Monitoring extracellular ion and metabolite dynamics with recombinant nanobody-fused biosensors.

Autor:	Burgstaller S; Pharmaceutical Biotechnology, Eberhard Karls University Tübingen, Markwiesenstrasse 55, 72770 Reutlingen, Germany.; Department of Pharmacology, Toxicology and Clinical Pharmacy, Institute of Pharmacy, University of Tübingen, Auf der Morgenstelle 8, 72076 Tuebingen, Germany., Wagner TR; Pharmaceutical Biotechnology, Eberhard Karls University Tübingen, Markwiesenstrasse 55, 72770 Reutlingen, Germany.; NMI Natural and Medical Sciences Institute at the University of Tübingen, Markwiesenstrasse 55, 72770 Reutlingen, Germany., Bischof H; Department of Pharmacology, Toxicology and Clinical Pharmacy, Institute of Pharmacy, University of Tübingen, Auf der Morgenstelle 8, 72076 Tuebingen, Germany., Bueckle S; Pharmaceutical Biotechnology, Eberhard Karls University Tübingen, Markwiesenstrasse 55, 72770 Reutlingen, Germany., Padamsey A; Pharmaceutical Biotechnology, Eberhard Karls University Tübingen, Markwiesenstrasse 55, 72770 Reutlingen, Germany., Frecot D; Pharmaceutical Biotechnology, Eberhard Karls University Tübingen, Markwiesenstrasse 55, 72770 Reutlingen, Germany.; NMI Natural and Medical Sciences Institute at the University of Tübingen, Markwiesenstrasse 55, 72770 Reutlingen, Germany., Kaiser PD; NMI Natural and Medical Sciences Institute at the University of Tübingen, Markwiesenstrasse 55, 72770 Reutlingen, Germany., Skrabak D; Department of Pharmacology, Toxicology and Clinical Pharmacy, Institute of Pharmacy, University of Tübingen, Auf der Morgenstelle 8, 72076 Tuebingen, Germany., Malli R; Gottfried Schatz Research Center, Molecular Biology and Biochemistry, Medical University of Graz, Neue Stiftingtalstraße 6/6, 8010 Graz, Austria.; BioTechMed-Graz, Mozartgasse 12/II, 8010 Graz, Austria., Lukowski R; Department of Pharmacology, Toxicology and Clinical Pharmacy, Institute of Pharmacy, University of Tübingen, Auf der Morgenstelle 8, 72076 Tuebingen, Germany., Rothbauer U; Pharmaceutical Biotechnology, Eberhard Karls University Tübingen, Markwiesenstrasse 55, 72770 Reutlingen, Germany.; NMI Natural and Medical Sciences Institute at the University of Tübingen, Markwiesenstrasse 55, 72770 Reutlingen, Germany.
Jazyk:	angličtina
Zdroj:	IScience [iScience] 2022 Aug 10; Vol. 25 (9), pp. 104907. Date of Electronic Publication: 2022 Aug 10 (Print Publication: 2022).
DOI:	10.1016/j.isci.2022.104907
Abstrakt:	Ion and analyte changes in the tumor microenvironment (TME) alter the metabolic activity of cancer cells, promote tumor cell growth, and impair anti-tumor immunity. Consequently, accurate determination and visualization of extracellular changes of analytes in real time is desired. In this study, we genetically combined FRET-based biosensors with nanobodies (Nbs) to specifically visualize and monitor extracellular changes in K + , pH, and glucose on cell surfaces. We demonstrated that these Nb-fused biosensors quantitatively visualized K + alterations on cancer and non-cancer cell lines and primary neurons. By implementing a HER2-specific Nb, we generated functional K + and pH sensors, which specifically stained HER2-positive breast cancer cells. Based on the successful development of several Nb-fused biosensor combinations, we anticipate that this approach can be readily extended to other biosensors and will open new opportunities for the study of extracellular analytes in advanced experimental settings. Competing Interests: U.R. is named as inventor on a patent application (PCT/EP2016/077785) claiming the use of the described SPOT-Nb for research and diagnosis filed by the Natural and Medical Sciences Institute and ChromoTek. H.B. and R.M. are named as inventors on a patent application (US2018328908) claiming the use of the GEPII 1.0 for research and diagnosis filed by the Medical University of Graz. U.R. is a scientific advisor of the company ChromoTek which provided the sequence of the SPOT-Nb. All other authors declare no competing financial interest. (© 2022 The Author(s).)
Databáze:	MEDLINE
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