Pretargeting of internalizing trastuzumab and cetuximab with a 18F-tetrazine tracer in xenograft models

Outi Keinänen, Kimberly E Fung, Jacob Pourat, Vilma Jallinoja, Delphine Vivier, NagaVara Kishore Pillarsetty, Anu Airaksinen, Jason S Lewis, Brian M Zeglis, Mirkka Sarparanta

Research output: Contribution to journalArticleScientificpeer-review

Abstract

Pretargeting-based approaches are being investigated for radioimmunoimaging and therapy applications to reduce the effective radiation burden to the patient. To date, only a few studies have used short-lived radioisotopes for pretargeting of antibodies, and such examples with internalizing antibodies are even rarer. Herein, we have investigated pretargeting methodology using inverse electron-demand Diels-Alder (IEDDA) for tracing two clinically relevant, internalizing monoclonal antibodies, cetuximab and trastuzumab.

Bioorthogonal reaction between tetrazine and trans-cyclooctene (TCO) was used for tracing cetuximab and trastuzumab in vivo with a fluorine-18 (t (A 1/2) = 109.8 min) labelled tracer. TCO-cetuximab or TCO-trastuzumab was administered 24, 48, or 72 h prior to the injection of tracer to A431 or BT-474 tumour-bearing mice, respectively. With cetuximab, the highest tumour-to-blood ratios were achieved when the lag time between antibody and tracer injections was 72 h. With trastuzumab, no difference was observed between different lag times. For both antibodies, the tumour could be clearly visualized in the PET images with the highest tumour uptake of 3.7 +/- 0.1%ID/g for cetuximab and 1.5 +/- 0.1%ID/g for trastuzumab as quantified by ex vivo biodistribution. In vivo IEDDA reaction was observed in the blood for both antibodies, but with trastuzumab, this was to a much lower degree than with cetuximab.

We could successfully visualize the tumours by using cetuximab and trastuzumab in pretargeted PET imaging despite the challenging circumstances where the antibody is internalized and there is still some unbound antibody circulating in the blood flow. This clearly demonstrates the potential of a pretargeted approach for targeting internalizing antigens and warrants development of pharmacokinetic optimization of the biorthogonal reactants to this end.
Original languageEnglish
Article number95
JournalEJNMMI Research
Volume2017
Issue number7
Number of pages12
ISSN2191-219X
DOIs
Publication statusPublished - 2 Dec 2017
MoE publication typeA1 Journal article-refereed

Fields of Science

  • 116 Chemical sciences
  • positron emission tomography (PET)
  • radiolabeling
  • Bioorthogonal chemistry
  • pretargeting
  • monoclonal antibody
  • internalization

Cite this

Keinänen, Outi ; Fung, Kimberly E ; Pourat, Jacob ; Jallinoja, Vilma ; Vivier, Delphine ; Pillarsetty, NagaVara Kishore ; Airaksinen, Anu ; Lewis, Jason S ; Zeglis, Brian M ; Sarparanta, Mirkka . / Pretargeting of internalizing trastuzumab and cetuximab with a 18F-tetrazine tracer in xenograft models. In: EJNMMI Research. 2017 ; Vol. 2017, No. 7.
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title = "Pretargeting of internalizing trastuzumab and cetuximab with a 18F-tetrazine tracer in xenograft models",
abstract = "Pretargeting-based approaches are being investigated for radioimmunoimaging and therapy applications to reduce the effective radiation burden to the patient. To date, only a few studies have used short-lived radioisotopes for pretargeting of antibodies, and such examples with internalizing antibodies are even rarer. Herein, we have investigated pretargeting methodology using inverse electron-demand Diels-Alder (IEDDA) for tracing two clinically relevant, internalizing monoclonal antibodies, cetuximab and trastuzumab.Bioorthogonal reaction between tetrazine and trans-cyclooctene (TCO) was used for tracing cetuximab and trastuzumab in vivo with a fluorine-18 (t (A 1/2) = 109.8 min) labelled tracer. TCO-cetuximab or TCO-trastuzumab was administered 24, 48, or 72 h prior to the injection of tracer to A431 or BT-474 tumour-bearing mice, respectively. With cetuximab, the highest tumour-to-blood ratios were achieved when the lag time between antibody and tracer injections was 72 h. With trastuzumab, no difference was observed between different lag times. For both antibodies, the tumour could be clearly visualized in the PET images with the highest tumour uptake of 3.7 +/- 0.1{\%}ID/g for cetuximab and 1.5 +/- 0.1{\%}ID/g for trastuzumab as quantified by ex vivo biodistribution. In vivo IEDDA reaction was observed in the blood for both antibodies, but with trastuzumab, this was to a much lower degree than with cetuximab.We could successfully visualize the tumours by using cetuximab and trastuzumab in pretargeted PET imaging despite the challenging circumstances where the antibody is internalized and there is still some unbound antibody circulating in the blood flow. This clearly demonstrates the potential of a pretargeted approach for targeting internalizing antigens and warrants development of pharmacokinetic optimization of the biorthogonal reactants to this end.",
keywords = "116 Chemical sciences, positron emission tomography (PET), radiolabeling, Bioorthogonal chemistry, pretargeting, monoclonal antibody, internalization",
author = "Outi Kein{\"a}nen and Fung, {Kimberly E} and Jacob Pourat and Vilma Jallinoja and Delphine Vivier and Pillarsetty, {NagaVara Kishore} and Anu Airaksinen and Lewis, {Jason S} and Zeglis, {Brian M} and Mirkka Sarparanta",
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Keinänen, O, Fung, KE, Pourat, J, Jallinoja, V, Vivier, D, Pillarsetty, NK, Airaksinen, A, Lewis, JS, Zeglis, BM & Sarparanta, M 2017, 'Pretargeting of internalizing trastuzumab and cetuximab with a 18F-tetrazine tracer in xenograft models', EJNMMI Research, vol. 2017, no. 7, 95. https://doi.org/10.1186/s13550-017-0344-6

