Mutant sodium channel for tumor therapy

Bakhos A. Tannous; Adam P. Christensen; Lisa Pike; Thomas Wurdinger; Katherine F. Perry; Okay Saydam; Andreas H. Jacobs; Jaime García-Añoveros; Ralph Weissleder; Miguel Sena-Esteves; David P. Corey; Xandra O. Breakefield

doi:10.1038/mt.2009.33

Mutant sodium channel for tumor therapy

Bakhos A. Tannous^*, Adam P. Christensen, Lisa Pike, Thomas Wurdinger, Katherine F. Perry, Okay Saydam, Andreas H. Jacobs, Jaime García-Añoveros, Ralph Weissleder, Miguel Sena-Esteves, David P. Corey, Xandra O. Breakefield

^*Corresponding author for this work

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

17 Scopus citations

Abstract

Viral vectors have been used to deliver a wide range of therapeutic genes to tumors. In this study, a novel tumor therapy was achieved by the delivery of a mammalian brain sodium channel, ASIC2a, carrying a mutation that renders it constitutively open. This channel was delivered to tumor cells using a herpes simplex virus-1/Epstein-Barr virus (HSV/ EBV) hybrid amplicon vector in which gene expression was controlled by a tetracycline regulatory system (tet-on) with silencer elements. Upon infection and doxycycline induction of mutant channel expression in tumor cells, the open channel led to amiloride-sensitive sodium influx as assessed by patch clamp recording and sodium imaging in culture. Within hours, tumor cells swelled and died. In addition to cells expressing the mutant channel, adjacent, noninfected cells connected by gap junctions also died. Intratumoral injection of HSV/EBV amplicon vector encoding the mutant sodium channel and systemic administration of doxycycline led to regression of subcutaneous tumors in nude mice as assessed by in vivo bioluminescence imaging. The advantage of this direct mode of tumor therapy is that all types of tumor cells become susceptible and death is rapid with no time for the tumor cells to become resistant.

Original language	English (US)
Pages (from-to)	810-819
Number of pages	10
Journal	Molecular Therapy
Volume	17
Issue number	5
DOIs	https://doi.org/10.1038/mt.2009.33
State	Published - 2009

Funding

We thank Dr Bujard for providing tetracycline regulatory elements used in the amplicon vector and Dr Fraefel for providing the HSV-TK amplicon plasmid; Dr Rachamimov (Pathologist at MGH) for analyzing the hematoxylin and eosin stained sections. Nicole Lewandrowski, Juliet Fernandez, Ozlem Senol, and Johanna M. Niers for technical help; Dr Yellen for access to the sodium imaging microscope; Suzanne McDavitt for skilled editorial assistance; and Igor Bagayev for assistance with confocal microscopy. This work was supported by grants from the National Cancer Institute (CA69246 and CA86355 to XOB and RW, and 1K99CA126839 to BAT), from NINDS (NS44363 to JG-A) and from the Brain Tumor Society (to B.A.T. and X.O.B.). D.P.C. is an Investigator of the Howard Hughes Medical Institute.

ASJC Scopus subject areas

Drug Discovery
Genetics
Molecular Medicine
Molecular Biology
Pharmacology

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10.1038/mt.2009.33

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title = "Mutant sodium channel for tumor therapy",

abstract = "Viral vectors have been used to deliver a wide range of therapeutic genes to tumors. In this study, a novel tumor therapy was achieved by the delivery of a mammalian brain sodium channel, ASIC2a, carrying a mutation that renders it constitutively open. This channel was delivered to tumor cells using a herpes simplex virus-1/Epstein-Barr virus (HSV/ EBV) hybrid amplicon vector in which gene expression was controlled by a tetracycline regulatory system (tet-on) with silencer elements. Upon infection and doxycycline induction of mutant channel expression in tumor cells, the open channel led to amiloride-sensitive sodium influx as assessed by patch clamp recording and sodium imaging in culture. Within hours, tumor cells swelled and died. In addition to cells expressing the mutant channel, adjacent, noninfected cells connected by gap junctions also died. Intratumoral injection of HSV/EBV amplicon vector encoding the mutant sodium channel and systemic administration of doxycycline led to regression of subcutaneous tumors in nude mice as assessed by in vivo bioluminescence imaging. The advantage of this direct mode of tumor therapy is that all types of tumor cells become susceptible and death is rapid with no time for the tumor cells to become resistant.",

