RESUMO
Lymphocyte activation gene 3 (LAG-3) has attracted much attention as a potentially valuable immune checkpoint. Individual identification of LAG-3 expression at screening and during treatment could improve the successful implementation of anti-LAG-3 therapies. HuL13 is a human IgG1 monoclonal antibody that binds to the LAG-3 receptor in T cells. Here, we used [89Zr]Zr-labeled HuL13 to delineate LAG-3+ T-cell infiltration into tumors via positron emission tomography (PET) imaging. A549/LAG-3 cells, which stably express LAG-3, were generated by infection with lentivirus. The uptake of [89Zr]Zr-DFO-HuL13 in A549/LAG-3 cells was greater than that in the negative control (A549/NC) cells at each time point. The equilibrium dissociation constant (Kd) of [89Zr]Zr-DFO-HuL13 for the LAG-3 receptor was 8.22 nM. PET imaging revealed significant uptake in the tumor areas of A549/LAG-3 tumor-bearing mice from 24 h after injection (SUVmax = 2.43 ± 0.06 at 24 h). As a proof of concept, PET imaging of the [89Zr]Zr-DFO-HuL13 tracer was further investigated in an MC38 tumor-bearing humanized LAG-3 mouse model. PET imaging revealed that the [89Zr]Zr-DFO-HuL13 tracer specifically targets human LAG-3 expressed on tumor-infiltrating lymphocytes (TILs). In addition to the tumors, the spleen was also noticeably visible. Tumor uptake of the [89Zr]Zr-DFO-HuL13 tracer was lower than its uptake in the spleen, but high uptake in the spleen could be reduced by coinjection of unlabeled antibodies. Coinjection of unlabeled antibodies increases tracer activity in the blood pool, thereby improving tumor uptake. Dosimetry evaluation of the healthy mouse models revealed that the highest absorbed radiation dose was in the spleen, followed by the liver and heart wall. In summary, these studies demonstrate the feasibility of using the [89Zr]Zr-DFO-HuL13 tracer for the detection of LAG-3 expression on TILs. Further clinical evaluation of the [89Zr]Zr-DFO-HuL13 tracer may be of significant help in the stratification and management of patients suitable for anti-LAG-3 therapy.
Assuntos
Proteína do Gene 3 de Ativação de Linfócitos , Linfócitos do Interstício Tumoral , Tomografia por Emissão de Pósitrons , Zircônio , Animais , Humanos , Camundongos , Zircônio/química , Linfócitos do Interstício Tumoral/imunologia , Linfócitos do Interstício Tumoral/metabolismo , Tomografia por Emissão de Pósitrons/métodos , Linhagem Celular Tumoral , Antígenos CD/metabolismo , Antígenos CD/imunologia , Radioisótopos/química , Anticorpos Monoclonais/química , Feminino , Distribuição TecidualRESUMO
Carbonic anhydrase IX (CAIX), a zinc metal transmembrane protein, is highly expressed in 95% of clear cell renal cell carcinomas (ccRCCs). A positron emission tomography (PET) probe designed to target CAIX in nuclear medicine imaging technology can achieve precise positioning, is noninvasive, and can be used to monitor CAIX expression in lesions in real time. In this study, we constructed a novel acetazolamide dual-targeted small-molecule probe [68Ga]Ga-LF-4, which targets CAIX by binding to a specific amino acid sequence. After attenuation correction, the radiolabeling yield reached 66.95 ± 0.57% (n = 5) after 15 min of reaction and the radiochemical purity reached 99% (n = 5). [68Ga]Ga-LF-4 has good in vitro and in vivo stability, and in vivo safety and high affinity for CAIX, with a Kd value of 6.62 nM. Moreover, [68Ga]Ga-LF-4 could be quickly cleared from the blood in vivo. The biodistribution study revealed that the [68Ga]Ga-LF-4 signal was concentrated in the heart, lung, and kidney after administration, which was the same as that observed in the micro-PET/CT study. In a ccRCC patient-derived xenograft (PDX) model, the signal significantly accumulated in the tumor after administration, where it was retained for up to 4 h. After competitive blockade with LF-4, uptake at the tumor site was significantly reduced. The SUVmax of the probe [68Ga]Ga-LF-4 at the ccRCC tumor site was three times greater than that in the PC3 group with low CAIX expression at 30 min (ccRCC vs PC3:1.86 ± 0.03 vs 0.62 ± 0.01, t = 48.2, P < 0.0001). These results indicate that [68Ga]Ga-LF-4 is a novel small-molecule probe that targets CAIX and can be used to image localized and metastatic ccRCC lesions.
