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Effect of siRNA pre-Exposure on Subsequent Response to siRNA Therapy

Pharmaceutical Research volume 32, pages 3813–3826 (2015)Cite this article

ABSTRACT

Purpose

An alternative cancer therapy based on RNA interference (RNAi) has shown considerable promise but the possibility of resistance development is not known. This study explored the possibility of therapeutic resistance against siRNA nanoparticles in human cancer cells.

Methods

Two approaches to siRNA treatment were undertaken using lipid-modified polyethylenimines, a single high concentration (shock) and repeated increasing concentrations (gradual). The targets were Mcl-1, RPS6KA5 and KSP in MDA-MB-435 cells.

Results

There was no evidence of resistance development in shock-treated cells, while the decrease in mRNA levels of targeted proteins was not as robust in naïve cells in gradual treatment. However, silencing efficiency was restored after a 7-day recovery period when expression of suppressed proteins returned to normal levels. Cellular uptake of siRNA was not affected by pre-treatments. Other mediators involved in cell survival and proliferation were altered in siRNA-treated cells, but only JUN silencing led to a heightened loss of viability. In vivo experiments demonstrated similar silencing efficiency at mRNA level after repeat doses.

Conclusions

Human cancer cells responded to repeat siRNA nanoparticles in a similar fashion after a temporary initial alteration and little, if any, resistance was evident against repeated siRNA treatments.

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Abbreviations

FAM:

Fluorescein amidite

KSP:

Kinesin spindle protein

Mcl-1:

Myeloid leukemia cell differentiation protein

mRNA:

Messenger RNA

NT:

Non-treated

PEI-LA:

Linoleic acid-substituted polyethylenimine

RISC:

RNA-induced silencing complex

RQ:

Relative quantity

RT-PCR:

Real-time polymerase chain reaction

siRNA:

Short interfering RNA

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ACKNOWLEDGMENTS AND DISCLOSURES

The authors would like to thank Mrs. Geraldine Barron for her invaluable help with development and optimization of confocal microscopy, and Dr. Robert Clarke (Georgetown University, Washington, DC) for providing the MDA-435 cells. This project was financially supported by a Breast Cancer Research Grant from Canadian Breast Cancer Foundation (CBCF) and a NSERC Discovery Grant.

Author information

Authors and Affiliations

  1. School of Pharmacy, Chapman University, #211, 9401 Jeronimo Road, Irvine, California, 92618, USA

    Hamidreza Montazeri Aliabadi

  2. Department of Chemical & Material Engineering, Faculty of Engineering, University of Alberta, #2-020 RTF, Edmonton, Alberta, T6G 2G6, Canada

    Hamidreza Montazeri Aliabadi, Parvin Mahdipoor, Cezary Kucharsky, Nicole Chan & Hasan Uludağ

  3. Faculty of Pharmacy and Pharmaceutical Sciences, University of Alberta, Edmonton, Alberta, T6G 2E1, Canada

    Hasan Uludağ

  4. Department of Biomedical Engineering, Faculty of Medicine and Dentistry, University of Alberta, Edmonton, Alberta, T6G 2R3, Canada

    Hasan Uludağ

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  1. Hamidreza Montazeri Aliabadi
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  2. Parvin Mahdipoor
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  4. Nicole Chan
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Correspondence to Hamidreza Montazeri Aliabadi or Hasan Uludağ.

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Montazeri Aliabadi, H., Mahdipoor, P., Kucharsky, C. et al. Effect of siRNA pre-Exposure on Subsequent Response to siRNA Therapy. Pharm Res 32, 3813–3826 (2015). https://doi.org/10.1007/s11095-015-1741-z

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  • DOI: https://doi.org/10.1007/s11095-015-1741-z

KEY WORDS

  • siRNA
  • cancer therapy
  • resistance
  • polymeric carriers