Highly bright and stable NIR-BRET with blue-shifted coelenterazine derivatives for deep-tissue imaging of molecular events in vivo

Ryo Nishihara, Ramasamy Paulmurugan, Takahiro Nakajima, Eiji Yamamoto, Arutselvan Natarajan, Rayhaneh Afjei, Yuki Hiruta, Naoko Iwasawa, Shigeru Nishiyama, Daniel Citterio, Moritoshi Sato, Sung Bae Kim, Koji Suzuki

Research output: Contribution to journalArticle

Abstract

Background: Bioluminescence imaging (BLI) is one of the most widely used optical platforms in molecular imaging, but it suffers from severe tissue attenuation and autoluminescence in vivo. Methods: Here, we developed a novel BLI platform on the basis of bioluminescence resonance energy transfer (BRET) for achieving a ∼300 nm blue-to-near infrared shift of the emission (NIR-BRET) by synthesizing an array of 18 novel coelenterazine (CTZ) derivatives, named “Bottle Blue (BBlue)” and a unique iRFP-linked RLuc8.6-535SG fusion protein as a probe. Results: The best NIR-BRET was achieved by tuning the emission peaks of the CTZ derivatives to a Soret band of the iRFP. In mammalian cells, BBlue2.3, one of the CTZ derivatives, emits light that is ∼50-fold brighter than DBlueC when combined with RLuc8.6-535SG, which shows stable BL kinetics. When we used a caged version of BBLue2.3, it showed a BL half decay time of over 60 minutes while maintaining the higher signal sensitivity. This NIR BL is sufficiently brighter to be used for imaging live mammalian cells at single cell level, and also for imaging metastases in deep tissues in live mice without generating considerable autoluminescence. A single-chain probe developed based on this BLI platform allowed us to sensitively image ligand antagonist-specific activation of estrogen receptor in the NIR region. Conclusion: This unique optical platform provides the brightest NIR BLI template that can be used for imaging a diverse group of cellular events in living subjects including protein‒protein interactions and cancer metastasis.

Original languageEnglish
Pages (from-to)2646-2661
Number of pages16
JournalTheranostics
Volume9
Issue number9
DOIs
Publication statusPublished - 2019 Jan 1

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Molecular Imaging
Energy Transfer
Neoplasm Metastasis
Estrogen Receptors
Ligands
Light
coelenterazine
Neoplasms
Proteins

Keywords

  • Bioluminescence imaging
  • Bioluminescence resonance energy transfer (BRET)
  • Blue-to-near infrared shift
  • Coelenterazine derivatives
  • Metastasis

ASJC Scopus subject areas

  • Medicine (miscellaneous)
  • Pharmacology, Toxicology and Pharmaceutics (miscellaneous)

Cite this

Nishihara, R., Paulmurugan, R., Nakajima, T., Yamamoto, E., Natarajan, A., Afjei, R., ... Suzuki, K. (2019). Highly bright and stable NIR-BRET with blue-shifted coelenterazine derivatives for deep-tissue imaging of molecular events in vivo. Theranostics, 9(9), 2646-2661. https://doi.org/10.7150/thno.32219

Highly bright and stable NIR-BRET with blue-shifted coelenterazine derivatives for deep-tissue imaging of molecular events in vivo. / Nishihara, Ryo; Paulmurugan, Ramasamy; Nakajima, Takahiro; Yamamoto, Eiji; Natarajan, Arutselvan; Afjei, Rayhaneh; Hiruta, Yuki; Iwasawa, Naoko; Nishiyama, Shigeru; Citterio, Daniel; Sato, Moritoshi; Kim, Sung Bae; Suzuki, Koji.

In: Theranostics, Vol. 9, No. 9, 01.01.2019, p. 2646-2661.