Assuntos
Anidrase Carbônica IX , Carcinoma de Células Renais , Radioisótopos de Gálio , Neoplasias Renais , Animais , Anidrase Carbônica IX/metabolismo , Anidrase Carbônica IX/antagonistas & inibidores , Humanos , Camundongos , Carcinoma de Células Renais/diagnóstico por imagem , Carcinoma de Células Renais/metabolismo , Neoplasias Renais/diagnóstico por imagem , Neoplasias Renais/metabolismo , Distribuição Tecidual , Linhagem Celular Tumoral , Compostos Radiofarmacêuticos/farmacocinética , Compostos Radiofarmacêuticos/química , Camundongos Nus , Antígenos de Neoplasias/metabolismo , Sondas Moleculares/farmacocinética , Sondas Moleculares/química , Tomografia por Emissão de Pósitrons combinada à Tomografia Computadorizada/métodos , Acetazolamida/farmacocinética , Feminino , Camundongos Endogâmicos BALB C , Tomografia por Emissão de Pósitrons/métodos , Masculino , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
T cell immunoglobulin and mucin domain-3 (TIM3; HAVCR2) is a transmembrane protein that exerts negative regulatory control over T cell responses. Studies have demonstrated an upregulation of TIM3 expression in tumor-infiltrating lymphocytes (TILs) in cancer patients. In this investigation, a series of monoclonal antibodies targeting TIM3 were produced by hybridoma technology. Among them, C23 exhibited favorable biological properties. To enable specific binding, we developed a 124I/125I-C23 radio-tracer via N-bromosuccinimide (NBS)-mediated labeling of the monoclonal antibody C23. Binding affinity and specificity were assessed using the 293T-TIM3 cell line, which overexpresses TIM3, and the parent 293T cells. Furthermore, biodistribution and in vivo imaging of 124I/125I-C23 were examined in HEK293TIM3 xenograft models and allograft models of 4T1 (mouse breast cancer cells) and CT26 (mouse colon cancer cells). Micro-PET/CT imaging was conducted at intervals of 4, 24, 48, 72, and/or 96 h post intravenous administration of 3.7-7.4 MBq 124I-C23 in the respective model mice. Additionally, immunohistochemistry (IHC) staining of TIM3 expression in dissected tumor organs was performed, along with an assessment of the corresponding expression of Programmed Death 1 (PD1), CD3, and CD8 in the tumors. The C23 monoclonal antibody (mAb) specifically binds to TIM3 protein with a dissociation constant of 23.28 nM. The 124I-C23 and 125I-C23 radio-tracer were successfully prepared with a labeling yield of 83.59 ± 0.35% and 92.35 ± 0.20%, respectively, and over 95.00% radiochemical purity. Stability results indicated that the radiochemical purity of 124I/125I-C23 in phosphate-buffered saline (PBS) and 5% human serum albumin (HSA) was still >80% after 96 h. 125I-C23 uptake in 293T-TIM3 cells was 2.80 ± 0.12%, which was significantly higher than that in 293T cells (1.08 ± 0.08%), and 125I-C23 uptake by 293T-TIM3 cells was significantly blocked at 60 and 120 min in the blocking groups. Pharmacokinetics analysis in vivo revealed an elimination time of 14.62 h and a distribution time of 0.4672 h for 125I-C23. Micro-PET/CT imaging showed that the 124I-C23 probe uptake in the 293T-TIM3 model significantly differed from that of the negative control group and blocking group. In the humanized mouse model, the 124I-C23 probe had obvious specific uptake in the 4T1 and CT26 models and maximum uptake at 24 h in tumor tissues (SUVmax (the maximum standardized uptake value) in 4T1 and CT26 humanized TIM3 murine tumor models: 0.59 ± 0.01 and 0.76 ± 0.02, respectively). Immunohistochemistry of tumor tissues from these mouse models showed comparable TIM3 expression. CD3 and CD8 cells and PD-1 expression were also observed in TIM3-expressing tumor tissues. The TIM3-targeting antibody C23 showed good affinity and specificity. The 124I/125I-C23 probe has obvious targeting specificity for TIM3 in vitro and in vivo. Our results suggest that 124I/125I-C23 is a promising tracer for TIM3 imaging and may have great potential in monitoring immune checkpoint drug efficacy.