Pretargeting of internalizing trastuzumab and cetuximab with a 18F-tetrazine tracer in xenograft models. / Keinänen, Outi; Fung, Kimberly E; Pourat, Jacob; Jallinoja, Vilma; Vivier, Delphine; Pillarsetty, NagaVara Kishore; Airaksinen, Anu; Lewis, Jason S; Zeglis, Brian M; Sarparanta, Mirkka .

In: EJNMMI Research, Vol. 2017, No. 7, 95, 02.12.2017.

Research output: Contribution to journalArticleScientificpeer-review

TY - JOUR

T1 - Pretargeting of internalizing trastuzumab and cetuximab with a 18F-tetrazine tracer in xenograft models

AU - Keinänen, Outi

AU - Fung, Kimberly E

AU - Pourat, Jacob

AU - Jallinoja, Vilma

AU - Vivier, Delphine

AU - Pillarsetty, NagaVara Kishore

AU - Airaksinen, Anu

AU - Lewis, Jason S

AU - Zeglis, Brian M

AU - Sarparanta, Mirkka

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N2 - Pretargeting-based approaches are being investigated for radioimmunoimaging and therapy applications to reduce the effective radiation burden to the patient. To date, only a few studies have used short-lived radioisotopes for pretargeting of antibodies, and such examples with internalizing antibodies are even rarer. Herein, we have investigated pretargeting methodology using inverse electron-demand Diels-Alder (IEDDA) for tracing two clinically relevant, internalizing monoclonal antibodies, cetuximab and trastuzumab.Bioorthogonal reaction between tetrazine and trans-cyclooctene (TCO) was used for tracing cetuximab and trastuzumab in vivo with a fluorine-18 (t (A 1/2) = 109.8 min) labelled tracer. TCO-cetuximab or TCO-trastuzumab was administered 24, 48, or 72 h prior to the injection of tracer to A431 or BT-474 tumour-bearing mice, respectively. With cetuximab, the highest tumour-to-blood ratios were achieved when the lag time between antibody and tracer injections was 72 h. With trastuzumab, no difference was observed between different lag times. For both antibodies, the tumour could be clearly visualized in the PET images with the highest tumour uptake of 3.7 +/- 0.1%ID/g for cetuximab and 1.5 +/- 0.1%ID/g for trastuzumab as quantified by ex vivo biodistribution. In vivo IEDDA reaction was observed in the blood for both antibodies, but with trastuzumab, this was to a much lower degree than with cetuximab.We could successfully visualize the tumours by using cetuximab and trastuzumab in pretargeted PET imaging despite the challenging circumstances where the antibody is internalized and there is still some unbound antibody circulating in the blood flow. This clearly demonstrates the potential of a pretargeted approach for targeting internalizing antigens and warrants development of pharmacokinetic optimization of the biorthogonal reactants to this end.

AB - Pretargeting-based approaches are being investigated for radioimmunoimaging and therapy applications to reduce the effective radiation burden to the patient. To date, only a few studies have used short-lived radioisotopes for pretargeting of antibodies, and such examples with internalizing antibodies are even rarer. Herein, we have investigated pretargeting methodology using inverse electron-demand Diels-Alder (IEDDA) for tracing two clinically relevant, internalizing monoclonal antibodies, cetuximab and trastuzumab.Bioorthogonal reaction between tetrazine and trans-cyclooctene (TCO) was used for tracing cetuximab and trastuzumab in vivo with a fluorine-18 (t (A 1/2) = 109.8 min) labelled tracer. TCO-cetuximab or TCO-trastuzumab was administered 24, 48, or 72 h prior to the injection of tracer to A431 or BT-474 tumour-bearing mice, respectively. With cetuximab, the highest tumour-to-blood ratios were achieved when the lag time between antibody and tracer injections was 72 h. With trastuzumab, no difference was observed between different lag times. For both antibodies, the tumour could be clearly visualized in the PET images with the highest tumour uptake of 3.7 +/- 0.1%ID/g for cetuximab and 1.5 +/- 0.1%ID/g for trastuzumab as quantified by ex vivo biodistribution. In vivo IEDDA reaction was observed in the blood for both antibodies, but with trastuzumab, this was to a much lower degree than with cetuximab.We could successfully visualize the tumours by using cetuximab and trastuzumab in pretargeted PET imaging despite the challenging circumstances where the antibody is internalized and there is still some unbound antibody circulating in the blood flow. This clearly demonstrates the potential of a pretargeted approach for targeting internalizing antigens and warrants development of pharmacokinetic optimization of the biorthogonal reactants to this end.

KW - 116 Chemical sciences

KW - positron emission tomography (PET)

KW - radiolabeling

KW - Bioorthogonal chemistry

KW - pretargeting

KW - monoclonal antibody

KW - internalization

U2 - 10.1186/s13550-017-0344-6

DO - 10.1186/s13550-017-0344-6

M3 - Article

VL - 2017

JO - EJNMMI Research

JF - EJNMMI Research

SN - 2191-219X

IS - 7

M1 - 95

ER -