RESUMO
One of the main challenges of PET imaging with 89Zr-labeled monoclonal antibodies (mAbs) remains the long blood circulation of the radiolabeled mAbs, leading to high background signals, decreasing image quality. To overcome this limitation, here we report the use of a bioorthogonal linker cleavage approach (click-to-release chemistry) to selectively liberate [89Zr]Zr-DFO from trans-cyclooctene-functionalized trastuzumab (TCO-Tmab) in blood, following the administration of a tetrazine compound (trigger) in BT-474 tumor-bearing mice. Methods: We created a series of TCO-DFO constructs and evaluated their performance in [89Zr]Zr-DFO release from Tmab in vitro using different trigger compounds. The in vivo behavior of the best performing [89Zr]Zr-TCO-Tmab was studied in healthy mice first to determine the optimal dose of the trigger. To find the optimal time for the trigger administration, the rate of [89Zr]Zr-TCO-Tmab internalization was studied in BT-474 cancer cells. Finally, the trigger was administered 6 h or 24 h after [89Zr]Zr-TCO-Tmab- administration in tumor-bearing mice to liberate the [89Zr]Zr-DFO fragment. PET scans were obtained of tumor-bearing mice that received the trigger 6 h post-[89Zr]Zr-TCO-Tmab administration. Results: The [89Zr]Zr-TCO-Tmab and trigger pair with the best in vivo properties exhibited 83% release in 50% mouse plasma. In tumor-bearing mice the tumor-blood ratios were markedly increased from 1.0 ± 0.4 to 2.3 ± 0.6 (p = 0.0057) and from 2.5 ± 0.7 to 6.6 ± 0.9 (p < 0.0001) when the trigger was administered at 6 h and 24 h post-mAb, respectively. Same day PET imaging clearly showed uptake in the tumor combined with a strongly reduced background due to the fast clearance of the released [89Zr]Zr-DFO-containing fragment from the circulation through the kidneys. Conclusions: This is the first demonstration of the use of trans-cyclooctene-tetrazine click-to-release chemistry to release a radioactive chelator from a mAb in mice to increase tumor-to-blood ratios. Our results suggest that click-cleavable radioimmunoimaging may allow for substantially shorter intervals in PET imaging with full mAbs, reducing radiation doses and potentially even enabling same day imaging.
Assuntos
Neoplasias , Radioimunodetecção , Animais , Camundongos , Trastuzumab , Anticorpos Monoclonais/química , Tomografia por Emissão de Pósitrons/métodos , Ciclo-Octanos/química , Linhagem Celular Tumoral , Zircônio/químicaRESUMO
Radiolabeled antibody treatment with 131I-omburtamab, administered intraventricularly into the cerebrospinal fluid (CSF) space, can deliver therapeutic absorbed doses to sites of leptomeningeal disease. Assessment of distribution and radiation dosimetry is a key element in optimizing such treatments. Using a theranostic approach, we performed pretreatment 131I-omburtamab imaging and dosimetric analysis in patients before therapy. Methods: Whole-body planar images were acquired 3 ± 1, 23 ± 2, and 47 ± 2 h after intracranioventricular administration of 75 ± 5 MBq of 131I-omburtamab via an Ommaya reservoir. Multiple blood samples were also obtained for kinetic analysis. Separate regions of interest (ROIs) were manually drawn to include the lateral ventricles, entire spinal canal CSF space, and over the whole body. Count data in the ROIs were corrected for background and physical decay, converted to activity, and subsequently fitted to an exponential clearance function. The radiation absorbed dose was estimated to the CSF, separately to the spinal column and ventricles, and to the whole body and blood. Biodistribution of the injected radiolabeled antibody was assessed for all patients. Results: Ninety-five patients were included in the analysis. Biodistribution showed prompt localization in the ventricles and spinal CSF space with low systemic distribution, noted primarily as hepatic, renal, and bladder activity after the first day. Using ROI analysis, the effective half-lives were 13 ± 11 h (range, 5-75 h) for CSF in the spinal column, 8 ± 3 h (range, 3-17 h) for ventricles, and 41 ± 11 (range, 23-81 h) for the whole body. Mean absorbed doses were 0.63 ± 0.38 cGy/MBq (range, 0.24-2.25 cGy/MBq) for CSF in the spinal column, 1.03 ± 0.69 cGy/MBq (range, 0.27-5.15 cGy/MBq) for the ventricular CSF, and 0.45 ± 0.32 mGy/MBq (range, 0.05-1.43 mGy/MBq) for the whole body. Conclusion: Pretherapeutic imaging with 131I-omburtamab allows assessment of biodistribution and dosimetry before the administration of therapeutic activity. Absorbed doses to the CSF compartments and whole body derived from the widely applicable serial 131I-omburtamab planar images had acceptable agreement with previously reported data determined from serial 124I-omburtamab PET scans.
