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Preparation and bioevaluation of 99mTc-HYNIC-annexin B1 as a novel radioligand for apoptosis imaging

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Abstract

Annexin B1, a novel Ca2+-dependent PS-binding protein, has been shown to have a high affinity for PS exposed on the surface of apoptotic cells. To develop and bioevaluate an annexin B1 based PS-targeting radiotracer, annexin B1 was radiolabeled with 99mTc using HYNIC as a bifunctional chelator. Binding assays with activated platelets and apoptotic SP2/0 cells were carried out to evaluate the in vitro biological activity of 99mTc-HYNIC-annexin B1. Biodistribution of this radioligand was studied in normal mice. Dexamethasone-induced murine thymus apoptosis and fas-mediated murine liver apoptosis models were used to investigate the ability of radiolabeled annexin B1 to detect apoptosis in vivo. The labeling procedure yielded a compound with up to 98% radiochemical purity and good in vitro stability. The in vitro binding assays indicated that 99mTc-HYNIC-annexin B1 retain its PS-binding activity. Biodistribution of the compound in mice showed that 99mTc-HYNIC-annexin B1 is rapidly cleared from the blood and predominantly accumulates in the kidney. The marked increase in dexamethasone-treated murine thymus uptake and fas-mediated murine liver uptake correlated with histologic evidence of apoptosis. These data suggested that 99mTc-HYNIC-annexin B1 retain its in vitro and in vivo biological activities. This radiotracer may therefore be useful as a novel radioligand for the noninvasive detecting of PS externalization associated with apoptosis.

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Abbreviations

FITC:

Fluorescein isothiocyanate

FPLC:

Fast protein liquid chromatography

GST:

Glutathione-S-transferase

HYNIC:

Hydrazinonicotinamide

HE:

Hematoxylin–Eosin

%ID/g:

Percent of injected dose per gram

ITLC-SG:

Instant thin-layer chromatography on silica gel

LCM:

Laser confocal microscopy

MSR:

Molar substitution rate

NHS-HYNIC:

N-Hydroxysuccinimidyl-hydrazinonicotinamide

PI:

Propidium iodide

PS:

Phosphatidylserine

RCP:

Radiochemical purity

SDS-PAGE:

Sodium dodecyl sulfate-polyacrylamide gel electrophoresis

SPECT:

Single photon emission computerized tomography

TUNEL:

TdT-mediated dUTP nick-end-labeling

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Acknowledgements

The authors thank Mr. Liu Xing-feng and Ms. Luo Qiong for their technical help and assistance for this study. This work was financially supported by grants from the National Natural Science Foundation of China (30600155) and the Shanghai Rising-Star Program of China (06QA14040).

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Authors and Affiliations

  1. Department of Nuclear Medicine, Shanghai Sixth People’s Hospital, Shanghai Jiao Tong University, 600 Yishan Road, Shanghai, 200233, China

    Quan-Yong Luo, Han-Kui Lu & Rui-Sen Zhu

  2. Department of Medical Genetics, Second Military Medical University, Shanghai, 200433, China

    Fang Wang, Yi Zhang & Shu-Han Sun

  3. Radiopharmaceutical Lab, Shanghai GMS Pharmaceutical Co., Ltd., Shanghai, 201108, China

    Zhi-Yong Zhang

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  1. Quan-Yong Luo
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  2. Fang Wang
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Correspondence to Quan-Yong Luo.

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Luo, QY., Wang, F., Zhang, ZY. et al. Preparation and bioevaluation of 99mTc-HYNIC-annexin B1 as a novel radioligand for apoptosis imaging. Apoptosis 13, 600–608 (2008). https://doi.org/10.1007/s10495-008-0189-6

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