author = "Tannous, {Bakhos A.} and Christensen, {Adam P.} and Lisa Pike and Thomas Wurdinger and Perry, {Katherine F.} and Okay Saydam and Jacobs, {Andreas H.} and Jaime Garc{\'i}a-A{\~n}overos and Ralph Weissleder and Miguel Sena-Esteves and Corey, {David P.} and Breakefield, {Xandra O.}",

note = "Funding Information: We thank Dr Bujard for providing tetracycline regulatory elements used in the amplicon vector and Dr Fraefel for providing the HSV-TK amplicon plasmid; Dr Rachamimov (Pathologist at MGH) for analyzing the hematoxylin and eosin stained sections. Nicole Lewandrowski, Juliet Fernandez, Ozlem Senol, and Johanna M. Niers for technical help; Dr Yellen for access to the sodium imaging microscope; Suzanne McDavitt for skilled editorial assistance; and Igor Bagayev for assistance with confocal microscopy. This work was supported by grants from the National Cancer Institute (CA69246 and CA86355 to XOB and RW, and 1K99CA126839 to BAT), from NINDS (NS44363 to JG-A) and from the Brain Tumor Society (to B.A.T. and X.O.B.). D.P.C. is an Investigator of the Howard Hughes Medical Institute.",

year = "2009",

doi = "10.1038/mt.2009.33",

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AU - Christensen, Adam P.

AU - Pike, Lisa

AU - Wurdinger, Thomas

AU - Perry, Katherine F.

AU - Saydam, Okay

AU - Jacobs, Andreas H.

AU - García-Añoveros, Jaime

AU - Weissleder, Ralph

AU - Sena-Esteves, Miguel

AU - Corey, David P.

AU - Breakefield, Xandra O.

N1 - Funding Information: We thank Dr Bujard for providing tetracycline regulatory elements used in the amplicon vector and Dr Fraefel for providing the HSV-TK amplicon plasmid; Dr Rachamimov (Pathologist at MGH) for analyzing the hematoxylin and eosin stained sections. Nicole Lewandrowski, Juliet Fernandez, Ozlem Senol, and Johanna M. Niers for technical help; Dr Yellen for access to the sodium imaging microscope; Suzanne McDavitt for skilled editorial assistance; and Igor Bagayev for assistance with confocal microscopy. This work was supported by grants from the National Cancer Institute (CA69246 and CA86355 to XOB and RW, and 1K99CA126839 to BAT), from NINDS (NS44363 to JG-A) and from the Brain Tumor Society (to B.A.T. and X.O.B.). D.P.C. is an Investigator of the Howard Hughes Medical Institute.

PY - 2009

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N2 - Viral vectors have been used to deliver a wide range of therapeutic genes to tumors. In this study, a novel tumor therapy was achieved by the delivery of a mammalian brain sodium channel, ASIC2a, carrying a mutation that renders it constitutively open. This channel was delivered to tumor cells using a herpes simplex virus-1/Epstein-Barr virus (HSV/ EBV) hybrid amplicon vector in which gene expression was controlled by a tetracycline regulatory system (tet-on) with silencer elements. Upon infection and doxycycline induction of mutant channel expression in tumor cells, the open channel led to amiloride-sensitive sodium influx as assessed by patch clamp recording and sodium imaging in culture. Within hours, tumor cells swelled and died. In addition to cells expressing the mutant channel, adjacent, noninfected cells connected by gap junctions also died. Intratumoral injection of HSV/EBV amplicon vector encoding the mutant sodium channel and systemic administration of doxycycline led to regression of subcutaneous tumors in nude mice as assessed by in vivo bioluminescence imaging. The advantage of this direct mode of tumor therapy is that all types of tumor cells become susceptible and death is rapid with no time for the tumor cells to become resistant.

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Mutant sodium channel for tumor therapy

Abstract

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ASJC Scopus subject areas

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