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Detection of GNAQ mutations and reduction of cell viability in uveal melanoma cells with functionalized gold nanoparticles

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Abstract

Background

Uveal melanoma (UM) is the most common primary intraocular malignancy in adults. Early treatment may improve any chances of preventing metastatic disease, but diagnosis of small UM is challenging. Up to 95 % of all UMs carry somatic mutations in the G-coupled proteins GNAQ and GNA11 promoting anchorage-independent growth and proliferation. About 50 % of UMs are fatal. Once metastatic, patients have limited options for successful therapy.

Methods

We have developed functionalized gold nanoparticles (AuNPs) to visualize transcripts of mutant GNAQ mRNA in living cells. In addition to their suitability as a specific tool for GNAQ mutation detection, we have developed a novel linker that enables conjugation of siRNAs to AuNPs allowing for greater and more rapid intracellular release of siRNAs compared to previously described approaches.

Results

Binding of modified AuNPs to matching target mRNA leads to conformational changes, resulting in a detectable fluorescent signal that can be used for mutation detection in living cells. Knockdown of GNAQ with siRNA-AuNPs effectively reduced downstream signals and decreased cell viability in GNAQ mutant uveal melanoma cells.

Conclusion

AuNPs may in future be developed to serve as sensors for mutations of vital importance. The new release system for siRNA-AuNP improves previous systems, which conceivably will be useful for future therapeutic gene regulatory approaches.

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Acknowledgments

We thank Bertil Damato and James E. Cleaver for their helpful advice in the elaboration of this manuscript. We further thank Boris Bastian for providing the cell lines used in this study. This work was supported by, the National Institute of Health (1K08CA155035-01A1), the Melanoma Research Alliance, the UCSF Catalyst Program, the Max Kade Foundation, the Verein für Dermatologie Krankenanstalt Rudolfstiftung and the René Touraine Foundation the Ministry of Economy and competitiveness (SAF2010-15440) and IMDEA Nanociencia. We further acknowledge T. Dattels for his generous support.

Coflict of interests

The authors have no conflict of interest. Elements of the structure of gold nanoparticles used in this study are patented: 61/933,881 (US); EP14382033.0 (EU).

Author information

Authors and Affiliations

  1. Department of Dermatology, Mount Zion Cancer Research Center, University of California San Francisco, 2340 Sutter Street N461, 94115, San Francisco, CA, USA

    Christian Posch, Anna Celli, Igor Vujic, Martina Sanlorenzo, Gary A. Green, Jingly Weier, Mitchell Zekhtser, Jeffrey Ma, Gabriela Monico & Susana Ortiz-Urda

  2. Department of Dermatology, The Rudolfstiftung Hospital, Juchgasse 25, 1030, Vienna, Austria

    Christian Posch, Igor Vujic, Klemens Rappersberger & Álvaro Somoza

  3. Instituto Madrileño de Estudios Avanzados en Nanociencia (IMDEA Nanociencia), & CNB-CSIC-IMDEA Nanociencia Associated Unit “Unidad de Nanobiotecnología”, 28049, Madrid, Spain

    Alfonso Latorre & Ana Latorre

  4. North County Eye Surgery Associates, 320 Santa Fe Drive Suite 104, Encinitas, 92024, USA

    Michelle B. Crosby

  5. Department of Medical Sciences, Section of Dermatology, University of Turin, Via Cherasco 23, 10100, Torino, Italy

    Martina Sanlorenzo

  6. Departments of Ophthalmology, Stanford University, University of California San Francisco and California Pacific Medical Center, 45 Castro Street, 94114, San Francisco, USA

    Devron H. Char & Denis Jusufbegovic

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  1. Christian Posch
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  11. Jeffrey Ma
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  16. Álvaro Somoza
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  17. Susana Ortiz-Urda
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Corresponding author

Correspondence to Christian Posch.

Additional information

Christian Posch, Alfonso Latorre, Álvaro Somoza and Susana Ortiz-Urda contributed equally to this work.

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Posch, C., Latorre, A., Crosby, M.B. et al. Detection of GNAQ mutations and reduction of cell viability in uveal melanoma cells with functionalized gold nanoparticles. Biomed Microdevices 17, 15 (2015). https://doi.org/10.1007/s10544-014-9908-7

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