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

Christian Posch, Alfonso Latorre, Michelle B. Crosby, Anna Celli, Ana Latorre, Igor Vujic, Martina Sanlorenzo, Gary A. Green, Jingly Weier, Mitchell Zekhtser, Jeffrey Ma, Gabriela Monico, Devron H. Char, Denis Jusufbegovic, Klemens Rappersberger, Álvaro Somoza, Susana Ortiz-Urda

Research output: Contribution to journalArticle

9 Citations (Scopus)

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.

Original languageEnglish (US)
JournalBiomedical Microdevices
Volume17
Issue number1
DOIs
StatePublished - Jan 1 2015
Externally publishedYes

Fingerprint

Gold
Nanoparticles
Cell Survival
Cells
Mutation
Small Interfering RNA
Messenger RNA
Regulator Genes
Genes
Proteins
Therapeutics
Sensors
Uveal melanoma
Growth
Neoplasms

Keywords

  • Fluorescence
  • GNAQ
  • Linker
  • siRNA
  • Treatment

ASJC Scopus subject areas

  • Biomedical Engineering
  • Molecular Biology

Cite this

Detection of GNAQ mutations and reduction of cell viability in uveal melanoma cells with functionalized gold nanoparticles. / Posch, Christian; Latorre, Alfonso; Crosby, Michelle B.; Celli, Anna; Latorre, Ana; Vujic, Igor; Sanlorenzo, Martina; Green, Gary A.; Weier, Jingly; Zekhtser, Mitchell; Ma, Jeffrey; Monico, Gabriela; Char, Devron H.; Jusufbegovic, Denis; Rappersberger, Klemens; Somoza, Álvaro; Ortiz-Urda, Susana.

In: Biomedical Microdevices, Vol. 17, No. 1, 01.01.2015.

Research output: Contribution to journalArticle

Posch, C, Latorre, A, Crosby, MB, Celli, A, Latorre, A, Vujic, I, Sanlorenzo, M, Green, GA, Weier, J, Zekhtser, M, Ma, J, Monico, G, Char, DH, Jusufbegovic, D, Rappersberger, K, Somoza, Á & Ortiz-Urda, S 2015, 'Detection of GNAQ mutations and reduction of cell viability in uveal melanoma cells with functionalized gold nanoparticles', Biomedical Microdevices, vol. 17, no. 1. https://doi.org/10.1007/s10544-014-9908-7
Posch, Christian ; Latorre, Alfonso ; Crosby, Michelle B. ; Celli, Anna ; Latorre, Ana ; Vujic, Igor ; Sanlorenzo, Martina ; Green, Gary A. ; Weier, Jingly ; Zekhtser, Mitchell ; Ma, Jeffrey ; Monico, Gabriela ; Char, Devron H. ; Jusufbegovic, Denis ; Rappersberger, Klemens ; Somoza, Álvaro ; Ortiz-Urda, Susana. / Detection of GNAQ mutations and reduction of cell viability in uveal melanoma cells with functionalized gold nanoparticles. In: Biomedical Microdevices. 2015 ; Vol. 17, No. 1.
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AU - Latorre, Ana

AU - Vujic, Igor

AU - Sanlorenzo, Martina

AU - Green, Gary A.

AU - Weier, Jingly

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AU - Ma, Jeffrey

AU - Monico, Gabriela

AU - Char, Devron H.

AU - Jusufbegovic, Denis

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AU - Somoza, Álvaro

AU - Ortiz-Urda, Susana

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AB - 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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