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

43 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
Pages (from-to)409-420
Number of pages12
JournalJAMA Ophthalmology
Volume132
Issue number4
DOIs
StatePublished - Jan 1 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

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, 01.01.2014, 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{076400f0044d4f18a72c0a708a6ce088,
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",
month = "1",
day = "1",
doi = "10.1001/jamaophthalmol.2013.7630",
language = "English",
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 - 2014/1/1

Y1 - 2014/1/1

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

VL - 132

SP - 409

EP - 420

JO - JAMA Ophthalmology

JF - JAMA Ophthalmology

SN - 2168-6165

IS - 4

ER -