Molecularly targeted nanoparticles: an emerging tool for evaluation of expression of the receptor for advanced glycation end products in a murine model of peripheral artery disease

Abstract Background Molecular imaging with molecularly targeted probes is a powerful tool for studying the spatio-temporal interactions between complex biological processes. The pivotal role of the receptor for advanced glycation end products (RAGE), and its involvement in numerous pathological proc...

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Main Authors: Marcin Woźniak, Christian J. Konopka, Agata Płoska, Jamila Hedhli, Anna Siekierzycka, Maciej Banach, Rafal Bartoszewski, Lawrence W. Dobrucki, Leszek Kalinowski, Iwona T. Dobrucki
Format: Article
Language:English
Published: BMC 2021-03-01
Series:Cellular & Molecular Biology Letters
Subjects:
Online Access:https://doi.org/10.1186/s11658-021-00253-0
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spelling doaj-b4a7f210e06e43ae8bd35ad11a59b45f2021-03-21T12:20:37ZengBMCCellular & Molecular Biology Letters1425-81531689-13922021-03-0126111710.1186/s11658-021-00253-0Molecularly targeted nanoparticles: an emerging tool for evaluation of expression of the receptor for advanced glycation end products in a murine model of peripheral artery diseaseMarcin Woźniak0Christian J. Konopka1Agata Płoska2Jamila Hedhli3Anna Siekierzycka4Maciej Banach5Rafal Bartoszewski6Lawrence W. Dobrucki7Leszek Kalinowski8Iwona T. Dobrucki9Department of Medical Laboratory Diagnostics – Fahrenheit Biobank BBMRI.pl, Medical University of GdanskUniversity of Illinois at Urbana-Champaign Beckman Institute for Advanced Science and TechnologyDepartment of Medical Laboratory Diagnostics – Fahrenheit Biobank BBMRI.pl, Medical University of GdanskUniversity of Illinois at Urbana-Champaign Beckman Institute for Advanced Science and TechnologyDepartment of Medical Laboratory Diagnostics – Fahrenheit Biobank BBMRI.pl, Medical University of GdanskDepartment of Hypertension, Medical University of LodzDepartment of Biology and Pharmaceutical Botany, Medical University of GdanskDepartment of Medical Laboratory Diagnostics – Fahrenheit Biobank BBMRI.pl, Medical University of GdanskDepartment of Medical Laboratory Diagnostics – Fahrenheit Biobank BBMRI.pl, Medical University of GdanskUniversity of Illinois at Urbana-Champaign Beckman Institute for Advanced Science and TechnologyAbstract Background Molecular imaging with molecularly targeted probes is a powerful tool for studying the spatio-temporal interactions between complex biological processes. The pivotal role of the receptor for advanced glycation end products (RAGE), and its involvement in numerous pathological processes, aroused the demand for RAGE-targeted imaging in various diseases. In the present study, we evaluated the use of a diagnostic imaging agent for RAGE quantification in an animal model of peripheral artery disease, a multimodal dual-labeled probe targeted at RAGE (MMIA-CML). Methods PAMAM dendrimer was conjugated with Nε-carboxymethyl-lysine (CML) modified albumin to synthesize the RAGE-targeted probe. A control untargeted agent carried native non-modified human albumin (HSA). Bifunctional p-SCN-Bn-NOTA was used to conjugate the 64Cu radioisotope. Surgical right femoral artery ligation was performed on C57BL/6 male mice. One week after femoral artery ligation, mice were injected with MMIA-CML or MMIA-HSA labeled with 64Cu radioisotope and 60 min later in vivo microPET-CT imaging was performed. Immediately after PET imaging studies, the murine hindlimb muscle tissues were excised and prepared for gene and protein expression analysis. RAGE gene and protein expression was assessed using real-time qPCR and Western blot technique respectively. To visualize RAGE expression in excised tissues, microscopic fluorescence imaging was performed using RAGE-specific antibodies and RAGE-targeted and -control MMIA. Results Animals subjected to PET imaging exhibited greater MMIA-CML uptake in ischemic hindlimbs than non-ischemic hindlimbs. We observed a high correlation between fluorescent signal detection and radioactivity measurement. Significant RAGE gene and protein overexpression were observed in ischemic hindlimbs compared to non-ischemic hindlimbs at one week after surgical ligation. Fluorescence microscopic staining revealed significantly increased uptake of RAGE-targeted nanoparticles in both ischemic and non-ischemic muscle tissues compared to the control probe but at a higher level in ischemic hindlimbs. Ischemic tissue exhibited explicit RAGE dyeing following anti-RAGE antibody and high colocalization with the MMIA-CML targeted at RAGE. Conclusions The present results indicate increased expression of RAGE in the ischemic hindlimb and enable the use of multimodal nanoparticles in both in vitro and in vivo experimental models, creating the possibility for imaging structural and functional changes with a RAGE-targeted tracer.https://doi.org/10.1186/s11658-021-00253-0RAGEAGEsMolecular imagingIschemia
