IS4-FAM, a fluorescent tool to study CXCR4 affinity and competitive antagonism in native cancer cells

Hamshaw, Isabel ORCID: https://orcid.org/0000-0001-6445-0738, Cominetti, Marco, Nana-Akyin, Princess, Ho Yee Ho, Ernie, Searcey, Mark ORCID: https://orcid.org/0000-0003-2273-8949 and Mueller, Anja ORCID: https://orcid.org/0000-0003-0774-0434 (2024) IS4-FAM, a fluorescent tool to study CXCR4 affinity and competitive antagonism in native cancer cells. Pharmacology Research and Perspectives, 12 (5). ISSN 2052-1707

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Abstract

The ability to accurately measure drug-target interaction is critical for the discovery of new therapeutics. Classical pharmacological bioassays such as radioligand or fluorescent ligand binding assays can define the affinity or Kd of a ligand for a receptor with the lower the Kd, the stronger the binding and the higher the affinity. However, in many drug discovery laboratories today, the target of interest if often artificially upregulated by means of transfection to modify the host cell's genetic makeup. This then potentially invalidates the assumptions of classical pharmacology affinity calculations as the receptor of interest is no longer at normal physiological densities. The CXCR4 receptor is expressed on many different cancer cell types and is associated with metastasis and poor prognosis. Therefore, the CXCR4 receptor is a desirable target for novel therapeutics. In this study, we explore the applicability of the newly developed fluorescently tagged CXCR4 antagonists, IS4-FAM as an investigative tool to study CXCR4 affinity and competitive antagonism in native, non-transfected cancer cells using confocal microscopy and flow cytometry. IS4-FAM directly labels CXCR4 in several cell lines including high CXCR4 expressing SK-MEL-28 (malignant melanoma) and PC3 (metastatic prostate cancer) and lower CXCR4 expressing THP-1 (acute monocytic leukemia) and was competitive with the established CXCR4 antagonist, AMD3100. This highlights the potential of IS4-FAM as a pharmacological tool for drug discovery in native cells lines and tissues.

Item Type:	Article
Additional Information:	Data availability statement: The datasets generated during and/or analyzed during the current study are available from the corresponding author on reasonable request. Funding information: UKRI Biotechnology and Biological Sciences Research Council Norwich Research Park Biosciences Doctoral Training Partnership. Grant Number: BB/T008717/1
Uncontrolled Keywords:	sdg 3 - good health and well-being ,/dk/atira/pure/sustainabledevelopmentgoals/good_health_and_well_being
Faculty \ School:	Faculty of Science > School of Chemistry, Pharmacy and Pharmacology
UEA Research Groups:	Faculty of Science > Research Groups > Molecular and Tissue Pharmacology
Depositing User:	LivePure Connector
Date Deposited:	20 Aug 2024 13:30
Last Modified:	02 Nov 2024 20:30
URI:	https://ueaeprints.uea.ac.uk/id/eprint/96283
DOI:	10.1002/prp2.70003

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