Assuntos
Anticorpos Monoclonais , Neoplasias , Animais , Humanos , Camundongos , Anticorpos Monoclonais/química , Linhagem Celular Tumoral , Receptor Celular 2 do Vírus da Hepatite A/metabolismo , Radioisótopos do Iodo , Tomografia por Emissão de Pósitrons combinada à Tomografia Computadorizada , Compostos Radiofarmacêuticos/farmacocinética , Distribuição TecidualRESUMO
BACKGROUND: Learning to perform strabismus surgery is an essential aspect of ophthalmologists' surgical training. Automated classification strategy for surgical steps can improve the effectiveness of training curricula and the efficient evaluation of residents' performance. To this end, we aimed to develop and validate a deep learning (DL) model for automated detecting strabismus surgery steps in the videos. METHODS: In this study, we gathered 479 strabismus surgery videos from Shanghai Children's Hospital, affiliated to Shanghai Jiao Tong University School of Medicine, spanning July 2017 to October 2021. The videos were manually cut into 3345 clips of the eight strabismus surgical steps based on the International Council of Ophthalmology's Ophthalmology Surgical Competency Assessment Rubrics (ICO-OSCAR: strabismus). The videos dataset was randomly split by eye-level into a training (60%), validation (20%) and testing dataset (20%). We evaluated two hybrid DL algorithms: a Recurrent Neural Network (RNN) based and a Transformer-based model. The evaluation metrics included: accuracy, area under the receiver operating characteristic curve, precision, recall and F1-score. RESULTS: DL models identified the steps in video clips of strabismus surgery achieved macro-average AUC of 1.00 (95% CI 1.00-1.00) with Transformer-based model and 0.98 (95% CI 0.97-1.00) with RNN-based model, respectively. The Transformer-based model yielded a higher accuracy compared with RNN-based models (0.96 vs. 0.83, p < 0.001). In detecting different steps of strabismus surgery, the predictive ability of the Transformer-based model was better than that of the RNN. Precision ranged between 0.90 and 1 for the Transformer-based model and 0.75 to 0.94 for the RNN-based model. The f1-score ranged between 0.93 and 1 for the Transformer-based model and 0.78 to 0.92 for the RNN-based model. CONCLUSION: The DL models can automate identify video steps of strabismus surgery with high accuracy and Transformer-based algorithms show excellent performance when modeling spatiotemporal features of video frames.
