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Synchrotron phase-contrast X-ray imaging reveals fluid dosing dynamics for gene transfer into mouse airways

Gene Therapy volume 19, pages 8–14 (2012)Cite this article

Abstract

Although airway gene transfer research in mouse models relies on bolus fluid dosing into the nose or trachea, the dynamics and immediate fate of delivered gene transfer agents are poorly understood. In particular, this is because there are no in vivo methods able to accurately visualize the movement of fluid in small airways of intact animals. Using synchrotron phase-contrast X-ray imaging, we show that the fate of surrogate fluid doses delivered into live mouse airways can now be accurately and non-invasively monitored with high spatial and temporal resolution. This new imaging approach can help explain the non-homogenous distributions of gene expression observed in nasal airway gene transfer studies, suggests that substantial dose losses may occur at deliver into mouse trachea via immediate retrograde fluid motion and shows the influence of the speed of bolus delivery on the relative targeting of conducting and deeper lung airways. These findings provide insight into some of the factors that can influence gene expression in vivo, and this method provides a new approach to documenting and analyzing dose delivery in small-animal models.

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Acknowledgements

This study was supported by the NH&MRC Australia, with additional support from philanthropic donors via the CURE4CF Foundation (http://www.cure4cf.org). The synchrotron radiation experiments were performed on the BL20B2 beamline at SPring-8, with the approval of the Japan Synchrotron Radiation Institute (JASRI) under proposal number 2010A1523. Professor Naoto Yagi and Dr Kentaro Uesugi provided expert synchrotron imaging and controls advice and assistance at the SPring-8 synchrotron. We thank P Cmielewski, M Limberis and D Miller for their editorial input. MD, KS, AJ and DP were supported by the International Synchrotron Access Program (ISAP) managed by the Australian Synchrotron. The ISAP is an initiative of the Australian Government being conducted as part of the National Collaborative Research Infrastructure Strategy.

Author contributions

DP conceived the research; MD and DP devised the experiments; MD and DP performed the animal experiments in conjunction with KS and AJ who performed the synchrotron imaging; MD and DP conceived the image analyses; MD performed the image analyses and constructed the figures and Supplementary Material; MD and DP wrote the initial manuscript and all authors edited and approved the final version.

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Authors and Affiliations

  1. Department of Respiratory and Sleep Medicine, Women's and Children's Hospital, Adelaide, Australia

    M Donnelley & D W Parsons

  2. Centre for Stem Cell Research, University of Adelaide, Adelaide, Australia

    M Donnelley & D W Parsons

  3. Department of Paediatrics and Reproductive Health, University of Adelaide, Adelaide, Australia

    M Donnelley & D W Parsons

  4. Monash Biomedical Imaging, Monash University, Clayton, Victoria, Australia

    K K W Siu

  5. School of Physics, Monash University, Clayton, Victoria, Australia

    K K W Siu

  6. Australian Synchrotron, Clayton, Victoria, Australia

    K K W Siu

  7. Division of Biological Engineering, Monash University, Monash, Victoria, Australia

    R A Jamison

  8. Women's and Children's Health Research Institute, Adelaide, Australia

    D W Parsons

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  1. M Donnelley
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  2. K K W Siu
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Correspondence to M Donnelley.

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Donnelley, M., Siu, K., Jamison, R. et al. Synchrotron phase-contrast X-ray imaging reveals fluid dosing dynamics for gene transfer into mouse airways. Gene Ther 19, 8–14 (2012). https://doi.org/10.1038/gt.2011.80

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