Molecular design of HER3-targeting affibody molecules: Influence of chelator and presence of HEHEHE-Tag on biodistribution of <sup>68</sup> Ga-labeled tracers

Abstract

Affibody-based imaging of HER3 is a promising approach for patient stratification. We investigated the influence of a hydrophilic HEHEHE-tag ((HE) ₃ -tag) and two different gallium-68/chelator-complexes on the biodistribution of Z ₀₈₆₉₈ with the aim to improve the tracer for PET imaging. Affibody molecules (HE) ₃ -Z ₀₈₆₉₈ -X and Z ₀₈₆₉₈ -X (X = NOTA, NODAGA) were produced and labeled with gallium-68. Binding specificity and cellular processing were studied in HER3-expressing human cancer cell lines BxPC-3 and DU145. Biodistribution was studied 3 h p.i. in Balb/c nu/nu mice bearing BxPC-3 xenografts. Mice were imaged 3 h p.i. using microPET/CT. Conjugates were stably labeled with gallium-68 and bound specifically to HER3 in vitro and in vivo. Association to cells was rapid but internalization was slow. Uptake in tissues, including tumors, was lower for (HE) ₃ -Z ₀₈₆₉₈ -X than for non-tagged variants. The neutral [ ⁶⁸ Ga]Ga-NODAGA complex reduced the hepatic uptake of Z ₀₈₆₉₈ compared to positively charged [ ⁶⁸ Ga]Ga-NOTA-conjugated variants. The influence of the chelator was more pronounced in variants without (HE) _3- tag. In conclusion, hydrophilic (HE) ₃ -tag and neutral charge of the [ ⁶⁸ Ga]Ga-NODAGA complex promoted blood clearance and lowered hepatic uptake of Z ₀₈₆₉₈ . [ ⁶⁸ Ga]Ga-(HE) ₃ -Z ₀₈₆₉₈ -NODAGA was considered most promising, providing the lowest blood and hepatic uptake and the best imaging contrast among the tested variants.

Original language	English
Article number	1080
Journal	International Journal of Molecular Sciences
Volume	20
Issue number	5
DOIs	https://doi.org/10.3390/ijms20051080
Publication status	Published - Mar 2019
Externally published	Yes

Keywords

Affibody
Gallium-68
HER3
Molecular imaging
NODAGA
NOTA
PET

ASJC Scopus subject areas

Catalysis
Molecular Biology
Spectroscopy
Computer Science Applications
Physical and Theoretical Chemistry
Organic Chemistry
Inorganic Chemistry

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Leitao, C. D., Rinne, S. S., Mitran, B., Vorobyeva, A., Andersson, K. G., Tolmachev, V., Ståhl, S., Löfblom, J., & Orlova, A. (2019). Molecular design of HER3-targeting affibody molecules: Influence of chelator and presence of HEHEHE-Tag on biodistribution of ⁶⁸ Ga-labeled tracers. International Journal of Molecular Sciences, 20(5), [1080]. https://doi.org/10.3390/ijms20051080

Molecular design of HER3-targeting affibody molecules : Influence of chelator and presence of HEHEHE-Tag on biodistribution of ⁶⁸ Ga-labeled tracers. / Leitao, Charles Dahlsson; Rinne, Sara S.; Mitran, Bogdan; Vorobyeva, Anzhelika; Andersson, Ken G.; Tolmachev, Vladimir; Ståhl, Stefan; Löfblom, John; Orlova, Anna.

In: International Journal of Molecular Sciences, Vol. 20, No. 5, 1080, 03.2019.

Research output: Contribution to journal › Article › peer-review

Leitao, CD, Rinne, SS, Mitran, B, Vorobyeva, A, Andersson, KG, Tolmachev, V, Ståhl, S, Löfblom, J & Orlova, A 2019, 'Molecular design of HER3-targeting affibody molecules: Influence of chelator and presence of HEHEHE-Tag on biodistribution of ⁶⁸ Ga-labeled tracers', International Journal of Molecular Sciences, vol. 20, no. 5, 1080. https://doi.org/10.3390/ijms20051080

Leitao CD, Rinne SS, Mitran B, Vorobyeva A, Andersson KG, Tolmachev V et al. Molecular design of HER3-targeting affibody molecules: Influence of chelator and presence of HEHEHE-Tag on biodistribution of ⁶⁸ Ga-labeled tracers. International Journal of Molecular Sciences. 2019 Mar;20(5). 1080. https://doi.org/10.3390/ijms20051080

Leitao, Charles Dahlsson ; Rinne, Sara S. ; Mitran, Bogdan ; Vorobyeva, Anzhelika ; Andersson, Ken G. ; Tolmachev, Vladimir ; Ståhl, Stefan ; Löfblom, John ; Orlova, Anna. / Molecular design of HER3-targeting affibody molecules : Influence of chelator and presence of HEHEHE-Tag on biodistribution of ⁶⁸ Ga-labeled tracers. In: International Journal of Molecular Sciences. 2019 ; Vol. 20, No. 5.

