LHON gene therapy vector prevents visual loss and optic neuropathy induced by G11778A mutant mitochondrial DNA: Biodistribution and toxicology profile

Rajeshwari Koilkonda, Hong Yu, Venu Talla, Vittorio Porciatti, William J. Feuer, William W. Hauswirth, Vince Chiodo, Kirsten E. Erger, Sanford L. Boye, Alfred S. Lewin, Thomas J. Conlon, Lauren Renner, Martha Neuringer, Carol Detrisac, John Guy

    Research output: Contribution to journalArticle

    21 Citations (Scopus)

    Abstract

    Purpose: To demonstrate safety and efficacy of allotopic human ND4 for treatment of a Leber’s hereditary optic neuropathy (LHON) mouse model harboring the G11778A mitochondrial mutation.

    Methods: We induced LHON in mice by intravitreal injection of mutant (G11778A) human ND4 DNA, responsible for most cases of LHON, that was directed to mitochondria using an AAV2 vector to which we appended a mitochondrial targeting sequence to the VP2 capsid. We then attempted rescue of visual loss using our test article (ScAAV2-P1ND4v2) containing a synthetic nuclear encoded G11778G ND4 gene that was allotopically expressed. Control mice either were uninjected or received AAV2-GFP or AAV2-mCherry. We performed RT-PCR and confocal microscopy at 2 weeks post injection. Pattern electroretinograms (PERGs), spectraldomain optical coherence tomography (SD-OCT), histology, and transmission electron microscopy (TEM) were performed. For toxicology and biodistribution studies, the test article was administered intravitreally to rats and rhesus macaques at different doses.

    Results: Mutant and wild-type ND4 were efficiently expressed in the mitochondria of retinal ganglion cells (RGCs). Visual function assessed by serial PERGs and retinal structure by serial SD-OCT showed a significant rescue by the test article. Histology and ultrastructural analysis confirmed that loss of RGCs and demise of axons was prevented by ScAAV2-P1ND4v2. Rat and nonhuman primate biodistribution studies showed that vector spread outside the injected eye into spleen and lymph nodes was minimal. Histopathology of tissues and organs including the eyes was comparable to that of uninfected and saline-injected eyes.

    Conclusions: Allotopically expressed wild-type ND4 prevents the phenotype induced by G11778A mitochondrial DNA with a toxicology profile acceptable for testing in a phase I clinical trial.

    Original languageEnglish (US)
    Pages (from-to)7739-7753
    Number of pages15
    JournalInvestigative Ophthalmology and Visual Science
    Volume55
    Issue number12
    DOIs
    StatePublished - Oct 23 2014

    Fingerprint

    Leber's Hereditary Optic Atrophy
    Optic Nerve Diseases
    Mitochondrial DNA
    Genetic Therapy
    Toxicology
    Retinal Ganglion Cells
    Optical Coherence Tomography
    Histology
    Mitochondria
    Clinical Trials, Phase I
    Intravitreal Injections
    Capsid
    Macaca mulatta
    Transmission Electron Microscopy
    Confocal Microscopy
    Primates
    Axons
    Spleen
    Lymph Nodes
    Phenotype

    Keywords

    • Gene therapy
    • LHON
    • Mitochondria

    ASJC Scopus subject areas

    • Ophthalmology
    • Sensory Systems
    • Cellular and Molecular Neuroscience

    Cite this

    LHON gene therapy vector prevents visual loss and optic neuropathy induced by G11778A mutant mitochondrial DNA : Biodistribution and toxicology profile. / Koilkonda, Rajeshwari; Yu, Hong; Talla, Venu; Porciatti, Vittorio; Feuer, William J.; Hauswirth, William W.; Chiodo, Vince; Erger, Kirsten E.; Boye, Sanford L.; Lewin, Alfred S.; Conlon, Thomas J.; Renner, Lauren; Neuringer, Martha; Detrisac, Carol; Guy, John.

    In: Investigative Ophthalmology and Visual Science, Vol. 55, No. 12, 23.10.2014, p. 7739-7753.