Assuntos
Aprendizado Profundo , Músculos Oculomotores , Procedimentos Cirúrgicos Oftalmológicos , Estrabismo , Gravação em Vídeo , Humanos , Estrabismo/cirurgia , Músculos Oculomotores/cirurgia , Oftalmologia/educação , Curva ROC , Competência Clínica , Redes Neurais de Computação , Algoritmos , Internato e Residência , Educação de Pós-Graduação em Medicina/métodosRESUMO
Coronavirus disease 2019 (COVID-19), which is caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection, poses a significant threat to public health. Angiotensin-converting enzyme 2 (ACE2) is a key receptor for SARS-CoV-2 infection. Recombinant human ACE2 (RhACE2), as a soluble supplement for human ACE2, can competitively block SARS-CoV-2 infection. In this study, a mouse organ in situ rhACE2 high aggregation model was constructed for the first time, and in vivo real-time positron emission tomography (PET) imaging of rhACE2 in the mouse model was performed using an ACE2-specific agent 68 Ga-HZ20. This radiotracer exhibits reliable radiochemical properties in vitro and maintains a high affinity for rhACE2 in vivo. In terms of probe uptake, 68 Ga-HZ20 showed a good target-to-nontarget ratio and was rapidly cleared from the circulatory system and excreted by the kidneys and urinary system. PET imaging with this radiotracer can noninvasively and accurately monitor the content and distribution of rhACE2 in the body, which clarifies that rhACE2 can aggregate in multiple organs, suggesting the preventive and therapeutic potential of rhACE2 for SARS-CoV-2 and COVID-19.
Assuntos
COVID-19 , SARS-CoV-2 , Humanos , Animais , Camundongos , COVID-19/diagnóstico por imagem , Enzima de Conversão de Angiotensina 2 , Tomografia por Emissão de Pósitrons combinada à Tomografia Computadorizada , Peptidil Dipeptidase A , Modelos Animais de DoençasRESUMO
PURPOSE: Programmed cell death protein-1/ligand-1 (PD-1/L1) blockade has been a breakthrough in the treatment of patients with non-small cell lung cancer (NSCLC), but there is still a lack of effective methods to screen patients. Here we report a novel 68 Ga-labeled nanobody [68 Ga]Ga-THP-APN09 for PET imaging of PD-L1 status in mouse models and a first-in-human study in NSCLC patients. METHODS: [68 Ga]Ga-THP-APN09 was prepared by site-specific radiolabeling, with no further purification. Cell uptake assays were completed in the human lung adenocarcinoma cell line A549, NSCLC cell line H1975 and human PD-L1 gene-transfected A549 cells (A549PD-L1). The imaging to image PD-L1 status and biodistribution were investigated in tumor-bearing mice of these three tumor cell types. The first-in-human clinical translational trial was registered as NCT05156515. The safety, radiation dosimetry, biodistribution, and correlations of tracer uptake with immunohistochemical staining and major pathologic response (MPR) were evaluated in NSCLC patients who underwent adjuvant immunotherapy combined with chemotherapy. RESULTS: Radiosynthesis of [68 Ga]Ga-THP-APN09 was achieved at room temperature and a pH of 6.0-6.5 in 10 min with a high radiochemical yield (> 99%) and 13.9-27.8 GBq/µmol molar activity. The results of the cell uptake study reflected variable levels of surface PD-L1 expression observed by flow cytometry in the order A549PD-L1 > H1975 > A549. In small-animal PET/CT imaging, H1975 and A549PD-L1 tumors were clearly visualized in an 8.3:1 and 2.2:1 ratios over PD-L1-negative A549 tumors. Ex vivo biodistribution studies showed that tumor uptake was consistent with the PET results, with the highest A549PD-L1 being taken up the most (8.20 ± 0.87%ID/g), followed by H1975 (3.69 ± 0.50%ID/g) and A549 (0.90 ± 0.16%ID/g). Nine resectable NSCLC patients were enrolled in the clinical study. Uptake of [68 Ga]Ga-THP-APN09 was mainly observed in the kidneys and spleen, followed by low uptake in bone marrow. The radiation dose is within a reliable range. Tumor uptake was positively correlated with PD-L1 expression TPS (rs = 0.8763, P = 0.019). Tumor uptake of [68 Ga]Ga-THP-APN09 (SUVmax) in MPR patients was higher than that in non-MPR patients (median SUVmax 2.73 vs. 2.10, P = 0.036, determined with Mann-Whitney U-test). CONCLUSION: [68 Ga]Ga-THP-APN09 has the potential to be transformed into a kit-based radiotracer for rapid, simple, one-step, room temperature radiolabeling. The tracer can detect PD-L1 expression levels in tumors, and it may make it possibility to predict the response of PD-1 immunotherapy combined with chemotherapy. Confirmation in a large number of cases is needed. TRIAL REGISTRATION: Clinical Trial (NCT05156515). Registered 12 December 2021.
