In vitro and ex vivo evaluation of preclinical models for FAP-targeted theranostics: differences and relevance for radiotracer evaluation

Abstract Background Fibroblast activation protein (FAP) is an attractive target for cancer theranostics. Although FAP-targeted nuclear imaging demonstrated promising clinical results, only sub-optimal results are reported for targeted radionuclide therapy (TRT). Preclinical research is crucial in se...

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Main Authors: Circe D. van der Heide, Joana D. Campeiro, Eline A. M. Ruigrok, Lilian van den Brink, Shashikanth Ponnala, Shawn M. Hillier, Simone U. Dalm
Format: Article
Language:English
Published: SpringerOpen 2024-12-01
Series:EJNMMI Research
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Online Access:https://doi.org/10.1186/s13550-024-01191-6
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author Circe D. van der Heide
Joana D. Campeiro
Eline A. M. Ruigrok
Lilian van den Brink
Shashikanth Ponnala
Shawn M. Hillier
Simone U. Dalm
author_facet Circe D. van der Heide
Joana D. Campeiro
Eline A. M. Ruigrok
Lilian van den Brink
Shashikanth Ponnala
Shawn M. Hillier
Simone U. Dalm
author_sort Circe D. van der Heide
collection DOAJ
description Abstract Background Fibroblast activation protein (FAP) is an attractive target for cancer theranostics. Although FAP-targeted nuclear imaging demonstrated promising clinical results, only sub-optimal results are reported for targeted radionuclide therapy (TRT). Preclinical research is crucial in selecting promising FAP-targeted radiopharmaceuticals and for obtaining an increased understanding of factors essential for FAP-TRT improvement. FAP is mainly expressed by cancer-associated fibroblasts in the tumor stroma and less on cancer cells themselves. Therefore, other (complex) factors impact FAP-TRT efficacy compared to currently clinically applied TRT strategies. For accurate evaluation of these aspects, selection of a representative preclinical model is important. Currently mainly human cancer cell lines transduced to (over)express FAP are applied, lacking clinical representation. It is unclear how these and more physiological FAP-expressing models compare to each other, and whether/how the model influences the study outcome. We aimed to address this by comparing FAP tracer behavior in FAP-transduced HT1080-huFAP and HEK293-huFAP cells, and endogenous FAP-expressing U-87 MG cancer cells and PS-1 pancreatic stellate cells. [111In]In-FAPI-46 and a fluorescent FAP-targeted tracer (RTX-1370S) were used to compare tracer binding/uptake and localization in vitro and ex vivo. Additionally, FAP expression was determined with RT-qPCR and anti-FAP IHC. Results Although FAP expression was highest in HEK293-huFAP cells and cell line derived xenografts, this did not result in the highest tracer uptake. [111In]In-FAPI-46 uptake was highest in HT1080-huFAP, closely followed by HEK293-huFAP, and a 6-10-fold lower uptake for U-87 MG and PS-1 cells. However, ex vivo U-87 MG xenografts only showed a 2-fold lower binding compared to HT1080-huFAP and HEK293-huFAP xenografts, mainly because the cell line attracts murine fibroblasts as demonstrated in our RT-qPCR and IHC studies. Conclusions The interaction between FAP and FAP-targeted tracers differs between models, indicating the need for appropriate model selection and that comparing results across studies using different models is difficult.