    Research output: Contribution to journalArticle

    Koilkonda, R, Yu, H, Talla, V, Porciatti, V, Feuer, WJ, Hauswirth, WW, Chiodo, V, Erger, KE, Boye, SL, Lewin, AS, Conlon, TJ, Renner, L, Neuringer, M, Detrisac, C & Guy, J 2014, 'LHON gene therapy vector prevents visual loss and optic neuropathy induced by G11778A mutant mitochondrial DNA: Biodistribution and toxicology profile', Investigative Ophthalmology and Visual Science, vol. 55, no. 12, pp. 7739-7753. https://doi.org/10.1167/iovs.14-15388
    Koilkonda, Rajeshwari ; Yu, Hong ; Talla, Venu ; Porciatti, Vittorio ; Feuer, William J. ; Hauswirth, William W. ; Chiodo, Vince ; Erger, Kirsten E. ; Boye, Sanford L. ; Lewin, Alfred S. ; Conlon, Thomas J. ; Renner, Lauren ; Neuringer, Martha ; Detrisac, Carol ; Guy, John. / LHON gene therapy vector prevents visual loss and optic neuropathy induced by G11778A mutant mitochondrial DNA : Biodistribution and toxicology profile. In: Investigative Ophthalmology and Visual Science. 2014 ; Vol. 55, No. 12. pp. 7739-7753.
    @article{fd79150bd156471eb7b018726d916242,
    title = "LHON gene therapy vector prevents visual loss and optic neuropathy induced by G11778A mutant mitochondrial DNA: Biodistribution and toxicology profile",
    abstract = "Purpose: To demonstrate safety and efficacy of allotopic human ND4 for treatment of a Leber’s hereditary optic neuropathy (LHON) mouse model harboring the G11778A mitochondrial mutation.Methods: We induced LHON in mice by intravitreal injection of mutant (G11778A) human ND4 DNA, responsible for most cases of LHON, that was directed to mitochondria using an AAV2 vector to which we appended a mitochondrial targeting sequence to the VP2 capsid. We then attempted rescue of visual loss using our test article (ScAAV2-P1ND4v2) containing a synthetic nuclear encoded G11778G ND4 gene that was allotopically expressed. Control mice either were uninjected or received AAV2-GFP or AAV2-mCherry. We performed RT-PCR and confocal microscopy at 2 weeks post injection. Pattern electroretinograms (PERGs), spectraldomain optical coherence tomography (SD-OCT), histology, and transmission electron microscopy (TEM) were performed. For toxicology and biodistribution studies, the test article was administered intravitreally to rats and rhesus macaques at different doses.Results: Mutant and wild-type ND4 were efficiently expressed in the mitochondria of retinal ganglion cells (RGCs). Visual function assessed by serial PERGs and retinal structure by serial SD-OCT showed a significant rescue by the test article. Histology and ultrastructural analysis confirmed that loss of RGCs and demise of axons was prevented by ScAAV2-P1ND4v2. Rat and nonhuman primate biodistribution studies showed that vector spread outside the injected eye into spleen and lymph nodes was minimal. Histopathology of tissues and organs including the eyes was comparable to that of uninfected and saline-injected eyes.Conclusions: Allotopically expressed wild-type ND4 prevents the phenotype induced by G11778A mitochondrial DNA with a toxicology profile acceptable for testing in a phase I clinical trial.",
    keywords = "Gene therapy, LHON, Mitochondria",
    author = "Rajeshwari Koilkonda and Hong Yu and Venu Talla and Vittorio Porciatti and Feuer, {William J.} and Hauswirth, {William W.} and Vince Chiodo and Erger, {Kirsten E.} and Boye, {Sanford L.} and Lewin, {Alfred S.} and Conlon, {Thomas J.} and Lauren Renner and Martha Neuringer and Carol Detrisac and John Guy",
    year = "2014",
    month = "10",
    day = "23",
    doi = "10.1167/iovs.14-15388",
    language = "English (US)",
    volume = "55",
    pages = "7739--7753",
    journal = "Investigative Ophthalmology and Visual Science",
    issn = "0146-0404",
    publisher = "Association for Research in Vision and Ophthalmology Inc.",
    number = "12",

    }

    TY - JOUR

    T1 - LHON gene therapy vector prevents visual loss and optic neuropathy induced by G11778A mutant mitochondrial DNA

    T2 - Biodistribution and toxicology profile

    AU - Koilkonda, Rajeshwari

    AU - Yu, Hong

    AU - Talla, Venu

    AU - Porciatti, Vittorio

    AU - Feuer, William J.

    AU - Hauswirth, William W.

    AU - Chiodo, Vince

    AU - Erger, Kirsten E.

    AU - Boye, Sanford L.

    AU - Lewin, Alfred S.

    AU - Conlon, Thomas J.