Assuntos
Radioimunodetecção , Radiometria , Humanos , Cinética , Distribuição Tecidual , Radiometria/métodos , Anticorpos Monoclonais/uso terapêuticoRESUMO
The principle of pretargeted radioimmunoimaging and therapy has been investigated over the past 30 y in preclinical and clinical settings with the aim of reducing the radiation burden of healthy tissue for antibody-based nuclear medicine techniques. In the past few decades, 4 pretargeting methodologies have been proposed, and 2 of them-the bispecific antibody-hapten and the streptavidin-biotin platforms-have been evaluated in humans in phase 1 and 2 studies. With this review article, we aim to survey clinical pretargeting studies in order to understand the challenges that these platforms have faced in human studies and to provide an overview of how the clinical approval of the pretargeting system has proceeded in the past several decades. Additionally, we will discuss the successes of the pretargeting human studies and compare and highlight the pretargeting approaches and conditions that will advance clinical translation of the pretargeting platform in the future.
Assuntos
Radioimunodetecção , Radioimunoterapia , Humanos , EstreptavidinaRESUMO
Invasive pulmonary aspergillosis (IPA) is a life-threatening lung disease of immunocompromised humans, caused by the opportunistic fungal pathogen Aspergillus fumigatus. Inadequacies in current diagnostic procedures mean that early diagnosis of the disease, critical to patient survival, remains a major clinical challenge, and is leading to the empiric use of antifungal drugs and emergence of azole resistance. A non-invasive procedure that allows both unambiguous detection of IPA and its response to azole treatment is therefore needed. Here, we show that a humanised Aspergillus-specific monoclonal antibody, dual labelled with a radionuclide and fluorophore, can be used in immunoPET/MRI in vivo in a neutropenic mouse model and 3D light sheet fluorescence microscopy ex vivo in the infected mouse lungs to quantify early A. fumigatus lung infections and to monitor the efficacy of azole therapy. Our antibody-guided approach reveals that early drug intervention is critical to prevent complete invasion of the lungs by the fungus, and demonstrates the power of molecular imaging as a non-invasive procedure for tracking IPA in vivo.
Assuntos
Anticorpos Monoclonais Humanizados/imunologia , Antifúngicos/uso terapêutico , Aspergillus fumigatus/imunologia , Pulmão/efeitos dos fármacos , Pulmão/diagnóstico por imagem , Compostos Radiofarmacêuticos/imunologia , Animais , Anticorpos Antifúngicos/química , Anticorpos Antifúngicos/imunologia , Anticorpos Monoclonais Humanizados/química , Aspergillus fumigatus/efeitos dos fármacos , Aspergillus fumigatus/patogenicidade , Azóis/uso terapêutico , Radioisótopos de Cobre/química , Monitoramento de Medicamentos , Corantes Fluorescentes/química , Humanos , Aspergilose Pulmonar Invasiva/diagnóstico por imagem , Aspergilose Pulmonar Invasiva/tratamento farmacológico , Aspergilose Pulmonar Invasiva/microbiologia , Pulmão/microbiologia , Imageamento por Ressonância Magnética , Camundongos , Microscopia de Fluorescência , Tomografia por Emissão de Pósitrons , Radioimunodetecção , Compostos Radiofarmacêuticos/químicaRESUMO
AIMS: In vivo biodistribution of radio labeled ZrO2 nanoparticles is addressed for better imaging, therapy and diagnosis. Nanoparticles are synthesized by microwave assisted sol-gel technique using Fe3O4 as a stabilizer. Antioxidant assay, hemolytic activity in human blood and biodistribution in rabbits was explored to study the therapeutical as well as in vivo targeted diagnostic applications of as synthesized nanoparticles. MAIN METHODS: Fe3O4 stabilized zirconia nanoparticles are synthesized using microwave assisted sol-gel method. Microwave (MW) powers are varied in the range of 100 to 1000 W. As synthesized nanoparticles are evaluated using different characterizations such as X-ray diffractometer, scanning electron microscope, Raman spectroscopy, impedance analyzer, Vickers micro hardness indenter, FTIR, and UV-Vis spectroscopy. In vitro activity of synthesized nanoparticles is checked in freshly extracted human blood serum. To study biodistribution of