes. Cell type selectivity was achieved by expressing an Adk-cDNA in sense or antisense orientation under the control of an astrocyte-specific gfaABC1D promoter. Viral vectors where injected into the CA3 of wild-type mice or spontaneously epileptic Adk-tg transgenic mice that overexpress ADK in brain. After virus injection, ADK expression was assessed histologically and biochemically. In addition, intracranial electroencephalography (EEG) recordings were obtained. Key Findings: We demonstrate in wild-type mice that viral overexpression of ADK within astrocytes is sufficient to trigger spontaneous recurrent seizures in the absence of any other epileptogenic event, whereas ADK down-regulation via AAV8-mediated RNA interference almost completely abolished spontaneous recurrent seizures in Adk-tg mice. Significance: Our data demonstrate that modulation of astrocytic ADK expression can trigger or prevent seizures, respectively. This is the first study to use an antisense approach to validate ADK as a rational therapeutic target for the treatment of epilepsy and suggests that gene therapies based on the knock down of ADK might be a feasible approach to control seizures in refractory epilepsy. Wiley Periodicals, Inc.

AB - Purpose: Given the high incidence of refractory epilepsy, novel therapeutic approaches and concepts are urgently needed. To date, viral-mediated delivery and endogenous expression of antisense sequences as a strategy to prevent seizures have received little attention in epilepsy therapy development efforts. Here we validate adenosine kinase (ADK), the astrocyte-based key negative regulator of the brain's endogenous anticonvulsant adenosine, as a potential therapeutic target for antisense-mediated seizure suppression. Methods: Wedeveloped adenoassociated virus 8 (AAV8)-based gene therapy vectors to selectively modulate ADK expression in astrocytes. Cell type selectivity was achieved by expressing an Adk-cDNA in sense or antisense orientation under the control of an astrocyte-specific gfaABC1D promoter. Viral vectors where injected into the CA3 of wild-type mice or spontaneously epileptic Adk-tg transgenic mice that overexpress ADK in brain. After virus injection, ADK expression was assessed histologically and biochemically. In addition, intracranial electroencephalography (EEG) recordings were obtained. Key Findings: We demonstrate in wild-type mice that viral overexpression of ADK within astrocytes is sufficient to trigger spontaneous recurrent seizures in the absence of any other epileptogenic event, whereas ADK down-regulation via AAV8-mediated RNA interference almost completely abolished spontaneous recurrent seizures in Adk-tg mice. Significance: Our data demonstrate that modulation of astrocytic ADK expression can trigger or prevent seizures, respectively. This is the first study to use an antisense approach to validate ADK as a rational therapeutic target for the treatment of epilepsy and suggests that gene therapies based on the knock down of ADK might be a feasible approach to control seizures in refractory epilepsy. Wiley Periodicals, Inc.

KW - AAV8

KW - Adenoassociated virus

KW - ADK

KW - Gene therapy

KW - RNAi

KW - Seizure

UR - http://www.scopus.com/inward/record.url?scp=79952552797&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=79952552797&partnerID=8YFLogxK

U2 - 10.1111/j.1528-1167.2010.02947.x

DO - 10.1111/j.1528-1167.2010.02947.x

M3 - Article

C2 - 21275977

AN - SCOPUS:79952552797

VL - 52

SP - 589

EP - 601

JO - Epilepsia

JF - Epilepsia

SN - 0013-9580

IS - 3

ER -