Cargando…

CPPs to the Test: Effects on Binding, Uptake and Biodistribution of a Tumor Targeting Nanobody

Nanobodies are well-established targeting ligands for molecular imaging and therapy. Their short circulation time enables early imaging and reduces systemic radiation exposure. However, shorter circulation time leads to lower tracer accumulation in the target tissue. Cell-penetrating peptides (CPPs)...

Descripción completa

Detalles Bibliográficos
Autores principales: Collado Camps, Estel, van Lith, Sanne A. M., Frielink, Cathelijne, Lankhof, Jordi, Dijkgraaf, Ingrid, Gotthardt, Martin, Brock, Roland
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8308549/
https://www.ncbi.nlm.nih.gov/pubmed/34201507
http://dx.doi.org/10.3390/ph14070602
_version_ 1783728307327467520
author Collado Camps, Estel
van Lith, Sanne A. M.
Frielink, Cathelijne
Lankhof, Jordi
Dijkgraaf, Ingrid
Gotthardt, Martin
Brock, Roland
author_facet Collado Camps, Estel
van Lith, Sanne A. M.
Frielink, Cathelijne
Lankhof, Jordi
Dijkgraaf, Ingrid
Gotthardt, Martin
Brock, Roland
author_sort Collado Camps, Estel
collection PubMed
description Nanobodies are well-established targeting ligands for molecular imaging and therapy. Their short circulation time enables early imaging and reduces systemic radiation exposure. However, shorter circulation time leads to lower tracer accumulation in the target tissue. Cell-penetrating peptides (CPPs) improve cellular uptake of various cargoes, including nanobodies. CPPs could enhance tissue retention without compromising rapid clearance. However, systematic investigations on how the functionalities of nanobody and CPP combine with each other at the level of 2D and 3D cell cultures and in vivo are lacking. Here, we demonstrate that conjugates of the epidermal growth factor receptor (EGFR)-binding nanobody 7D12 with different CPPs (nonaarginine, penetratin, Tat and hLF) differ with respect to cell binding and induction of endocytosis. For nonaarginine and penetratin we compared the competition of EGF binding and performance of L- and D-peptide stereoisomers, and tested the D-peptide conjugates in tumor cell spheroids and in vivo. The D-peptide conjugates showed better penetration into spheroids than the unconjugated 7D12. Both in vivo and in vitro, the behavior of the agent reflects the combination of both functionalities. Although CPPs cause promising increases in in vitro uptake and 3D penetration, the dominant effect of the CPP in the control of biodistribution warrants further investigation.
format Online
Article
Text
id pubmed-8308549
institution National Center for Biotechnology Information
language English
publishDate 2021
publisher MDPI
record_format MEDLINE/PubMed
spelling pubmed-83085492021-07-25 CPPs to the Test: Effects on Binding, Uptake and Biodistribution of a Tumor Targeting Nanobody Collado Camps, Estel van Lith, Sanne A. M. Frielink, Cathelijne Lankhof, Jordi Dijkgraaf, Ingrid Gotthardt, Martin Brock, Roland Pharmaceuticals (Basel) Article Nanobodies are well-established targeting ligands for molecular imaging and therapy. Their short circulation time enables early imaging and reduces systemic radiation exposure. However, shorter circulation time leads to lower tracer accumulation in the target tissue. Cell-penetrating peptides (CPPs) improve cellular uptake of various cargoes, including nanobodies. CPPs could enhance tissue retention without compromising rapid clearance. However, systematic investigations on how the functionalities of nanobody and CPP combine with each other at the level of 2D and 3D cell cultures and in vivo are lacking. Here, we demonstrate that conjugates of the epidermal growth factor receptor (EGFR)-binding nanobody 7D12 with different CPPs (nonaarginine, penetratin, Tat and hLF) differ with respect to cell binding and induction of endocytosis. For nonaarginine and penetratin we compared the competition of EGF binding and performance of L- and D-peptide stereoisomers, and tested the D-peptide conjugates in tumor cell spheroids and in vivo. The D-peptide conjugates showed better penetration into spheroids than the unconjugated 7D12. Both in vivo and in vitro, the behavior of the agent reflects the combination of both functionalities. Although CPPs cause promising increases in in vitro uptake and 3D penetration, the dominant effect of the CPP in the control of biodistribution warrants further investigation. MDPI 2021-06-23 /pmc/articles/PMC8308549/ /pubmed/34201507 http://dx.doi.org/10.3390/ph14070602 Text en © 2021 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Collado Camps, Estel
van Lith, Sanne A. M.
Frielink, Cathelijne
Lankhof, Jordi
Dijkgraaf, Ingrid
Gotthardt, Martin
Brock, Roland
CPPs to the Test: Effects on Binding, Uptake and Biodistribution of a Tumor Targeting Nanobody
title CPPs to the Test: Effects on Binding, Uptake and Biodistribution of a Tumor Targeting Nanobody
title_full CPPs to the Test: Effects on Binding, Uptake and Biodistribution of a Tumor Targeting Nanobody
title_fullStr CPPs to the Test: Effects on Binding, Uptake and Biodistribution of a Tumor Targeting Nanobody
title_full_unstemmed CPPs to the Test: Effects on Binding, Uptake and Biodistribution of a Tumor Targeting Nanobody
title_short CPPs to the Test: Effects on Binding, Uptake and Biodistribution of a Tumor Targeting Nanobody
title_sort cpps to the test: effects on binding, uptake and biodistribution of a tumor targeting nanobody
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8308549/
https://www.ncbi.nlm.nih.gov/pubmed/34201507
http://dx.doi.org/10.3390/ph14070602
work_keys_str_mv AT colladocampsestel cppstothetesteffectsonbindinguptakeandbiodistributionofatumortargetingnanobody
AT vanlithsanneam cppstothetesteffectsonbindinguptakeandbiodistributionofatumortargetingnanobody
AT frielinkcathelijne cppstothetesteffectsonbindinguptakeandbiodistributionofatumortargetingnanobody
AT lankhofjordi cppstothetesteffectsonbindinguptakeandbiodistributionofatumortargetingnanobody
AT dijkgraafingrid cppstothetesteffectsonbindinguptakeandbiodistributionofatumortargetingnanobody
AT gotthardtmartin cppstothetesteffectsonbindinguptakeandbiodistributionofatumortargetingnanobody
AT brockroland cppstothetesteffectsonbindinguptakeandbiodistributionofatumortargetingnanobody