Fe3O4 stabilized zirconia nanoparticles in rabbit, technetium-99 m was used for labeling purpose. The labeling efficacy and stability of labeled nanoparticles are also measured with instant thin layer chromatography (ITLC) method. Intravenous injection of 99mTc-Fe3O4 stabilized zirconia nanoparticles (0.2 ml), containing 110 MBq of radioactivity, is performed to study the biodistribution; nanoparticles are injected into the ear vein of animal (rabbit). KEY FINDINGS: Zirconia (ZrO2) nanoparticles (NPs) are stabilized using Fe3O4 that were prepared by means of microwave assisted sol-gel method. Crystallite size (~20 nm) agrees well with the values required to stabilize tetragonal zirconia (t-ZrO2). Volume shrinkage results in high value of hardness (~1369). Dielectric constant values, compatible for biomedical application, are observed for tetragonally stabilized samples. Low value of hemolytic response is observed for Fe3O4 stabilized ZrO2 NPs. 99mTc radio labeled ZrO2 NPs proved to be potential candidate to study biodistribution. Biodistribution studies show stability of radiolabeled NPs in the original suspension as well as in blood serum. CT scan of rabbit is performed for several times to check the biodistribution of NPs with time and survival of rabbit. Results suggest that these NPs can also be used as targeted nanoparticles as well as variants of drug payload carrier. SIGNIFICANCE: Results signify that Fe3O4 stabilized ZrO2 nanoparticles synthesized by microwave assisted sol-gel method may be considered as "all-rounder" nanoplatform and are safe enough to be used in diagnostic as well as therapeutic purposes.
Assuntos
Compostos Férricos/metabolismo , Sequestradores de Radicais Livres/metabolismo , Micro-Ondas , Nanopartículas/metabolismo , Radioimunodetecção/métodos , Zircônio/metabolismo , Animais , Compostos Férricos/síntese química , Humanos , Nanopartículas/química , Coelhos , Tecnécio/metabolismo , Distribuição Tecidual/fisiologia , Difração de Raios X/métodos , Zircônio/químicaRESUMO
Radiolabeled leukocyte scans are used in nuclear medicine to detect sites of infection and inflammation. We have previously demonstrated the use of clinical grade immunomagnetic beads to isolate autologous eosinophils and image their distribution in healthy volunteers. Here we describe the use of radiolabeled eosinophils coupled to single-photon emission computed tomography (SPECT) to quantify eosinophil uptake in the lungs of healthy volunteers, patients with asthma, and patients with focal eosinophilic inflammation.
Assuntos
Eosinófilos/fisiologia , Separação Imunomagnética/métodos , Tomografia Computadorizada de Emissão de Fóton Único/métodos , Asma/diagnóstico por imagem , Asma/imunologia , Eosinofilia/diagnóstico por imagem , Humanos , Inflamação/diagnóstico por imagem , Inflamação/imunologia , Cinética , Contagem de Leucócitos , Radioimunodetecção/métodosRESUMO
Background: Prostate cancer is the most common type of cancer for men in many countries. One of the various prostate cancer therapy methods is hormone therapy, and explaining the association between androgen hormones and prostate cancer is a critical role for successful prostate cancer treatment. Materials and Methods: In the current study, the behavior of 3,4-divanillyltetrahydrofuran (DTH) was examined against prostate cancer cells, which have androgen sensitivity differences [LNCaP (+), PC3 (-)]. For this aim, DTH was obtained by extraction of Urtica dioica roots. The molecular structure of isolated compound was confirmed as DTH by liquid chromatography-mass spectrometry and nuclear magnetic resonance spectroscopy analyses. To evaluate the association of androgen sensitivity, DTH was radiolabeled with 131I, and cell uptake assay was performed by using 131I-radiolabeled DTH. Also, cytotoxicity (WST-1) assay of DTH was performed against LNCaP and PC3 cells to determinate the toxic effects of DTH on different androgen mechanisms. Results: The results of assays on cells have shown that DTH lignan behaves different like being more toxic to LNCaP cells than PC3 cells, depending on androgen sensitivity. Conclusion: The results may contribute both the research topics of phytolignan prostate cancer and androgen-sensitive prostate cancer.