Assuntos
Carcinoma Pulmonar de Células não Pequenas , Neoplasias Pulmonares , Humanos , Camundongos , Animais , Tomografia por Emissão de Pósitrons combinada à Tomografia Computadorizada/métodos , Carcinoma Pulmonar de Células não Pequenas/diagnóstico por imagem , Carcinoma Pulmonar de Células não Pequenas/terapia , Radioisótopos de Gálio , Antígeno B7-H1/metabolismo , Distribuição Tecidual , Receptor de Morte Celular Programada 1/metabolismo , Neoplasias Pulmonares/diagnóstico por imagem , Tomografia por Emissão de Pósitrons/métodos , Linhagem Celular TumoralRESUMO
Activated T cells played critical roles in immunotherapy and adoptive T cell therapy, and a non-invasive imaging strategy can provide us useful information concerning the transportation, accumulation, and homing of T cells in vivo. In this paper, by utilizing the long half-life radionuclide iodine-124 (124I) and CD25 specific monoclonal antibody Basiliximab, we have fabricated a novel probe, namely, 124I-Basiliximab, which was highly promising in the immuno-PET imaging of T cells. In vitro, 124I-Basiliximab had superior affinity to CD25 protein (Kd = 5.31 nM) and exhibited much higher accumulation in CD25 high-expression lymphoma cell line Karpas299 than that in CD25-negative cell line Daudi. In vivo, 124I-Basiliximab was excreted slowly from the body of mice, rendering it a relatively high effective dose (0.393 mSv/MBq) when applied in the immuno-PET imaging. In Karpas299 tumor xenograft, 124I-Basiliximab probe was observed to accumulate in the tumor quickly after tracer administration, with the optimal image acquired at 24 h post-injection. More importantly, PHA-activated hPBMC had much higher uptake of 124I-Basiliximab, indicating the potential utility of 124I-Basiliximab to discriminate activated hPBMC from its non-activated status. In summary, 124I-Basiliximab was fabricated for the first time, which can be applied in CD25-targeted immuno-PET imaging of activated T cells in vivo.
Assuntos
Neoplasias , Linfócitos T , Animais , Basiliximab , Humanos , Radioisótopos do Iodo , Camundongos , Tomografia por Emissão de Pósitrons , Proteínas Recombinantes de FusãoRESUMO
Mesothelin (MSLN) is a molecular biomarker of many types of solid tumors, such as mesothelioma, pancreatic cancer, and colon cancer. Owing to the significant difference in expression between cancer cells and normal cells, mesothelin has been widely used as a key target in cancer immunotherapy. In this study, we used iodine isotope (nat/124/125I)-labeled mesothelin antibodies to noninvasively detect MSLN expression in mice with LS174T colon cancer. The 124I-labeled MSLN antibody showed a high radiochemical purity (RCP, >99%) and specific activity (20.8-67.8 GBq/µmol) after purification and was stable in 5% HSA and PBS (>95% RCP at 8 days). Western blot analysis indicated that the LS174T cells showed a higher MSLN protein level than the HepG2 cells. The half maximal effective concentration (EC50) values of the MSLN antibody and natI-anti-MSLN were 34.77 ± 3.72 ng/mL and 32.60 ± 2.52 ng/mL (P = 0.63), respectively. The dissociation constant of 124I-anti-MSLN binding to MSLN protein was 16.0 nM. The radiotracer showed a significantly higher uptake in LS174T cells than in HepG2 tumor cells (1.56 ± 0.09 vs 0.81 ± 0.03, P = 0.0016) 2 days postinjection. The LS174T mouse models showed extremely low organ uptake and high tumor uptake 96 h after the injection of 124I-anti-MSLN, and the T/M values were much higher than those of the other imaging groups (10.56 ± 1.20 for 124I-anti-MSLN in LS174T mice vs 3.27 ± 0.20 for 124I-anti-MSLN in HepG2 mice vs 3.53 ± 0.2 for 124I-IgG in LS174T mice). The immunochemical histology results showed that LS174T tumors were strongly positive (+++) for MSLN, while those in the HepG2 group showed slight expression (+). The dosimetry estimation study showed that the effective dose of 124I-anti-MSLN was 0.185 mSv/MBq, which is within the range of acceptable doses for further nuclear medicine translational research. Taken together, these results suggest that this radiotracer has the potential for detecting mesothelin-overexpressing tumors.