Research output: Contribution to journalArticle

Nishihara, R, Paulmurugan, R, Nakajima, T, Yamamoto, E, Natarajan, A, Afjei, R, Hiruta, Y, Iwasawa, N, Nishiyama, S, Citterio, D, Sato, M, Kim, SB & Suzuki, K 2019, 'Highly bright and stable NIR-BRET with blue-shifted coelenterazine derivatives for deep-tissue imaging of molecular events in vivo', Theranostics, vol. 9, no. 9, pp. 2646-2661. https://doi.org/10.7150/thno.32219
Nishihara, Ryo ; Paulmurugan, Ramasamy ; Nakajima, Takahiro ; Yamamoto, Eiji ; Natarajan, Arutselvan ; Afjei, Rayhaneh ; Hiruta, Yuki ; Iwasawa, Naoko ; Nishiyama, Shigeru ; Citterio, Daniel ; Sato, Moritoshi ; Kim, Sung Bae ; Suzuki, Koji. / Highly bright and stable NIR-BRET with blue-shifted coelenterazine derivatives for deep-tissue imaging of molecular events in vivo. In: Theranostics. 2019 ; Vol. 9, No. 9. pp. 2646-2661.
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abstract = "Background: Bioluminescence imaging (BLI) is one of the most widely used optical platforms in molecular imaging, but it suffers from severe tissue attenuation and autoluminescence in vivo. Methods: Here, we developed a novel BLI platform on the basis of bioluminescence resonance energy transfer (BRET) for achieving a ∼300 nm blue-to-near infrared shift of the emission (NIR-BRET) by synthesizing an array of 18 novel coelenterazine (CTZ) derivatives, named “Bottle Blue (BBlue)” and a unique iRFP-linked RLuc8.6-535SG fusion protein as a probe. Results: The best NIR-BRET was achieved by tuning the emission peaks of the CTZ derivatives to a Soret band of the iRFP. In mammalian cells, BBlue2.3, one of the CTZ derivatives, emits light that is ∼50-fold brighter than DBlueC when combined with RLuc8.6-535SG, which shows stable BL kinetics. When we used a caged version of BBLue2.3, it showed a BL half decay time of over 60 minutes while maintaining the higher signal sensitivity. This NIR BL is sufficiently brighter to be used for imaging live mammalian cells at single cell level, and also for imaging metastases in deep tissues in live mice without generating considerable autoluminescence. A single-chain probe developed based on this BLI platform allowed us to sensitively image ligand antagonist-specific activation of estrogen receptor in the NIR region. Conclusion: This unique optical platform provides the brightest NIR BLI template that can be used for imaging a diverse group of cellular events in living subjects including protein‒protein interactions and cancer metastasis.",
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T1 - Highly bright and stable NIR-BRET with blue-shifted coelenterazine derivatives for deep-tissue imaging of molecular events in vivo

AU - Nishihara, Ryo

AU - Paulmurugan, Ramasamy

AU - Nakajima, Takahiro

AU - Yamamoto, Eiji

AU - Natarajan, Arutselvan

AU - Afjei, Rayhaneh

AU - Hiruta, Yuki

AU - Iwasawa, Naoko

AU - Nishiyama, Shigeru

AU - Citterio, Daniel

AU - Sato, Moritoshi

AU - Kim, Sung Bae

AU - Suzuki, Koji

PY - 2019/1/1

Y1 - 2019/1/1

N2 - Background: Bioluminescence imaging (BLI) is one of the most widely used optical platforms in molecular imaging, but it suffers from severe tissue attenuation and autoluminescence in vivo. Methods: Here, we developed a novel BLI platform on the basis of bioluminescence resonance energy transfer (BRET) for achieving a ∼300 nm blue-to-near infrared shift of the emission (NIR-BRET) by synthesizing an array of 18 novel coelenterazine (CTZ) derivatives, named “Bottle Blue (BBlue)” and a unique iRFP-linked RLuc8.6-535SG fusion protein as a probe. Results: The best NIR-BRET was achieved by tuning the emission peaks of the CTZ derivatives to a Soret band of the iRFP. In mammalian cells, BBlue2.3, one of the CTZ derivatives, emits light that is ∼50-fold brighter than DBlueC when combined with RLuc8.6-535SG, which shows stable BL kinetics. When we used a caged version of BBLue2.3, it showed a BL half decay time of over 60 minutes while maintaining the higher signal sensitivity. This NIR BL is sufficiently brighter to be used for imaging live mammalian cells at single cell level, and also for imaging metastases in deep tissues in live mice without generating considerable autoluminescence. A single-chain probe developed based on this BLI platform allowed us to sensitively image ligand antagonist-specific activation of estrogen receptor in the NIR region. Conclusion: This unique optical platform provides the brightest NIR BLI template that can be used for imaging a diverse group of cellular events in living subjects including protein‒protein interactions and cancer metastasis.

AB - Background: Bioluminescence imaging (BLI) is one of the most widely used optical platforms in molecular imaging, but it suffers from severe tissue attenuation and autoluminescence in vivo. Methods: Here, we developed a novel BLI platform on the basis of bioluminescence resonance energy transfer (BRET) for achieving a ∼300 nm blue-to-near infrared shift of the emission (NIR-BRET) by synthesizing an array of 18 novel coelenterazine (CTZ) derivatives, named “Bottle Blue (BBlue)” and a unique iRFP-linked RLuc8.6-535SG fusion protein as a probe. Results: The best NIR-BRET was achieved by tuning the emission peaks of the CTZ derivatives to a Soret band of the iRFP. In mammalian cells, BBlue2.3, one of the CTZ derivatives, emits light that is ∼50-fold brighter than DBlueC when combined with RLuc8.6-535SG, which shows stable BL kinetics. When we used a caged version of BBLue2.3, it showed a BL half decay time of over 60 minutes while maintaining the higher signal sensitivity. This NIR BL is sufficiently brighter to be used for imaging live mammalian cells at single cell level, and also for imaging metastases in deep tissues in live mice without generating considerable autoluminescence. A single-chain probe developed based on this BLI platform allowed us to sensitively image ligand antagonist-specific activation of estrogen receptor in the NIR region. Conclusion: This unique optical platform provides the brightest NIR BLI template that can be used for imaging a diverse group of cellular events in living subjects including protein‒protein interactions and cancer metastasis.

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KW - Bioluminescence resonance energy transfer (BRET)

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KW - Coelenterazine derivatives

KW - Metastasis

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