Assuntos
Neoplasias da Próstata/induzido quimicamente , Neoplasias da Próstata/radioterapia , Radioimunodetecção/métodos , Urtica dioica/química , Humanos , MasculinoRESUMO
PURPOSE: The aim of the present paper is to review the role of HER2 antibodies, affibodies and nanobodies as vehicles for imaging and therapy approaches in breast cancer, including a detailed look at recent clinical data from antibody drug conjugates and nanobodies as well as affibodies that are currently under development. RESULTS: Clinical and preclinical studies have shown that the use of monoclonal antibodies in molecular imaging is impaired by slow blood clearance, associated with slow and low tumor uptake and with limited tumor penetration potential. Antibody fragments, such as nanobodies, on the other hand, can be radiolabelled with short-lived radioisotopes and provide high-contrast images within a few hours after injection, allowing early diagnosis and reduced radiation exposure of patients. Even in therapy, the small radioactively labeled nanobodies prove to be superior to radioactively labeled monoclonal antibodies due to their higher specificity and their ability to penetrate the tumor. CONCLUSION: While monoclonal antibodies are well established drug delivery vehicles, the current literature on molecular imaging supports the notion that antibody fragments, such as affibodies or nanobodies, might be superior in this approach.
Assuntos
Neoplasias da Mama , Preparações Farmacêuticas , Anticorpos de Domínio Único , Neoplasias da Mama/diagnóstico por imagem , Humanos , Radioimunodetecção , Radioimunoterapia , Receptor ErbB-2RESUMO
OBJECTIVE: Pro-gastrin releasing peptide (ProGRP) plays an oncogenic role in small cell lung cancer (SCLC). The anti-ProGRP(31-98) monoclonal antibody D-D3 can selectively accumulate in SCLC xenografts in nude mice. This study evaluated the effectiveness of a new pretargeting procedure for the early diagnosis of SCLC. METHODS: D-D3 was radiolabeled with technetium-99m (99mTc) using a three-step pretargeting method. Mice with SCLC xenografts were treated with different labeling regimens, and the biodistribution and radioimmunoimaging were explored. The percentage injected dose per gram (%ID/g) in various organs, tumor/non-tumor (T/NT) ratio, and tumor/background (T/B) ratio were also calculated. RESULTS: In vivo distribution experiments revealed that 99mTc-DTPA-biotin was metabolized in the liver and kidney, with rapid elimination in the blood. The T/B ratio was highest in mice treated with biotinylated antibody D-D3 + avidin + 99mTc-DTPA-biotin. Single-photon emission computerized tomography imaging further confirmed that the T/B ratio was highest in this group at all time points. CONCLUSIONS: In contrast to directly labeled D-D3, pretargeting technology displayed specific enhancement and signal amplification in tumors, which could increase the target tumor uptake of 99mTc and provide a new approach for the early diagnosis of SCLC.
Assuntos
Neoplasias Pulmonares , Carcinoma de Pequenas Células do Pulmão , Animais , Biotina , Detecção Precoce de Câncer , Neoplasias Pulmonares/diagnóstico por imagem , Camundongos , Camundongos Nus , Radioimunodetecção , Compostos Radiofarmacêuticos , Carcinoma de Pequenas Células do Pulmão/diagnóstico por imagem , Tecnécio , Pentetato de Tecnécio Tc 99m , Distribuição TecidualRESUMO
Mesothelin is a molecular biomarker of many types of solid cancers, which may represent a highly promising new target in the development of cancer-targeted diagnostic agents. A human anti-mesothelin antibody with a low molecular weight, ET210sc, was applied; this antibody has potent affinity and can penetrate tissue quickly and stably without causing immunoreactions. We developed a new 124/131I-labeled radiotracer of ET210sc. The 124/131I-labeled ET210sc radiotracer showed excellent radiochemical quality (with over 99% radiolabeling yield, 0.07 GBq/µmol specific activity) and remarkable stability in phosphate-buffered saline (>95% at 3 days). The radiotracer retained its potent affinity (dissociation constant, Kd = 0.101 nM). The radiotracer specifically bound to mesothelin-positive cells in vitro. Interestingly, the radiotracer exhibited significant positive-to-negative tumor uptake ratios (1.5:1) 3 days postinjection. The estimated absorbed doses of each organ (e.g., 0.704 mGy/MBq for the rectum; 0.341 mGy/MBq for the spleen) met the medical safety standards for further clinical applications. Our findings provide an initial proof of concept for the potential use of 124/131I-labeled ET210sc radiotracers to detect mesothelin-overexpressing cancer. 124I-ET210sc is proposed to be an ideal imaging agent for further clinical applications.
