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)...
Autores principales: | , , , , , , |
---|---|
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 |