@article{82c697c798cd447eb32ee05af2dc269e,

title = "Molecular design of HER3-targeting affibody molecules: Influence of chelator and presence of HEHEHE-Tag on biodistribution of 68 Ga-labeled tracers",

abstract = " Affibody-based imaging of HER3 is a promising approach for patient stratification. We investigated the influence of a hydrophilic HEHEHE-tag ((HE) 3 -tag) and two different gallium-68/chelator-complexes on the biodistribution of Z 08698 with the aim to improve the tracer for PET imaging. Affibody molecules (HE) 3 -Z 08698 -X and Z 08698 -X (X = NOTA, NODAGA) were produced and labeled with gallium-68. Binding specificity and cellular processing were studied in HER3-expressing human cancer cell lines BxPC-3 and DU145. Biodistribution was studied 3 h p.i. in Balb/c nu/nu mice bearing BxPC-3 xenografts. Mice were imaged 3 h p.i. using microPET/CT. Conjugates were stably labeled with gallium-68 and bound specifically to HER3 in vitro and in vivo. Association to cells was rapid but internalization was slow. Uptake in tissues, including tumors, was lower for (HE) 3 -Z 08698 -X than for non-tagged variants. The neutral [ 68 Ga]Ga-NODAGA complex reduced the hepatic uptake of Z 08698 compared to positively charged [ 68 Ga]Ga-NOTA-conjugated variants. The influence of the chelator was more pronounced in variants without (HE) 3- tag. In conclusion, hydrophilic (HE) 3 -tag and neutral charge of the [ 68 Ga]Ga-NODAGA complex promoted blood clearance and lowered hepatic uptake of Z 08698 . [ 68 Ga]Ga-(HE) 3 -Z 08698 -NODAGA was considered most promising, providing the lowest blood and hepatic uptake and the best imaging contrast among the tested variants. ",

keywords = "Affibody, Gallium-68, HER3, Molecular imaging, NODAGA, NOTA, PET",

author = "Leitao, {Charles Dahlsson} and Rinne, {Sara S.} and Bogdan Mitran and Anzhelika Vorobyeva and Andersson, {Ken G.} and Vladimir Tolmachev and Stefan St{\aa}hl and John L{\"o}fblom and Anna Orlova",

note = "Funding Information: Funding: This study was supported by the Swedish Cancer Society (grants CAN2014/474 and CAN 2017/425 (A.O.), CAN2015/350 and CAN 2018/436 (V.T.), CAN2017/649 (J.L.) and CAN2016/463 (S.S.)), the Swedish Research Council (grants 2015-02509 (A.O.), 2015-02353 (V.T.) and 2012-05236 (S.S.)), the Swedish Agency for Innovation VINNOVA (grant 2016/04060 (A.O.)) and the Wallenberg Center for Protein Technology (S.S. and J.L.) which are acknowledged for financial support. Funding Information: Acknowledgments: The molecular imaging work in this publication was supported by the Wallenberg infrastructure for PET-MRI (WIPPET) at SciLifeLab Pilot Facility for Preclinical PET-MRI, a Swedish nationally available imaging platform at Uppsala University, Sweden, financed by Knut and Alice Wallenberg Foundation (SPECT/CT). Publisher Copyright: {\textcopyright} 2019 by the authors. Licensee MDPI, Basel, Switzerland. Copyright: Copyright 2019 Elsevier B.V., All rights reserved.",

year = "2019",

month = mar,

doi = "10.3390/ijms20051080",

language = "English",

volume = "20",

journal = "International Journal of Molecular Sciences",

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T1 - Molecular design of HER3-targeting affibody molecules

T2 - Influence of chelator and presence of HEHEHE-Tag on biodistribution of 68 Ga-labeled tracers

AU - Leitao, Charles Dahlsson

AU - Rinne, Sara S.

AU - Mitran, Bogdan

AU - Vorobyeva, Anzhelika

AU - Andersson, Ken G.