Assuntos
Proteínas Ligadas por GPI/metabolismo , Neoplasias/diagnóstico por imagem , Animais , Western Blotting , Linhagem Celular Tumoral , Feminino , Citometria de Fluxo , Humanos , Imuno-Histoquímica , Isótopos de Iodo/análise , Mesotelina , Camundongos Endogâmicos BALB C , Tomografia por Emissão de Pósitrons , Radioimunodetecção , RadiometriaAssuntos
Artrite/diagnóstico por imagem , Doenças da Medula Óssea/diagnóstico por imagem , Edema/diagnóstico por imagem , Radioimunodetecção/métodos , Articulação Sacroilíaca/patologia , Sacroileíte/diagnóstico por imagem , Adulto , Certolizumab Pegol , Feminino , Humanos , Imageamento por Ressonância Magnética , Masculino , Tomografia por Emissão de Pósitrons combinada à Tomografia Computadorizada , Traçadores Radioativos , Articulação Sacroilíaca/diagnóstico por imagem , TecnécioRESUMO
The immune system's role in atherosclerosis has long been an important research topic and is increasingly investigated for therapeutic and diagnostic purposes. Therefore, noninvasive imaging of hematopoietic organs and immune cells will undoubtedly improve atherosclerosis phenotyping and serve as a monitoring method for immunotherapeutic treatments. Among the available imaging techniques, positron emission tomography's unique features make it an ideal tool to quantitatively image the immune response in the context of atherosclerosis and afford reliable readouts to guide medical interventions in cardiovascular disease. Here, we summarize the state of the art in the field of atherosclerosis positron emission tomography immunoimaging and provide an outlook on current and future applications.
Assuntos
Aterosclerose/diagnóstico por imagem , Aterosclerose/imunologia , Tomografia por Emissão de Pósitrons/métodos , Aterosclerose/metabolismo , Sistema Hematopoético/diagnóstico por imagem , Humanos , Nanopartículas , Fagócitos , Placa Aterosclerótica/diagnóstico por imagem , Radioimunodetecção , Compostos RadiofarmacêuticosRESUMO
CONTEXT: Preliminary results have demonstrated that prostate-specific membrane antigen (PSMA) is highly expressed on the cell surface of the microvasculature of several solid tumors, including renal cell carcinoma (RCC). OBJECTIVE: To assess the role of PSMA positron emission tomography (PET)/computed tomography (CT) in RCC patients and to discuss its possible inclusion in the clinical management of these patients. EVIDENCE ACQUISITION: A systematic literature search was performed for studies on PET/CT in patients with RCC up to April 2018. MEDLINE databases, such as Pubmed, Web of Science, and Scopus were consulted using the following keywords: "Renal Cell Carcinoma" AND "PET/CT", "Renal Cancer" AND "PET/CT", "renal cancer" AND "PSMA PET/CT", and "renal cell carcinoma" AND "PSMA PET/CT". EVIDENCE SYNTHESIS: Thirteen articles were retrieved from the available literature; the majority of them were relative to metastatic RCC (n=11/13, 85%) and eight of them were clinical cases, thus providing low-quality data. No diagnostic benefit of PSMA PET/CT was found in the evaluation of primary tumors. PSMA PET/CT may be a useful imaging modality in whole-body staging and restaging of patients with RCC and in the assessment of lesions that remained unclear on conventional imaging. Furthermore, PSMA PET/CT may predict the response to anti-angiogenic-targeted therapies. CONCLUSIONS: The utility of PSMA-based PET imaging in the assessment of patients with RCC is encouraging. However, the available preliminary results will need to be addressed with larger prospective trials. PATIENT SUMMARY: In this mini-review, we evaluated the usefulness of an alternative imaging technique for patients with renal cell cancer. We found that this new imaging modality may indeed be useful. We conclude that the preliminary data should be assessed by larger studies.
