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Cyclic stretch-induced reorganization of the cytoskeleton and its role in enhanced gene transfer

Gene Therapy volume 13, pages 725–731 (2006)Cite this article

Abstract

Cyclic stretch is known to alter a number of cellular and subcellular processes, including those involved in nonviral gene delivery. We have previously shown that moderate equibiaxial cyclic stretch (10% change in basement membrane area, 0.5 Hz, 50% duty cycle) of human pulmonary A549 cells enhances gene transfer and expression of reporter plasmid DNA in vitro, and that this phenomena may be due to alterations in cytoplasmic trafficking. Although the path by which plasmid DNA travels through the cytoplasm toward the nucleus is not well understood, the cytoskeleton and the constituents of the cytoplasm are known to significantly hinder macromolecular diffusion. Using biochemical techniques and immunofluorescence microscopy, we show that both the microfilament and microtubule networks are significantly reorganized by equibiaxial cyclic stretch. Prevention of this reorganization through the use of cytoskeletal stabilizing compounds mitigates the stretch-induced increase in gene expression, however, depolymerization in the absence of stretch is not sufficient to increase gene expression. These results suggest that cytoskeletal reorganization plays an important role in stretch-induced gene transfer and expression.

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Acknowledgements

We would like to thank Chris Capaccio, Joshua Gasiorowski, Erin Vaughan, and Jennifer Young for their helpful discussions and advice. This work was supported in part by Grants HL71643 (DAD and MRG) and HL076139 (RCG) from the NIH, and funds from the Institute for BioNanotechnology in Medicine of Northwestern University.

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Authors and Affiliations

  1. Division of Pulmonary and Critical Care Medicine, Northwestern University Medical School, Chicago, IL, USA

    R C Geiger, W Taylor & D A Dean

  2. Department of Biomedical Engineering, Northwestern University, Evanston, IL, USA

    M R Glucksberg

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  1. R C Geiger
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  2. W Taylor
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  3. M R Glucksberg
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  4. D A Dean
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Correspondence to D A Dean.

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Geiger, R., Taylor, W., Glucksberg, M. et al. Cyclic stretch-induced reorganization of the cytoskeleton and its role in enhanced gene transfer. Gene Ther 13, 725–731 (2006). https://doi.org/10.1038/sj.gt.3302693

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