Assuntos
Neoplasias do Colo , Mesotelina , Animais , Linhagem Celular Tumoral , Proteínas Ligadas por GPI/metabolismo , Imunoglobulina G , Radioisótopos do Iodo , CamundongosRESUMO
Programmed death-1 (PD-1) and programmed death ligand-1 (PD-L1), the research focus in immune checkpoint regulation, play an important role in tumor immunotherapy. Inhibitors of this pathway are also the focus of tumor immunotherapy research. The PD-1/PD-L1 pathway can be blocked by selective binding to PD-L1. Clinical trials have been conducted in a variety of patients with advanced solid tumors. CS1001 is a high-affinity humanized full-length anti-PD-L1 monoclonal antibody with great clinical significance. We constructed a PD-L1-targeted radioactive molecular probe, 124/125I-labeled full-length antibody CS1001, and evaluated its binding specificity and targeting ability to PD-L1 in tumor cells and tumor models. Additionally, a comparison study with 68Ga-WL12, a PD-L1 targeting peptide, was conducted. The binding potency of 125I-CS1001 to human PD-L1 was evaluated by enzyme-linked immunosorbent assay (ELISA), and the Kd value was 52.1 ± 19.3 nM. The cellular uptake of 125I-CS1001 was examined in Chinese hamster ovary cells (CHO) and CHO expressing human PD-L1 (CHO-hPD-L1). At 2 h, the uptake values of 125I-CS1001 in CHO-hPD-L1 without blocking and in the presence of 0.1 mg non-radiolabeled CS1001 were 3.60 ± 0.08 and 0.09 ± 0.005 (%AD/2 × 105 cells, p < 0.001). Micro-PET imaging was performed between 8 to 192 h after injection of 124I-CS1001 into normal KM mice and CHO-hPD-L1 and HeLa tumor models. The standard uptake value (SUV) of relevant organs in PET images was calculated by drawing regions of interest (ROI). SUVmean of CHO-hPD-L1 tumors was significantly higher than that of HeLa tumors at 48 h (1.98 ± 0.04 vs 0.73 ± 0.14, p = 0.005). The SUVmean of 124I-CS1001 in CHO-hPD-L1 tumors at 48 h was higher than that of 68Ga-WL12 in CHO-hPD-L1 tumors at 0.5 h (1.98 ± 0.04 vs 1.09 ± 0.1 SUVmean, p = 0.007). In conclusion, this work provides a new method for monitoring and evaluating the in vivo expression of PD-L1 in tumors.
Assuntos
Iodo , Neoplasias , Cricetinae , Animais , Humanos , Camundongos , Células CHO , Radioisótopos de Gálio/química , Receptor de Morte Celular Programada 1 , Cricetulus , Anticorpos Monoclonais , Peptídeos/química , Tomografia por Emissão de Pósitrons/métodos , Linhagem Celular TumoralRESUMO
PEDOT: PSS/silicon heterojunction solar cell has recently attracted much attention due to the fact that it can be simply and cost-effectively fabricated. It is crucial to suppress the interfacial recombination rate between silicon (Si) and organic film for improving device efficiency. In this study, we demonstrated a thickness-dependent passivation effect, i.e. the passivation quality over Si substrate was promoted dramatically with increasing the thickness of PEDOT:PSS layer. The effective minority carrier lifetime increased from 32 µs for 50 nm to 360 µs for 200 nm, which corresponds to a change in implied open circuit voltage (V oc-implied) from 545 to 635 mV. Back-junction hybrid solar cells featuring PEDOT:PSS films at rear side were designed to enable adoption of thick PEDOT:PSS layers without having to worry about parasitic absorption, showing a power conversion efficiency (PCE) of 16.3%. Combined with a proper pre-condition on the Si substrate, the back-junction hybrid solar cell with 200 nm PEDOT:PSS layer received an enhanced PCE of 16.8%. In addition, the improved long-term stability for the back-junction device was also observed. The PCE remained 90% (unsealed) after being stored in ambient atmosphere for 30 days and over 80% (sealed) after 150 days.