    AU - Renner, Lauren

    AU - Neuringer, Martha

    AU - Detrisac, Carol

    AU - Guy, John

    PY - 2014/10/23

    Y1 - 2014/10/23

    N2 - Purpose: To demonstrate safety and efficacy of allotopic human ND4 for treatment of a Leber’s hereditary optic neuropathy (LHON) mouse model harboring the G11778A mitochondrial mutation.Methods: We induced LHON in mice by intravitreal injection of mutant (G11778A) human ND4 DNA, responsible for most cases of LHON, that was directed to mitochondria using an AAV2 vector to which we appended a mitochondrial targeting sequence to the VP2 capsid. We then attempted rescue of visual loss using our test article (ScAAV2-P1ND4v2) containing a synthetic nuclear encoded G11778G ND4 gene that was allotopically expressed. Control mice either were uninjected or received AAV2-GFP or AAV2-mCherry. We performed RT-PCR and confocal microscopy at 2 weeks post injection. Pattern electroretinograms (PERGs), spectraldomain optical coherence tomography (SD-OCT), histology, and transmission electron microscopy (TEM) were performed. For toxicology and biodistribution studies, the test article was administered intravitreally to rats and rhesus macaques at different doses.Results: Mutant and wild-type ND4 were efficiently expressed in the mitochondria of retinal ganglion cells (RGCs). Visual function assessed by serial PERGs and retinal structure by serial SD-OCT showed a significant rescue by the test article. Histology and ultrastructural analysis confirmed that loss of RGCs and demise of axons was prevented by ScAAV2-P1ND4v2. Rat and nonhuman primate biodistribution studies showed that vector spread outside the injected eye into spleen and lymph nodes was minimal. Histopathology of tissues and organs including the eyes was comparable to that of uninfected and saline-injected eyes.Conclusions: Allotopically expressed wild-type ND4 prevents the phenotype induced by G11778A mitochondrial DNA with a toxicology profile acceptable for testing in a phase I clinical trial.

    AB - Purpose: To demonstrate safety and efficacy of allotopic human ND4 for treatment of a Leber’s hereditary optic neuropathy (LHON) mouse model harboring the G11778A mitochondrial mutation.Methods: We induced LHON in mice by intravitreal injection of mutant (G11778A) human ND4 DNA, responsible for most cases of LHON, that was directed to mitochondria using an AAV2 vector to which we appended a mitochondrial targeting sequence to the VP2 capsid. We then attempted rescue of visual loss using our test article (ScAAV2-P1ND4v2) containing a synthetic nuclear encoded G11778G ND4 gene that was allotopically expressed. Control mice either were uninjected or received AAV2-GFP or AAV2-mCherry. We performed RT-PCR and confocal microscopy at 2 weeks post injection. Pattern electroretinograms (PERGs), spectraldomain optical coherence tomography (SD-OCT), histology, and transmission electron microscopy (TEM) were performed. For toxicology and biodistribution studies, the test article was administered intravitreally to rats and rhesus macaques at different doses.Results: Mutant and wild-type ND4 were efficiently expressed in the mitochondria of retinal ganglion cells (RGCs). Visual function assessed by serial PERGs and retinal structure by serial SD-OCT showed a significant rescue by the test article. Histology and ultrastructural analysis confirmed that loss of RGCs and demise of axons was prevented by ScAAV2-P1ND4v2. Rat and nonhuman primate biodistribution studies showed that vector spread outside the injected eye into spleen and lymph nodes was minimal. Histopathology of tissues and organs including the eyes was comparable to that of uninfected and saline-injected eyes.Conclusions: Allotopically expressed wild-type ND4 prevents the phenotype induced by G11778A mitochondrial DNA with a toxicology profile acceptable for testing in a phase I clinical trial.

    KW - Gene therapy

    KW - LHON

    KW - Mitochondria

    UR - http://www.scopus.com/inward/record.url?scp=84914166610&partnerID=8YFLogxK

    UR - http://www.scopus.com/inward/citedby.url?scp=84914166610&partnerID=8YFLogxK

    U2 - 10.1167/iovs.14-15388

    DO - 10.1167/iovs.14-15388

    M3 - Article

    C2 - 25342621

    AN - SCOPUS:84914166610

    VL - 55

    SP - 7739

    EP - 7753

    JO - Investigative Ophthalmology and Visual Science

    JF - Investigative Ophthalmology and Visual Science

    SN - 0146-0404

    IS - 12

    ER -