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spelling doaj-art-3c9a4964d6cd44f694ffdf3cbf59bb8e2025-08-20T02:43:26ZengSpringerOpenEJNMMI Research2191-219X2024-12-0114111310.1186/s13550-024-01191-6In vitro and ex vivo evaluation of preclinical models for FAP-targeted theranostics: differences and relevance for radiotracer evaluationCirce D. van der Heide0Joana D. Campeiro1Eline A. M. Ruigrok2Lilian van den Brink3Shashikanth Ponnala4Shawn M. Hillier5Simone U. Dalm6Department of Radiology & Nuclear Medicine, Erasmus MC University Medical Centre RotterdamDepartment of Radiology & Nuclear Medicine, Erasmus MC University Medical Centre RotterdamDepartment of Radiology & Nuclear Medicine, Erasmus MC University Medical Centre RotterdamDepartment of Radiology & Nuclear Medicine, Erasmus MC University Medical Centre RotterdamRatio Therapeutics, Inc.Ratio Therapeutics, Inc.Department of Radiology & Nuclear Medicine, Erasmus MC University Medical Centre RotterdamAbstract Background Fibroblast activation protein (FAP) is an attractive target for cancer theranostics. Although FAP-targeted nuclear imaging demonstrated promising clinical results, only sub-optimal results are reported for targeted radionuclide therapy (TRT). Preclinical research is crucial in selecting promising FAP-targeted radiopharmaceuticals and for obtaining an increased understanding of factors essential for FAP-TRT improvement. FAP is mainly expressed by cancer-associated fibroblasts in the tumor stroma and less on cancer cells themselves. Therefore, other (complex) factors impact FAP-TRT efficacy compared to currently clinically applied TRT strategies. For accurate evaluation of these aspects, selection of a representative preclinical model is important. Currently mainly human cancer cell lines transduced to (over)express FAP are applied, lacking clinical representation. It is unclear how these and more physiological FAP-expressing models compare to each other, and whether/how the model influences the study outcome. We aimed to address this by comparing FAP tracer behavior in FAP-transduced HT1080-huFAP and HEK293-huFAP cells, and endogenous FAP-expressing U-87 MG cancer cells and PS-1 pancreatic stellate cells. [111In]In-FAPI-46 and a fluorescent FAP-targeted tracer (RTX-1370S) were used to compare tracer binding/uptake and localization in vitro and ex vivo. Additionally, FAP expression was determined with RT-qPCR and anti-FAP IHC. Results Although FAP expression was highest in HEK293-huFAP cells and cell line derived xenografts, this did not result in the highest tracer uptake. [111In]In-FAPI-46 uptake was highest in HT1080-huFAP, closely followed by HEK293-huFAP, and a 6-10-fold lower uptake for U-87 MG and PS-1 cells. However, ex vivo U-87 MG xenografts only showed a 2-fold lower binding compared to HT1080-huFAP and HEK293-huFAP xenografts, mainly because the cell line attracts murine fibroblasts as demonstrated in our RT-qPCR and IHC studies. Conclusions The interaction between FAP and FAP-targeted tracers differs between models, indicating the need for appropriate model selection and that comparing results across studies using different models is difficult.https://doi.org/10.1186/s13550-024-01191-6Cancer-associated fibroblast (CAF)Fibroblast activation protein (FAP)Targeted radionuclide therapy (TRT)Radionuclide theranosticsPreclinical models
spellingShingle Circe D. van der Heide
Joana D. Campeiro
Eline A. M. Ruigrok
Lilian van den Brink
Shashikanth Ponnala
Shawn M. Hillier
Simone U. Dalm
In vitro and ex vivo evaluation of preclinical models for FAP-targeted theranostics: differences and relevance for radiotracer evaluation
EJNMMI Research
Cancer-associated fibroblast (CAF)
Fibroblast activation protein (FAP)
Targeted radionuclide therapy (TRT)
Radionuclide theranostics
Preclinical models
title In vitro and ex vivo evaluation of preclinical models for FAP-targeted theranostics: differences and relevance for radiotracer evaluation
title_full In vitro and ex vivo evaluation of preclinical models for FAP-targeted theranostics: differences and relevance for radiotracer evaluation
title_fullStr In vitro and ex vivo evaluation of preclinical models for FAP-targeted theranostics: differences and relevance for radiotracer evaluation
title_full_unstemmed In vitro and ex vivo evaluation of preclinical models for FAP-targeted theranostics: differences and relevance for radiotracer evaluation
title_short In vitro and ex vivo evaluation of preclinical models for FAP-targeted theranostics: differences and relevance for radiotracer evaluation
title_sort in vitro and ex vivo evaluation of preclinical models for fap targeted theranostics differences and relevance for radiotracer evaluation
topic Cancer-associated fibroblast (CAF)
Fibroblast activation protein (FAP)
Targeted radionuclide therapy (TRT)
Radionuclide theranostics
Preclinical models
url https://doi.org/10.1186/s13550-024-01191-6
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