Assuntos
Antígenos de Superfície , Carcinoma de Células Renais/diagnóstico por imagem , Glutamato Carboxipeptidase II , Neoplasias Renais/diagnóstico por imagem , Tomografia por Emissão de Pósitrons combinada à Tomografia Computadorizada/métodos , Humanos , RadioimunodetecçãoRESUMO
Triple-negative breast cancer (TNBC) is a highly aggressive tumour that lacks marker for targeted diagnosis. Recently, it was reported that toll-like receptor 5 (TLR5) was associated with some kind of tumours, especially in TNBC, but whether it could be used as a non-invasive monitoring target is not fully understood. Here, we established TLR5- 4T1 cell line with lentivirus-shRNA-TLR5 knock-down transfection (with tag GFP, green fluorescent protein, TLR5- 4T1) and control TLR5+ 4T1 cell line with negative control lentivirus transfection. The effect of TLR5 down-regulation was detected with qPCR and Western blot. 125 I-anti-TLR5 mAb and control isotype 125 I-IgG were prepared and injected to TLR5+/- 4T1-bearing mice models, respectively. Whole-body phosphor-autoradiography, fluorescence imaging and biodistribution were performed. Furthermore, ex vivo tumour TLR5 expression was proved through immunohistochemistry staining. We found that 125 I-anti-TLR5 mAb could bind to TLR5+ 4T1 with high affinity and specificity. Whole-body phosphor-autoradiography after 125 I-anti-TLR5 mAb injection showed TLR5+ 4T1 tumour images in 24 hours, more clearly in 48 hours. Radioactivities in tumour tissues were positively related with TLR5 expression. Biodistribution assay showed that 125 I-anti-TLR5 mAb was mainly metabolized through the liver and kidney, and 125 I-anti-TLR5 mAb was much more accumulated in TLR5+ 4T1 tumour than TLR5- 4T1. In vivo fluorescence imaging successfully showed tumour tissues clearly both in TLR5+ and TLR5- 4T1 mice compared with lentivirus untreated 4T1 tumour. Immunohistochemistry staining showed that TLR5 expression in tumours was indeed down-regulated in TLR5- 4T1 mice. Our results indicated that 125 I-antiTLR5 mAb was an ideal agent for non-invasive imaging of TLR5+ tumours; TLR5 may be as a novel molecular target for TNBC non-invasive diagnosis.
Assuntos
Biomarcadores Tumorais/metabolismo , Neoplasias Mamárias Experimentais/metabolismo , Radioimunodetecção/métodos , Receptor 5 Toll-Like/metabolismo , Neoplasias de Mama Triplo Negativas/metabolismo , Animais , Anticorpos Monoclonais/análise , Anticorpos Monoclonais/imunologia , Autorradiografia/métodos , Biomarcadores Tumorais/genética , Linhagem Celular Tumoral , Feminino , Proteínas de Fluorescência Verde/genética , Proteínas de Fluorescência Verde/metabolismo , Imuno-Histoquímica/métodos , Radioisótopos do Iodo/metabolismo , Radioisótopos do Iodo/farmacocinética , Masculino , Neoplasias Mamárias Experimentais/diagnóstico , Neoplasias Mamárias Experimentais/genética , Camundongos Nus , Interferência de RNA , Coloração e Rotulagem/métodos , Distribuição Tecidual , Receptor 5 Toll-Like/genética , Neoplasias de Mama Triplo Negativas/diagnóstico , Neoplasias de Mama Triplo Negativas/genéticaRESUMO
This study was to evaluate the potential of 99mTc-Hynic-fragments of Rituximab as radioimmunoscintigraphic agents for diagnosis of patients with Non-Hodgkin's Lymphoma (NHL). Rituximab was digested with immobilized pepsin and papain to yield F(ab')2-Rituximab and F(ab')-Rituximab fragments respectively. Purified fragments were characterized by SE-HPLC and SDS-PAGE and subsequently radiolabeled with technetium-99m using Hynic as bifunctional chelator. The 99mTc-Hynic-F(ab')2-Rituximab and 99mTc-Hynic-F(ab')-Rituximab exhibited good in-vitro stability and specificity to Raji cells. Biodistribution studies demonstrated rapid pharmacokinetics and clearance predominantly through renal route.
Assuntos
Antineoplásicos/uso terapêutico , Linfoma não Hodgkin/tratamento farmacológico , Radioimunodetecção/métodos , Rituximab/uso terapêutico , Tecnécio/administração & dosagem , Animais , Humanos , Camundongos , Rituximab/imunologiaRESUMO
BACKGROUND: Pancreatic ductal adenocarcinoma (PDAC) accounts for >90% of pancreatic malignancies, and has median survival of <6 months. There is an urgent need for diagnostic and therapeutic options for PDAC. Centrin1 (CETN1) is a novel member of Cancer/Testis Antigens, with a 25-fold increase of CETN1 gene expression in PDX from PDAC patients. The absence of selective anti-CETN1 antibodies is hampering CETN1 use for diagnosis and therapy. Here we report the generation of highly specific for CETN1 antibodies and their evaluation for radioimmunoimaging and radioimmunotherapy (RIT) of experimental PDAC. METHODS: The antibodies to CETN1 were generated via mice immunization with immunogenic peptide distinguishing CETN1 from CETN2. Patient tumor microarrays were used to evaluate the binding of the immune serum to PDAC versus normal pancreas. The antibodies were tested for their preferential binding to CETN1 over CETN2 by ELISA. Mice bearing PDAC MiaPaCa2 xenografts were imaged with microSPECT/CT and treated with 213 Bi- and 177 Lu-labeled antibodies to CETN1. RESULTS: Immune serum bind to 50% PDAC cases on patient tumor microarrays with no specific binding to normal pancreas. Antibodies demonstrated preferential binding to CETN1 versus CETN2. Antibody 69-11 localized to PDAC xenografts in mice in vivo and ex vivo. RIT of PDAC xenografts with 213 Bi-labeled antibodies was effective, safe, and CETN1-specific. CONCLUSIONS: The results demonstrate the ability of these novel antibodies to detect CETN1 both in vitro and in vivo; as well, the RIT treatment of experimental PDAC when radiolabeled with 213 Bi is highly efficient and safe. Further evaluation of these novel reagents for diagnosis and treatment of PDAC is warranted.
