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AAV2-mediated CLN2 gene transfer to rodent and non-human primate brain results in long-term TPP-I expression compatible with therapy for LINCL

Gene Therapy volume 12, pages 1618–1632 (2005)Cite this article

Abstract

Late infantile neuronal ceroid lipofuscinosis (LINCL) is a fatal, autosomal recessive disease resulting from mutations in the CLN2 gene with consequent deficiency in its product tripeptidyl peptidase I (TPP-I). In the central nervous system (CNS), the deficiency of TPP-I results in the accumulation of proteins in lysosomes leading to a loss of neurons causing progressive neurological decline, and death by ages 10–12 years. To establish the feasibility of treating the CNS manifestations of LINCL by gene transfer, an adeno-associated virus 2 (AAV2) vector encoding the human CLN2 cDNA (AAV2CUhCLN2) was assessed for its ability to establish therapeutic levels of TPP-I in the brain. In vitro studies demonstrated that AAV2CUhCLN2 expressed CLN2 and produced biologically active TPP-I protein of which a fraction was secreted as the pro-TPP-I precursor and was taken up by nontransduced cells (ie, cross-correction). Following AAV2-mediated CLN2 delivery to the rat striatum, enzymatically active TPP-I protein was detected. By immunohistochemistry TPP-I protein was detected in striatal neurons (encompassing nearly half of the target structure) for up to 18 months. At the longer time points following striatal administration, TPP-I-positive cell bodies were also observed in the substantia nigra, frontal cerebral cortex and thalamus of the injected hemisphere, and the frontal cerebral cortex of the noninjected hemisphere. These areas of the brain contain neurons that extend axons into the striatum, suggesting that CNS circuitry may aid the distribution of the gene product. To assess the feasibility of human CNS delivery, a total of 3.6 × 1011 particle units of AAV2CUhCLN2 was administered to the CNS of African green monkeys in 12 distributed doses. Assessment at 5 and 13 weeks demonstrated widespread detection of TPP-I in neurons, but not glial cells, at all regions of injection. The distribution of TPP-I-positive cells was similar between the two time points at all injection sites. Together, these data support the development of direct CNS gene transfer using an AAV2 vector expressing the CLN2 cDNA for the CNS manifestations of LINCL.

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Acknowledgements

We thank P Lobel, Robert Wood Johnson Medical College, for providing the anti-TPP-I polyclonal antibody and the CLN2-deficient fibroblasts; K Wisniewski, Institute for Basic Research in Developmental Disabilities, for providing the anti-TPP-I monoclonal antibody; B Ferris, R McKinney, M Lam, B Bergman, J Qiu, E Salvin, E Vassallo, R Abplett, and the staff at the St Kitts Biomedical Research Foundation for technical assistance; and N Mohamed and T Virgin-Bryan for help in preparing this manuscript. These studies were supported, in part, by Nathan's Battle Foundation, Indianapolis, IN; and the Will Rogers Memorial Fund, Los Angeles, CA.

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  1. D Sondhi and D A Peterson: These authors contributed equally to this paper

Authors and Affiliations

  1. Department of Genetic Medicine, Weill Medical College of Cornell University, New York, NY, USA

    D Sondhi, E L Giannaris, B S Mendez, B De, P L Leopold, S M Kaminsky, N R Hackett & R G Crystal

  2. Department of Neuroscience, Rosalind Franklin University of Medicine and Science, The Chicago Medical School, North Chicago, IL, USA

    D A Peterson, C T Sanders & A B Rostkowski

  3. Department of Psychiatry, University of Colorado Health Science Center, Denver, CO, USA

    B Blanchard, K Bjugstad & J R Sladek Jr

  4. Neural Transplantation and Regeneration Program, School of Medicine, Yale University, New Haven, CT, USA

    D E Redmond Jr

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Sondhi, D., Peterson, D., Giannaris, E. et al. AAV2-mediated CLN2 gene transfer to rodent and non-human primate brain results in long-term TPP-I expression compatible with therapy for LINCL. Gene Ther 12, 1618–1632 (2005). https://doi.org/10.1038/sj.gt.3302549

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