Safety and effects of the vector for the leber hereditary optic neuropathy gene therapy clinical trial

Rajeshwari D. Koilkonda, Hong Yu, Tsung Han Chou, William J. Feuer, Marco Ruggeri, Vittorio Porciatti, David Tse, William W. Hauswirth, Vince Chiodo, Sanford L. Boye, Alfred S. Lewin, Martha Neuringer, Lauren Renner, John Guy

    Research output: Contribution to journalArticle

    44 Citations (Scopus)

    Abstract

    IMPORTANCE We developed a novel strategy for treatment of Leber hereditary optic neuropathy (LHON) caused by a mutation in the nicotinamide adenine dinucleotide dehydrogenase subunit IV (ND4), mitochondrial gene. OBJECTIVE To demonstrate the safety and effects of the gene therapy vector to be used in a proposed gene therapy clinical trial. DESIGN AND SETTING In a series of laboratory experiments, we modified the mitochondrial ND4 subunit of complex I in the nuclear genetic code for import into mitochondria. The protein was targeted into the organelle by agency of a targeting sequence (allotopic expression). The gene was packaged into adeno-associated viral vectors and then vitreally injected into rodent, nonhuman primate, and ex vivo human eyes that underwent testing for expression and integration by immunohistochemical analysis and blue native polyacrylamide gel electrophoresis. During serial follow-up, the animal eyes underwent fundus photography, optical coherence tomography, and multifocal or pattern electroretinography.We tested for rescue of visual loss in rodent eyes also injected with a mutant G11778A ND4 homologue responsible for most cases of LHON. EXPOSURE Ocular infection with recombinant adeno-associated viral vectors containing a wild-type allotopic human ND4 gene. MAIN OUTCOMES AND MEASURES Expression of human ND4 and rescue of optic neuropathy induced by mutant human ND4. RESULTS We found human ND4 expressed in almost all mouse retinal ganglion cells by 1 week after injection and ND4 integrated into the mouse complex I. In rodent eyes also injected with a mutant allotopic ND4, wild-type allotopic ND4 prevented defective adenosine triphosphate synthesis, suppressed visual loss, reduced apoptosis of retinal ganglion cells, and prevented demise of axons in the optic nerve. Injection of ND4 in the ex vivo human eye resulted in expression in most retinal ganglion cells. Primates undergoing vitreal injection with the ND4 test article and followed up for 3 months had no serious adverse reactions. CONCLUSIONS AND RELEVANCE Expression of our allotopic ND4 vector in the ex vivo human eye, safety of the test article, rescue of the LHON mouse model, and the severe irreversible loss of visual function in LHON support clinical testing with mutated G11778A mitochondrial DNA in our patients.

    Original languageEnglish (US)
    Pages (from-to)409-420
    Number of pages12
    JournalJAMA Ophthalmology
    Volume132
    Issue number4
    DOIs
    StatePublished - 2014

    Fingerprint

    Leber's Hereditary Optic Atrophy
    Genetic Therapy
    Clinical Trials
    Safety
    Retinal Ganglion Cells
    Rodentia
    Primates
    Injections
    Genetic Code
    Electroretinography
    Native Polyacrylamide Gel Electrophoresis
    Eye Infections
    Optic Nerve Diseases
    Mitochondrial Genes
    Photography
    Optical Coherence Tomography
    Optic Nerve
    Mitochondrial DNA
    Organelles
    NAD

    ASJC Scopus subject areas

    • Ophthalmology

    Cite this

    Koilkonda, R. D., Yu, H., Chou, T. H., Feuer, W. J., Ruggeri, M., Porciatti, V., ... Guy, J. (2014). Safety and effects of the vector for the leber hereditary optic neuropathy gene therapy clinical trial. JAMA Ophthalmology, 132(4), 409-420. https://doi.org/10.1001/jamaophthalmol.2013.7630

    Safety and effects of the vector for the leber hereditary optic neuropathy gene therapy clinical trial. / Koilkonda, Rajeshwari D.; Yu, Hong; Chou, Tsung Han; Feuer, William J.; Ruggeri, Marco; Porciatti, Vittorio; Tse, David; Hauswirth, William W.; Chiodo, Vince; Boye, Sanford L.; Lewin, Alfred S.; Neuringer, Martha; Renner, Lauren; Guy, John.

    In: JAMA Ophthalmology, Vol. 132, No. 4, 2014, p. 409-420.