Assuntos
Anticorpos , Antígenos de Neoplasias , Proteínas de Membrana , Imagem Molecular , Neoplasias Pancreáticas/diagnóstico , Neoplasias Pancreáticas/terapia , Radioimunodetecção , Radioimunoterapia , Sequência de Aminoácidos , Animais , Antígenos de Neoplasias/química , Antígenos de Neoplasias/metabolismo , Linhagem Celular Tumoral , Modelos Animais de Doenças , Feminino , Humanos , Imunoglobulina G/imunologia , Imunoglobulina M/imunologia , Masculino , Proteínas de Membrana/antagonistas & inibidores , Proteínas de Membrana/química , Proteínas de Membrana/metabolismo , Camundongos , Imagem Molecular/métodos , Neoplasias Pancreáticas/etiologia , Radioimunodetecção/métodos , Radioimunoterapia/métodos , Tomografia Computadorizada com Tomografia Computadorizada de Emissão de Fóton Único , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
Human carbonic anhydrase IX is a membrane enzyme that is significantly expressed in some types of cancer cells, while copper radioisotopes offer wide range of diagnostic, therapeutic and theranostic properties. The work was focused on a new approach to the labelling of antibody IgG M75 for epitope human carbonic anhydrase IX with copper radioisotopes 61Cu and 64Cu and its in vivo testing in mice with inoculated colorectal cancer. Monoclonal antibody IgG M75 for epitope human carbonic anhydrase IX was successfully conjugated with copper-specific chelator "phosphinate" and labelled with 61Cu and 64Cu The obtained molecule has considerable potential as a radioimmuno pharmaceutical suitable for imaging of tumours expressing carbonic anhydrase IX by positron emission tomography (PET).
Assuntos
Anticorpos Monoclonais/química , Antígenos de Neoplasias/imunologia , Anidrase Carbônica IX/imunologia , Radioisótopos de Cobre/química , Animais , Anticorpos Monoclonais/farmacocinética , Neoplasias Colorretais/diagnóstico por imagem , Neoplasias Colorretais/enzimologia , Radioisótopos de Cobre/farmacocinética , Células HT29 , Humanos , Imunoconjugados/química , Imunoconjugados/farmacocinética , Imunoglobulina G/química , Masculino , Camundongos , Camundongos Nus , Tomografia por Emissão de Pósitrons , Radioimunodetecção , Compostos Radiofarmacêuticos/química , Compostos Radiofarmacêuticos/farmacocinética , Distribuição TecidualRESUMO
Recently, inhibiting the PD-1/PD-L1 checkpoint pathway utilizing anti-PD-1 or anti-PD-L1 antibodies has achieved great clinical success in cancer treatment. However, anti-PD-1 immunotherapy cannot be applied to all cancer patients, no more than 25% showed a positive response. Immunohistochemistry (IHC) is the gold standard to determine the PD-L1 expression level in malignant lesions, but a noninvasive imaging-meditated strategy is urgently required for clinical diagnosis to cover the shortcomings of invasive techniques. MX001, which is an anti-PD-L1 antibody, was labeled with Cu-64 ( t1/2 = 12.7 h) and purified by PD-10 chromatography. Comprehensive studies including positron emission tomography (PET), ex vivo biodistribution, IHC, and immunotherapy have been performed in mice bearing MC38 (PD-L1 positive (+)) and 4T1 (PD-L1 negative (-)) xenografts. PET imaging of [18F]FDG was taken before and after therapy to monitor the therapeutic efficacy. [64Cu]Cu-NOTA-MX001 exhibited 2.3 ± 1.2, 5.6 ± 2.1, 5.6 ± 1.2, 6.1 ± 1.1, 6.1 ± 0.5, and 10.2 ± 1.7%ID/g uptake in MC38 xenografts at 0.5, 12, 24, 36, 48, and 62 h post-injection (p.i.), respectively. Meanwhile, the