collection DOAJ
language English
format Article
sources DOAJ
author Marcin Woźniak
Christian J. Konopka
Agata Płoska
Jamila Hedhli
Anna Siekierzycka
Maciej Banach
Rafal Bartoszewski
Lawrence W. Dobrucki
Leszek Kalinowski
Iwona T. Dobrucki
spellingShingle Marcin Woźniak
Christian J. Konopka
Agata Płoska
Jamila Hedhli
Anna Siekierzycka
Maciej Banach
Rafal Bartoszewski
Lawrence W. Dobrucki
Leszek Kalinowski
Iwona T. Dobrucki
Molecularly targeted nanoparticles: an emerging tool for evaluation of expression of the receptor for advanced glycation end products in a murine model of peripheral artery disease
Cellular & Molecular Biology Letters
RAGE
AGEs
Molecular imaging
Ischemia
author_facet Marcin Woźniak
Christian J. Konopka
Agata Płoska
Jamila Hedhli
Anna Siekierzycka
Maciej Banach
Rafal Bartoszewski
Lawrence W. Dobrucki
Leszek Kalinowski
Iwona T. Dobrucki
author_sort Marcin Woźniak
title Molecularly targeted nanoparticles: an emerging tool for evaluation of expression of the receptor for advanced glycation end products in a murine model of peripheral artery disease
title_short Molecularly targeted nanoparticles: an emerging tool for evaluation of expression of the receptor for advanced glycation end products in a murine model of peripheral artery disease
title_full Molecularly targeted nanoparticles: an emerging tool for evaluation of expression of the receptor for advanced glycation end products in a murine model of peripheral artery disease
title_fullStr Molecularly targeted nanoparticles: an emerging tool for evaluation of expression of the receptor for advanced glycation end products in a murine model of peripheral artery disease
title_full_unstemmed Molecularly targeted nanoparticles: an emerging tool for evaluation of expression of the receptor for advanced glycation end products in a murine model of peripheral artery disease
title_sort molecularly targeted nanoparticles: an emerging tool for evaluation of expression of the receptor for advanced glycation end products in a murine model of peripheral artery disease
publisher BMC
series Cellular & Molecular Biology Letters
issn 1425-8153
1689-1392
publishDate 2021-03-01
description Abstract Background Molecular imaging with molecularly targeted probes is a powerful tool for studying the spatio-temporal interactions between complex biological processes. The pivotal role of the receptor for advanced glycation end products (RAGE), and its involvement in numerous pathological processes, aroused the demand for RAGE-targeted imaging in various diseases. In the present study, we evaluated the use of a diagnostic imaging agent for RAGE quantification in an animal model of peripheral artery disease, a multimodal dual-labeled probe targeted at RAGE (MMIA-CML). Methods PAMAM dendrimer was conjugated with Nε-carboxymethyl-lysine (CML) modified albumin to synthesize the RAGE-targeted probe. A control untargeted agent carried native non-modified human albumin (HSA). Bifunctional p-SCN-Bn-NOTA was used to conjugate the 64Cu radioisotope. Surgical right femoral artery ligation was performed on C57BL/6 male mice. One week after femoral artery ligation, mice were injected with MMIA-CML or MMIA-HSA labeled with 64Cu radioisotope and 60 min later in vivo microPET-CT imaging was performed. Immediately after PET imaging studies, the murine hindlimb muscle tissues were excised and prepared for gene and protein expression analysis. RAGE gene and protein expression was assessed using real-time qPCR and Western blot technique respectively. To visualize RAGE expression in excised tissues, microscopic fluorescence imaging was performed using RAGE-specific antibodies and RAGE-targeted and -control MMIA. Results Animals subjected to PET imaging exhibited greater MMIA-CML uptake in ischemic hindlimbs than non-ischemic hindlimbs. We observed a high correlation between fluorescent signal detection and radioactivity measurement. Significant RAGE gene and protein overexpression were observed in ischemic hindlimbs compared to non-ischemic hindlimbs at one week after surgical ligation. Fluorescence microscopic staining revealed significantly increased uptake of RAGE-targeted nanoparticles in both ischemic and non-ischemic muscle tissues compared to the control probe but at a higher level in ischemic hindlimbs. Ischemic tissue exhibited explicit RAGE dyeing following anti-RAGE antibody and high colocalization with the MMIA-CML targeted at RAGE. Conclusions The present results indicate increased expression of RAGE in the ischemic hindlimb and enable the use of multimodal nanoparticles in both in vitro and in vivo experimental models, creating the possibility for imaging structural and functional changes with a RAGE-targeted tracer.
topic RAGE
AGEs
Molecular imaging
Ischemia
url https://doi.org/10.1186/s11658-021-00253-0
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