    Research output: Contribution to journalArticle

    Koilkonda, RD, Yu, H, Chou, TH, Feuer, WJ, Ruggeri, M, Porciatti, V, Tse, D, Hauswirth, WW, Chiodo, V, Boye, SL, Lewin, AS, Neuringer, M, Renner, L & Guy, J 2014, 'Safety and effects of the vector for the leber hereditary optic neuropathy gene therapy clinical trial', JAMA Ophthalmology, vol. 132, no. 4, pp. 409-420. https://doi.org/10.1001/jamaophthalmol.2013.7630
    Koilkonda, Rajeshwari D. ; Yu, Hong ; Chou, Tsung Han ; Feuer, William J. ; Ruggeri, Marco ; Porciatti, Vittorio ; Tse, David ; Hauswirth, William W. ; Chiodo, Vince ; Boye, Sanford L. ; Lewin, Alfred S. ; Neuringer, Martha ; Renner, Lauren ; Guy, John. / Safety and effects of the vector for the leber hereditary optic neuropathy gene therapy clinical trial. In: JAMA Ophthalmology. 2014 ; Vol. 132, No. 4. pp. 409-420.
    @article{8943f8440fe049e9bdcafb671b97f19f,
    title = "Safety and effects of the vector for the leber hereditary optic neuropathy gene therapy clinical trial",
    abstract = "IMPORTANCE We developed a novel strategy for treatment of Leber hereditary optic neuropathy (LHON) caused by a mutation in the nicotinamide adenine dinucleotide dehydrogenase subunit IV (ND4), mitochondrial gene. OBJECTIVE To demonstrate the safety and effects of the gene therapy vector to be used in a proposed gene therapy clinical trial. DESIGN AND SETTING In a series of laboratory experiments, we modified the mitochondrial ND4 subunit of complex I in the nuclear genetic code for import into mitochondria. The protein was targeted into the organelle by agency of a targeting sequence (allotopic expression). The gene was packaged into adeno-associated viral vectors and then vitreally injected into rodent, nonhuman primate, and ex vivo human eyes that underwent testing for expression and integration by immunohistochemical analysis and blue native polyacrylamide gel electrophoresis. During serial follow-up, the animal eyes underwent fundus photography, optical coherence tomography, and multifocal or pattern electroretinography.We tested for rescue of visual loss in rodent eyes also injected with a mutant G11778A ND4 homologue responsible for most cases of LHON. EXPOSURE Ocular infection with recombinant adeno-associated viral vectors containing a wild-type allotopic human ND4 gene. MAIN OUTCOMES AND MEASURES Expression of human ND4 and rescue of optic neuropathy induced by mutant human ND4. RESULTS We found human ND4 expressed in almost all mouse retinal ganglion cells by 1 week after injection and ND4 integrated into the mouse complex I. In rodent eyes also injected with a mutant allotopic ND4, wild-type allotopic ND4 prevented defective adenosine triphosphate synthesis, suppressed visual loss, reduced apoptosis of retinal ganglion cells, and prevented demise of axons in the optic nerve. Injection of ND4 in the ex vivo human eye resulted in expression in most retinal ganglion cells. Primates undergoing vitreal injection with the ND4 test article and followed up for 3 months had no serious adverse reactions. CONCLUSIONS AND RELEVANCE Expression of our allotopic ND4 vector in the ex vivo human eye, safety of the test article, rescue of the LHON mouse model, and the severe irreversible loss of visual function in LHON support clinical testing with mutated G11778A mitochondrial DNA in our patients.",
    author = "Koilkonda, {Rajeshwari D.} and Hong Yu and Chou, {Tsung Han} and Feuer, {William J.} and Marco Ruggeri and Vittorio Porciatti and David Tse and Hauswirth, {William W.} and Vince Chiodo and Boye, {Sanford L.} and Lewin, {Alfred S.} and Martha Neuringer and Lauren Renner and John Guy",
    year = "2014",
    doi = "10.1001/jamaophthalmol.2013.7630",
    language = "English (US)",
    volume = "132",
    pages = "409--420",
    journal = "JAMA Ophthalmology",
    issn = "2168-6165",
    publisher = "American Medical Association",
    number = "4",

    }

    TY - JOUR

    T1 - Safety and effects of the vector for the leber hereditary optic neuropathy gene therapy clinical trial

    AU - Koilkonda, Rajeshwari D.

    AU - Yu, Hong

    AU - Chou, Tsung Han

    AU - Feuer, William J.

    AU - Ruggeri, Marco

    AU - Porciatti, Vittorio

    AU - Tse, David

    AU - Hauswirth, William W.

    AU - Chiodo, Vince

    AU - Boye, Sanford L.

    AU - Lewin, Alfred S.