uptake in the liver and muscle at corresponding time points was 17.5 ± 2.2, 8.4 ± 2.4, 11.3 ± 3.2, 7.2 ± 2.1, 7.9.1 ± 3.5, and 3.8 ± 1.8%ID/g, and 1.2 ± 0.5, 1.3 ± 0.4, 1.5 ± 0.5, 0.7 ± 0.1, 0.6 ± 0.2, and 0.2 ± 0.1%ID/g, respectively. The uptake of [18F]FDG in MC38 and 4T1 xenografts at 1-h p.i. was 5.3 ± 0.4 and 6.4 ± 0.6%ID/g, while the uptake of [64Cu]Cu-NOTA-MX001 was 5.6 ± 0.3 and 1.3 ± 0.4%ID/g at 12-h p.i. IHC analysis confirmed that the MC38 tumor exhibited high PD-L1 expression, and the 4T1 tumor, liver, and muscle exhibited low PD-L1 expression. In addition, MC38 xenografts were suppressed by MX001 about 88% in the immunotherapy study. MX001 was successfully developed as a fully human anti-PD-L1 antibody with a high binding affinity in mouse, monkey, and human. The in vivo pharmacokinetics of MX001 was evaluated with PET imaging after being radiolabeled with Cu-64. The uptake of [64Cu]Cu-NOTA-MX001 was clearly correlated to the PD-L1 expression on various types of cancer. Subsequent immunotherapy studies demonstrated that MX001 could effectively suppress tumor growth with positive PD-L1 expression, but had poor antitumor efficacy on tumors which exhibited low PD-L1 expression. Together with the above results, MX001 has the potential to be further developed as an antibody theranostic agent for both PET imaging and immunotherapy of cancers in clinics.
Assuntos
Anticorpos/uso terapêutico , Antígeno B7-H1/metabolismo , Imunoterapia/métodos , Animais , Antígeno B7-H1/imunologia , Linhagem Celular Tumoral , Feminino , Humanos , Imuno-Histoquímica , Camundongos , Tomografia por Emissão de Pósitrons , Receptor de Morte Celular Programada 1/imunologia , Receptor de Morte Celular Programada 1/metabolismo , RadioimunodetecçãoRESUMO
PURPOSE: This study aimed to investigate theranostic strategies in colorectal and skin cancer based on fragments of cetuximab, an anti-EGFR mAb, labeled with radionuclide with imaging and therapeutic properties, 111In and 177Lu, respectively. METHODS: We designed F(ab')2-fragments of cetuximab radiolabeled with 111In and 177Lu. 111In-F(ab')2-cetuximab tumor targeting and biodistribution were evaluated by SPECT in BalbC nude mice bearing primary colorectal tumors. The efficacy of 111In-F(ab')2-cetuximab to assess therapy efficacy was performed on BalbC nude mice bearing colorectal tumors receiving 17-DMAG, an HSP90 inhibitor. Therapeutic efficacy of the radioimmunotherapy based on 177Lu-F(ab')2-cetuximab was evaluated in SWISS nude mice bearing A431 tumors. RESULTS: Radiolabeling procedure did not change F(ab')2-cetuximab and cetuximab immunoreactivity nor affinity for HER1 in vitro. 111In-DOTAGA-F(ab')2-cetuximab exhibited a peak tumor uptake at 24 h post-injection and showed a high tumor specificity determined by a significant decrease in tumor uptake after the addition of an excess of unlabeled-DOTAGA-F(ab')2-cetuximab. SPECT imaging of 111In-DOTAGA-F(ab')2-cetuximab allowed an accurate evaluation of tumor growth and successfully predicted the decrease in tumor growth induced by 17-DMAG. Finally, 177Lu-DOTAGA-F(ab')2-cetuximab radioimmunotherapy showed a significant reduction of tumor growth at 4 and 8 MBq doses. CONCLUSIONS: 111In-DOTAGA-F(ab')2-cetuximab is a reliable and stable tool for specific in vivo tumor targeting and is suitable for therapy efficacy assessment. 177Lu-DOTAGA-F(ab')2-cetuximab is an interesting theranostic tool allowing therapy and imaging.