AU - Tolmachev, Vladimir

AU - Ståhl, Stefan

AU - Löfblom, John

AU - Orlova, Anna

N1 - Funding Information: Funding: This study was supported by the Swedish Cancer Society (grants CAN2014/474 and CAN 2017/425 (A.O.), CAN2015/350 and CAN 2018/436 (V.T.), CAN2017/649 (J.L.) and CAN2016/463 (S.S.)), the Swedish Research Council (grants 2015-02509 (A.O.), 2015-02353 (V.T.) and 2012-05236 (S.S.)), the Swedish Agency for Innovation VINNOVA (grant 2016/04060 (A.O.)) and the Wallenberg Center for Protein Technology (S.S. and J.L.) which are acknowledged for financial support. Funding Information: Acknowledgments: The molecular imaging work in this publication was supported by the Wallenberg infrastructure for PET-MRI (WIPPET) at SciLifeLab Pilot Facility for Preclinical PET-MRI, a Swedish nationally available imaging platform at Uppsala University, Sweden, financed by Knut and Alice Wallenberg Foundation (SPECT/CT). Publisher Copyright: © 2019 by the authors. Licensee MDPI, Basel, Switzerland. Copyright: Copyright 2019 Elsevier B.V., All rights reserved.

PY - 2019/3

Y1 - 2019/3

N2 - Affibody-based imaging of HER3 is a promising approach for patient stratification. We investigated the influence of a hydrophilic HEHEHE-tag ((HE) 3 -tag) and two different gallium-68/chelator-complexes on the biodistribution of Z 08698 with the aim to improve the tracer for PET imaging. Affibody molecules (HE) 3 -Z 08698 -X and Z 08698 -X (X = NOTA, NODAGA) were produced and labeled with gallium-68. Binding specificity and cellular processing were studied in HER3-expressing human cancer cell lines BxPC-3 and DU145. Biodistribution was studied 3 h p.i. in Balb/c nu/nu mice bearing BxPC-3 xenografts. Mice were imaged 3 h p.i. using microPET/CT. Conjugates were stably labeled with gallium-68 and bound specifically to HER3 in vitro and in vivo. Association to cells was rapid but internalization was slow. Uptake in tissues, including tumors, was lower for (HE) 3 -Z 08698 -X than for non-tagged variants. The neutral [ 68 Ga]Ga-NODAGA complex reduced the hepatic uptake of Z 08698 compared to positively charged [ 68 Ga]Ga-NOTA-conjugated variants. The influence of the chelator was more pronounced in variants without (HE) 3- tag. In conclusion, hydrophilic (HE) 3 -tag and neutral charge of the [ 68 Ga]Ga-NODAGA complex promoted blood clearance and lowered hepatic uptake of Z 08698 . [ 68 Ga]Ga-(HE) 3 -Z 08698 -NODAGA was considered most promising, providing the lowest blood and hepatic uptake and the best imaging contrast among the tested variants.

AB - Affibody-based imaging of HER3 is a promising approach for patient stratification. We investigated the influence of a hydrophilic HEHEHE-tag ((HE) 3 -tag) and two different gallium-68/chelator-complexes on the biodistribution of Z 08698 with the aim to improve the tracer for PET imaging. Affibody molecules (HE) 3 -Z 08698 -X and Z 08698 -X (X = NOTA, NODAGA) were produced and labeled with gallium-68. Binding specificity and cellular processing were studied in HER3-expressing human cancer cell lines BxPC-3 and DU145. Biodistribution was studied 3 h p.i. in Balb/c nu/nu mice bearing BxPC-3 xenografts. Mice were imaged 3 h p.i. using microPET/CT. Conjugates were stably labeled with gallium-68 and bound specifically to HER3 in vitro and in vivo. Association to cells was rapid but internalization was slow. Uptake in tissues, including tumors, was lower for (HE) 3 -Z 08698 -X than for non-tagged variants. The neutral [ 68 Ga]Ga-NODAGA complex reduced the hepatic uptake of Z 08698 compared to positively charged [ 68 Ga]Ga-NOTA-conjugated variants. The influence of the chelator was more pronounced in variants without (HE) 3- tag. In conclusion, hydrophilic (HE) 3 -tag and neutral charge of the [ 68 Ga]Ga-NODAGA complex promoted blood clearance and lowered hepatic uptake of Z 08698 . [ 68 Ga]Ga-(HE) 3 -Z 08698 -NODAGA was considered most promising, providing the lowest blood and hepatic uptake and the best imaging contrast among the tested variants.

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KW - Molecular imaging

KW - NODAGA

KW - NOTA

KW - PET

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