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Preclinical SPECT Imaging of Choroidal Neovascularization in Mice Using Integrin-Binding [99mTc]IDA-D-[c(RGDfK)]2

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Abstract

Purpose

Integrin ɑvβ3, an adhesion molecule overexpressed in neovascular endothelial cells, is involved in ocular angiogenesis. Integrin ɑvβ3-binding arginine–glycine–aspartic acid (RGD) peptide has been used to target and visualize new vessels. We explored the use of integrin ɑvβ3-targeted RGD peptide ([99mTc]IDA-D-[c(RGDfK)]2) for in vivo molecular imaging of choroidal neovascularization (CNV).

Procedures

To induce CNV in animals, the right eyes of C57BL/6 mice were treated with retinal argon laser photocoagulation. CNV formation was confirmed on immunohistopathological examination of retinal and choroidal tissues. To explore the association of integrin with angiogenesis, integrin mRNA expression in the retinal and choroidal tissues was measured using real-time reverse transcriptase-polymerase chain reaction. For in vivo imaging, mice were intravenously injected with [99mTc]IDA-D-[c(RGDfK)]2 and single-photon emission computed tomography (SPECT) images of [99mTc]IDA-D-[c(RGDfK)]2 were obtained before laser induction (baseline) and at 1, 3, 7, and 14 days post-induction. CNV-induced regional alterations were measured using radiotracer uptake count.

Results

Immunohistopathological examination revealed that CNV lesions showed intense fluorescein isothiocyanate (FITC)-D-[c(RGDfK)]2 immunofluorescence, in contrast to the normal retina and choroid. Retinal integrin mRNA expression peaked at day 1 following CNV induction. On SPECT images using [99mTc]IDA-D-[c(RGDfK)]2, the radio-uptake count in eyes with CNV was significantly higher than in normal controls on days 1–7 (all p < 0.05), with a peak at day 3 representing the highest angiogenic activity. Our preclinical data demonstrated that [99mTc]IDA-D-[c(RGDfK)]2 can detect CNV and its associated angiogenesis in an animal model of CNV.

Conclusions

SPECT imaging using an integrin ɑvβ3-targeted RGD peptide radiotracer may be a useful tool for in vivo functional molecular imaging of CNV.

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Funding

This study was supported by the Korean Research Foundation [grant numbers NRF-2014R1A2A2A01007980, 2016R1D1A1B03934724, and 2018M2A2B3A02071842] and the Korea Health Technology R&D Project through the Korea Health Industry Development Institute, funded by the Ministry of Health & Welfare, Republic of Korea [HI14C1072].

Author information

Author notes

  1. Seong Joon Ahn, Ho-Young Lee and Hye Kyoung Hong contributed equally to this work.

Authors and Affiliations

  1. Department of Ophthalmology, Seoul National University Bundang Hospital, Seoul National University College of Medicine, 300 Gumidong, Bundanggu, Seongnam, 13620, Republic of Korea

    Seong Joon Ahn, Hye Kyoung Hong, Ji Hyun Park, Kyu Hyung Park & Se Joon Woo

  2. Department of Ophthalmology, Hanyang University Hospital, 222-1 Wangsimni-ro, Seongdong-gu, Seoul, 04763, Republic of Korea

    Seong Joon Ahn

  3. Department of Nuclear Medicine, Seoul National University Bundang Hospital, Seoul National University College of Medicine, 300 Gumidong, Bundanggu, Seongnam, 13620, Republic of Korea

    Ho-Young Lee, Jae Ho Jung, Sang Eun Kim & Byung Chul Lee

  4. Center for Nanomolecular Imaging and Innovative Drug Development, Advanced Institutes of Convergence Technology, Suwon, 16229, Republic of Korea

    Sang Eun Kim & Byung Chul Lee

  5. Department of Transdisciplinary Studies, Graduate School of Convergence Science and Technology, Seoul National University, Suwon, 16229, Republic of Korea

    Sang Eun Kim

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  1. Seong Joon Ahn
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Correspondence to Se Joon Woo or Byung Chul Lee.

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Ahn, S.J., Lee, HY., Hong, H.K. et al. Preclinical SPECT Imaging of Choroidal Neovascularization in Mice Using Integrin-Binding [99mTc]IDA-D-[c(RGDfK)]2. Mol Imaging Biol 21, 644–653 (2019). https://doi.org/10.1007/s11307-018-1294-8

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  • DOI: https://doi.org/10.1007/s11307-018-1294-8

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