Cited by 670Adenosine and Purinergic SignalingBiochemical and Molecular Researchbioluminescence and chemiluminescence research

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Increased Level of Extracellular ATP at Tumor Sites: In Vivo Imaging with Plasma Membrane Luciferase

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Pellegatti, Patrizia, Raffaghello, Lizzia, Bianchi, Giovanna, Piccardi, Federica, Pistoia, Vito, Di Virgilio, Francesco·2008·PLoS ONE

DOI 10.1371/journal.pone.0002599PMC PMC2440522

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Abstract

BACKGROUND: There is growing awareness that tumour cells build up a "self-advantageous" microenvironment that reduces effectiveness of anti-tumour immune response. While many different immunosuppressive mechanisms are likely to come into play, recent evidence suggests that extracellular adenosine acting at A2A receptors may have a major role in down-modulating the immune response as cancerous tissues contain elevated levels of adenosine and adenosine break-down products. While there is no doubt that all cells possess plasma membrane adenosine transporters that mediate adenosine uptake and may also allow its release, it is now clear that most of extracellularly-generated adenosine originates from the catabolism of extracellular ATP. METHODOLOGY/PRINCIPAL FINDINGS: Measurement of extracellular ATP is generally performed in cell supernatants by HPLC or soluble luciferin-luciferase assay, thus it generally turns out to be laborious and inaccurate. We have engineered a chimeric plasma membrane-targeted luciferase that allows in vivo real-time imaging of extracellular ATP. With this novel probe we have measured the ATP concentration within the tumour microenvironment of several experimentally-induced tumours. CONCLUSIONS/SIGNIFICANCE: Our results show that ATP in the tumour interstitium is in the hundreds micromolar range, while it is basically undetectable in healthy tissues. Here we show that a chimeric plasma membrane-targeted luciferase allows in vivo detection of high extracellular ATP concentration at tumour sites. On the contrary, tumour-free tissues show undetectable extracellular ATP levels. Extracellular ATP may be crucial for the tumour not only as a stimulus for growth but also as a source of an immunosuppressive agent such as adenosine. Our approach offers a new tool for the investigation of the biochemical composition of tumour milieu and for development of novel therapies based on the modulation of extracellular purine-based signalling.

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Authors: Pellegatti, P., Raffaghello, L., Bianchi, G., Piccardi, F., Pistoia, V., & Di Virgilio, F.
Affiliations: University of Ferrara (IT)
Istituto Giannina Gaslini (IT)
ORCID: Raffaghello, Lizzia
Bianchi, Giovanna
Pistoia, Vito
Venue: PLoS ONE
Publisher: Public Library of Science (PLoS)
Published: 2008-07-09
Volume: 3
Issue: 7
Pages: e2599
ISSN: 1932-6203 (ISSN-L)
DOI: 10.1371/journal.pone.0002599
PMC: PMC2440522
OpenAlex: W2070663123
Cited by: 668
References: 28
Citations by year: 2026: 31 · 2025: 40 · 2024: 29 · 2023: 62 · 2022: 51 · 2021: 58 · 2020: 66 · 2019: 42 · 2018: 37 · 2017: 34 · 2016: 33 · 2015: 25 · 2014: 36 · 2013: 28 · 2012: 40
FWCI: 11.40
Type: journal article
Language: en
Open access: gold
doi.org
Licence: cc-by
creativecommons.org/licenses/by/4.0/ (unspecified)
Locations: PLoS ONE (publishedVersion, cc-by)
PubMed Central (publishedVersion, cc-by)
PubMed (publishedVersion)
DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
Institutional Research Information System University of Ferrara (University of Ferrara) (submittedVersion)
Funders: Fondazione Italiana per la Lotta al Neuroblastoma
Associazione Italiana per la Ricerca sul Cancro (GGP06070)
Università degli Studi di Ferrara
Topics: Life Sciences › Biochemistry, Genetics and Molecular Biology › Physiology › Adenosine and Purinergic Signaling
Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Biochemical and Molecular Research
Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › bioluminescence and chemiluminescence research
Keywords: extracellular ATP, tumor microenvironment, in vivo imaging, adenosine, adenosine A2A receptor, immunosuppression, ATP catabolism, adenosine transporter
MeSH terms: Adenosine Triphosphate, Animals, Cell Membrane, Extracellular Space, Humans, Luciferases, Luminescent Measurements, Mice, Nude, Neoplasms, Cell Line, Tumor, Luminescent Agents, Mice

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Increased Level of Extracellular ATP at Tumor Sites: In Vivo Imaging with Plasma Membrane Luciferase

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