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Original Article – Enabling Technologies

Noninvasive tracking of gene transcript and neuroprotection after gene therapy

Gene Therapy volume 23, pages 1–9 (2016)Cite this article

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Abstract

Gene therapy holds exceptional potential for translational medicine by improving the products of defective genes in diseases and/or providing necessary biologics from endogenous sources during recovery processes. However, validating methods for the delivery, distribution and expression of the exogenous genes from such therapy can generally not be applicable to monitor effects over the long term because they are invasive. We report here that human granulocyte colony-stimulating factor (hG-CSF) complimentary DNA (cDNA) encoded in self-complementary adeno-associated virus-type 2 adeno-associated virus, as delivered through eye drops at multiple time points after cerebral ischemia using bilateral carotid occlusion for 60 min (BCAO-60) led to significant reduction in mortality rates, cerebral atrophy and neurological deficits in C57black6 mice. Most importantly, we validated hG-CSF cDNA expression using translatable magnetic resonance imaging (MRI) in living brains. This noninvasive approach for monitoring exogenous gene expression in the brains has potential for great impact in the area of experimental gene therapy in animal models of heart attack, stroke, Alzheimer’s dementia, Parkinson’s disorder and amyotrophic lateral sclerosis, and the translation of such techniques to emergency medicine.

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Acknowledgements

We thank Dr Sean I Savitz (University of Texas Houston Medical School, Houston, TX, USA) for demonstration of the corner tests; Dr Jarek Aronowski (UTHMS, TX, USA) for sharing the hypothermia treatment protocol; Dr Charng-Ming Liu for synthesizing all sODN and Dr JS Yang for conducting real time-PCR. This project was supported by NIH grants R01DA029889 and R01EB013768, as well as by a pilot grant from the Boston Area Diabetes Endocrinology Research Center (P30DK057521-14 (J Avruch)) to PKL, NS045776 to the MGH Neuroscience Center). The MR systems used for this work were funded by NIH Shared Instrumentation Grants (1S10RR025563) awarded to the Athinoula A Martinos Center for Biomedical Imaging. The B Wellcome Trust (No.1012722) to HP, State of Florida (09KW-11) to JYW also supported this research.

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Authors and Affiliations

  1. Department of Radiology, Athinoula A. Martinos Center for Biomedical Imaging, Massachusetts General Hospital and the Harvard Medical School, Charlestown, MA, USA

    J Ren, Y I Chen, C H Liu & P K Liu

  2. Department of Biomedical Science, Charles E. Schmidt College of Medicine, Florida Atlantic University, Boca Raton, FL, USA

    P-C Chen, H Prentice & J-Y Wu

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  1. J Ren
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Correspondence to P K Liu.

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Ren, J., Chen, Y., Liu, C. et al. Noninvasive tracking of gene transcript and neuroprotection after gene therapy. Gene Ther 23, 1–9 (2016). https://doi.org/10.1038/gt.2015.81

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