Assuntos
Anexinas/farmacocinética , Apoptose , Glaucoma/diagnóstico por imagem , Imagem Óptica/métodos , Células Ganglionares da Retina , Retinoscopia/métodos , Adulto , Anexinas/administração & dosagem , Anexinas/efeitos adversos , Feminino , Corantes Fluorescentes , Humanos , Masculino , Pessoa de Meia-Idade , Células Ganglionares da Retina/citologiaRESUMO
A novel annexin A5 derivative (cys-annexin A5) with a single cysteine residue at its C-terminal has been developed and successfully labeled in high labeling yield with (99m)Tc by a ligand exchange reaction. Like the 1st generation (99m)Tc-HYNIC-annexin A5, the novel (99m)Tc-cys-annexin A5 derivative shows in normal mice mainly renal and, to a lesser extent, hepatobiliary excretion. In rat models of hepatic apoptosis there was 283% increase in hepatic uptake of (99m)Tc-cys-annexin A5 as compared to normal mice. The results indicate that the novel (99m)Tc-cys-annexin A5 is a potential apoptosis imaging agent.
Assuntos
Anexinas/química , Apoptose , Compostos de Organotecnécio/química , Compostos Radiofarmacêuticos/química , Animais , Anexina A5/química , Anexinas/farmacocinética , Anexinas/toxicidade , Marcação In Situ das Extremidades Cortadas , Marcação por Isótopo , Fígado/metabolismo , Fígado/patologia , Masculino , Camundongos , Compostos de Organotecnécio/farmacocinética , Compostos de Organotecnécio/toxicidade , Compostos Radiofarmacêuticos/farmacocinética , Compostos Radiofarmacêuticos/toxicidade , Ratos , Distribuição Tecidual , Tomografia Computadorizada de Emissão de Fóton ÚnicoRESUMO
BACKGROUND: A novel dual-contrast molecular MRI technique to image both cardiomyocyte apoptosis and necrosis in vivo within 4 to 6 hours of ischemia is presented. The technique uses the annexin-based nanoparticle AnxCLIO-Cy5.5 (apoptosis) and simultaneous delayed-enhancement imaging with a novel gadolinium chelate, Gd-DTPA-NBD (necrosis). METHODS AND RESULTS: Mice with transient coronary ligation were injected intravenously at the onset of reperfusion with AnxCLIO-Cy5.5 (n=7) or the control probe Inact_CLIO-Cy5.5 (n=6). T2*-weighted MR images (9.4 T) were acquired within 4 to 6 hours of reperfusion. The contrast-to-noise ratio between injured and uninjured myocardium was measured. The mice were then injected with Gd-DTPA-NBD, and delayed-enhancement imaging was performed within 10 to 30 minutes. Uptake of AnxCLIO-Cy5.5 was most prominent in the midmyocardium and was significantly greater than that of Inact_CLIO-Cy5.5 (contrast-to-noise ratio, 8.82+/-1.5 versus 3.78+/-1.1; P<0.05). Only 21+/-3% of the myocardium with accumulation of AnxCLIO-Cy5.5 showed delayed-enhancement of Gd-DTPA-NBD. Wall thickening was significantly reduced in segments with delayed enhancement and/or transmural accumulation of AnxCLIO-Cy5.5 (P<0.001). Fluorescence microscopy of AnxCLIO-Cy5.5 and immunohistochemistry of Gd-DTPA-NBD confirmed the presence of large numbers of apoptotic but potentially viable cardiomyocytes (AnxCLIO-Cy5.5 positive, Gd-DTPA-NBD negative) in the midmyocardium. CONCLUSIONS: A novel technique to image cardiomyocyte apoptosis and necrosis in vivo within 4 to 6 hours of injury is presented and reveals large areas of apoptotic but viable myocardium in the midmyocardium. Strategies to salvage the numerous apoptotic but potentially viable cardiomyocytes in the midmyocardium in acute ischemia should be investigated.