    AU - Neuringer, Martha

    AU - Renner, Lauren

    AU - Guy, John

    PY - 2014

    Y1 - 2014

    N2 - IMPORTANCE We developed a novel strategy for treatment of Leber hereditary optic neuropathy (LHON) caused by a mutation in the nicotinamide adenine dinucleotide dehydrogenase subunit IV (ND4), mitochondrial gene. OBJECTIVE To demonstrate the safety and effects of the gene therapy vector to be used in a proposed gene therapy clinical trial. DESIGN AND SETTING In a series of laboratory experiments, we modified the mitochondrial ND4 subunit of complex I in the nuclear genetic code for import into mitochondria. The protein was targeted into the organelle by agency of a targeting sequence (allotopic expression). The gene was packaged into adeno-associated viral vectors and then vitreally injected into rodent, nonhuman primate, and ex vivo human eyes that underwent testing for expression and integration by immunohistochemical analysis and blue native polyacrylamide gel electrophoresis. During serial follow-up, the animal eyes underwent fundus photography, optical coherence tomography, and multifocal or pattern electroretinography.We tested for rescue of visual loss in rodent eyes also injected with a mutant G11778A ND4 homologue responsible for most cases of LHON. EXPOSURE Ocular infection with recombinant adeno-associated viral vectors containing a wild-type allotopic human ND4 gene. MAIN OUTCOMES AND MEASURES Expression of human ND4 and rescue of optic neuropathy induced by mutant human ND4. RESULTS We found human ND4 expressed in almost all mouse retinal ganglion cells by 1 week after injection and ND4 integrated into the mouse complex I. In rodent eyes also injected with a mutant allotopic ND4, wild-type allotopic ND4 prevented defective adenosine triphosphate synthesis, suppressed visual loss, reduced apoptosis of retinal ganglion cells, and prevented demise of axons in the optic nerve. Injection of ND4 in the ex vivo human eye resulted in expression in most retinal ganglion cells. Primates undergoing vitreal injection with the ND4 test article and followed up for 3 months had no serious adverse reactions. CONCLUSIONS AND RELEVANCE Expression of our allotopic ND4 vector in the ex vivo human eye, safety of the test article, rescue of the LHON mouse model, and the severe irreversible loss of visual function in LHON support clinical testing with mutated G11778A mitochondrial DNA in our patients.

    AB - IMPORTANCE We developed a novel strategy for treatment of Leber hereditary optic neuropathy (LHON) caused by a mutation in the nicotinamide adenine dinucleotide dehydrogenase subunit IV (ND4), mitochondrial gene. OBJECTIVE To demonstrate the safety and effects of the gene therapy vector to be used in a proposed gene therapy clinical trial. DESIGN AND SETTING In a series of laboratory experiments, we modified the mitochondrial ND4 subunit of complex I in the nuclear genetic code for import into mitochondria. The protein was targeted into the organelle by agency of a targeting sequence (allotopic expression). The gene was packaged into adeno-associated viral vectors and then vitreally injected into rodent, nonhuman primate, and ex vivo human eyes that underwent testing for expression and integration by immunohistochemical analysis and blue native polyacrylamide gel electrophoresis. During serial follow-up, the animal eyes underwent fundus photography, optical coherence tomography, and multifocal or pattern electroretinography.We tested for rescue of visual loss in rodent eyes also injected with a mutant G11778A ND4 homologue responsible for most cases of LHON. EXPOSURE Ocular infection with recombinant adeno-associated viral vectors containing a wild-type allotopic human ND4 gene. MAIN OUTCOMES AND MEASURES Expression of human ND4 and rescue of optic neuropathy induced by mutant human ND4. RESULTS We found human ND4 expressed in almost all mouse retinal ganglion cells by 1 week after injection and ND4 integrated into the mouse complex I. In rodent eyes also injected with a mutant allotopic ND4, wild-type allotopic ND4 prevented defective adenosine triphosphate synthesis, suppressed visual loss, reduced apoptosis of retinal ganglion cells, and prevented demise of axons in the optic nerve. Injection of ND4 in the ex vivo human eye resulted in expression in most retinal ganglion cells. Primates undergoing vitreal injection with the ND4 test article and followed up for 3 months had no serious adverse reactions. CONCLUSIONS AND RELEVANCE Expression of our allotopic ND4 vector in the ex vivo human eye, safety of the test article, rescue of the LHON mouse model, and the severe irreversible loss of visual function in LHON support clinical testing with mutated G11778A mitochondrial DNA in our patients.

    UR - http://www.scopus.com/inward/record.url?scp=84898640526&partnerID=8YFLogxK

    UR - http://www.scopus.com/inward/citedby.url?scp=84898640526&partnerID=8YFLogxK

    U2 - 10.1001/jamaophthalmol.2013.7630

    DO - 10.1001/jamaophthalmol.2013.7630

    M3 - Article

    C2 - 24457989

    AN - SCOPUS:84898640526

    VL - 132

    SP - 409

    EP - 420

    JO - JAMA Ophthalmology

    JF - JAMA Ophthalmology

    SN - 2168-6165

    IS - 4

    ER -