Assuntos
4-Cloro-7-nitrobenzofurazano/análogos & derivados , Anexina A5/farmacocinética , Anexinas/farmacocinética , Apoptose , Carbocianinas/farmacocinética , Meios de Contraste/farmacocinética , Gadolínio DTPA/análogos & derivados , Imageamento por Ressonância Magnética/métodos , Infarto do Miocárdio/patologia , Miócitos Cardíacos/patologia , Nanopartículas , Necrose/diagnóstico , Compostos Organometálicos/farmacocinética , Ácido Pentético/análogos & derivados , 4-Cloro-7-nitrobenzofurazano/farmacocinética , Análise de Variância , Animais , Citometria de Fluxo , Gadolínio DTPA/farmacocinética , Processamento de Imagem Assistida por Computador , Marcação In Situ das Extremidades Cortadas , Camundongos , Camundongos Endogâmicos C57BL , Microscopia de Fluorescência , Nanotecnologia , Ácido Pentético/farmacocinética , Estatísticas não ParamétricasRESUMO
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 (99m)Tc 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 (99m)Tc-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 (99m)Tc-HYNIC-annexin B1 retain its PS-binding activity. Biodistribution of the compound in mice showed that (99m)Tc-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 (99m)Tc-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.
Assuntos
Anexinas/síntese química , Apoptose/fisiologia , Rim/citologia , Fígado/citologia , Compostos de Organotecnécio/síntese química , Fosfatidilserinas/metabolismo , Timo/citologia , Animais , Anexinas/farmacocinética , Apoptose/efeitos dos fármacos , Plaquetas/metabolismo , Dexametasona/farmacologia , Proteína Ligante Fas/farmacologia , Humanos , Ligantes , Camundongos , Compostos de Organotecnécio/farmacocinéticaRESUMO
To develop a radiopharmaceutical for apoptosis imaging, Annexin B1, a new Ca2+-dependent phosphatidylserine (PS)-binding protein, was directly radiolabeled with (99m)Tc. This procedure yields up to 96% of radiochemical purity and higher radiolabeling efficiency. The preparation has been found to be sufficiently stable in vitro. Binding assay with human activated platelets indicated that (99m)Tc-Annexin B1 retained its PS binding activity. Biodistribution in mice revealed that (99m)Tc-Annexin B1 rapidly cleared from the blood and predominantly accumulated in the kidney. The increase in hepatic uptake in anti-Fas antibody treated mice correlated to histologic evidence of fulminant hepatic apoptosis. These data suggest that (99m)Tc-Annexin B1 can be used as a novel radiotracer to detect apoptosis in vivo.
Assuntos
Anexinas/farmacocinética , Apoptose/fisiologia , Plaquetas/diagnóstico por imagem , Plaquetas/fisiologia , Compostos de Organotecnécio/farmacocinética , Animais , Anexinas/química , Células Cultivadas , Estabilidade de Medicamentos , Humanos , Marcação por Isótopo/métodos , Camundongos , Camundongos Endogâmicos BALB C , Especificidade de Órgãos , Compostos de Organotecnécio/química , Cintilografia , Compostos Radiofarmacêuticos/química , Compostos Radiofarmacêuticos/farmacocinética , Reprodutibilidade dos Testes , Distribuição TecidualRESUMO
State-of-the-art techniques have been used to measure key aspects of cardiovascular pathophysiology from the birth of radionuclide cardiovascular imaging. However, during the last 30 years, there have been few innovative imaging advances to further our understanding of the complex physiologic processes. Molecular imaging now offers an array of tools to develop advanced diagnostic approaches and therapies for patients with coronary artery disease and heart failure. For example, the enhanced understanding of the pathophysiology of atheroma makes it possible to identify vulnerable plaque based on its metabolic signature or the presence of excessive apoptosis. Because the metabolic and apoptotic signals are large, it is likely that even small lesions will be visible. Of the many approaches that are being developed, 2 tracers appear most likely to be tested in the near future: (1) [18F]-fluorodeoxyglucose, to determine macrophage metabolism; and (2) radiolabeled annexin, to measure apoptosis of the inflammatory cells. Using existing techniques such as perfusion imaging, appropriate patients can be selected for treatment with novel therapies, such as stem cell transplantation or vascular gene therapy. Using positron tomography in place of single photon imaging adds the capability for the measurement of absolute perfusion and perfusion reserve to the information on regional perfusion. Flow reserve detects global decreases in perfusion and refines the determination of lesion severity available from perfusion imaging.