RESUMO
Weakly supervised temporal action localization aims to locate the temporal boundaries of action instances in untrimmed videos using video-level labels and assign them the corresponding action category. Generally, it is solved by a pipeline called "localization-by-classification", which finds the action instances by classifying video snippets. However, since this approach optimizes the video-level classification objective, the generated activation sequences often suffer interference from class-related scenes, resulting in a large number of false positives in the prediction results. Many existing works treat background as an independent category, forcing models to learn to distinguish background snippets. However, under weakly supervised conditions, the background information is fuzzy and uncertain, making this method extremely difficult. To alleviate the impact of false positives, we propose a new actionness-guided false positive suppression framework. Our method seeks to suppress false positive backgrounds without introducing the background category. Firstly, we propose a self-training actionness branch to learn class-agnostic actionness, which can minimize the interference of class-related scene information by ignoring the video labels. Secondly, we propose a false positive suppression module to mine false positive snippets and suppress them. Finally, we introduce the foreground enhancement module, which guides the model to learn the foreground with the help of the attention mechanism as well as class-agnostic actionness. We conduct extensive experiments on three benchmarks (THUMOS14, ActivityNet1.2, and ActivityNet1.3). The results demonstrate the effectiveness of our method in suppressing false positives and it achieves the state-of-the-art performance. Code: https://github.com/lizhilin-ustc/AFPS.
Assuntos
Gravação em Vídeo , Humanos , Redes Neurais de Computação , Aprendizado de Máquina Supervisionado , AlgoritmosRESUMO
PURPOSE: Claudin 18.2 (CLDN18.2), due to its highly selective expression in tumor cells, has made breakthrough progress in clinical research and is expected to be integrated into routine tumor diagnosis and treatment. METHODS: In this research, we obtained an scFv-Fc fusion protein (SF106) targeting CLDN18.2 through hybridoma technology. The scFv-Fc fusion protein was labeled with radioactive isotopes (124I and 177Lu) to generate the radio-probes. The targeting and specificity of the radio-probes were tested in cellular models, and its diagnostic and therapeutic potential was further evaluated in tumor-bearing models. RESULTS: The molecular probes [124I]I-SF106 and [177Lu]Lu-DOTA-SF106 possess high radiochemical purity (RCP, 98.18 ± 0.93 % and 97.05 ± 1.1 %) and exhibit good stability in phosphate buffer saline and 5 % human serum albumin (92.44 ± 4.68 % and 91.03 ± 2.42 % at 120 h). [124I]I-SF106 uptake in cells expressing CLDN18.2 was well targeted and specific, and the dissociation constant was 17.74 nM [124I]I-SF106 micro-PET imaging showed that the maximum standardized uptake value (SUVmax) was significantly higher than CLDN18.2-negative tumors (1.83 ± 0.02 vs. 1.23 ± 0.04, p < 0.001). The maximum uptake was attained in tumors expressing CLDN18.2 at 48 h after injection. [124I]I-SF106 and [177Lu]Lu-DOTA-SF106 dosimetric study showed that the effective dose in humans complies with the medical safety standards required for their clinical application. The results of treatment experiments showed that 3 MBq of [177Lu]Lu-DOTA-SF106 in CLDN18.2-expressing tumor-bearing mice could significantly inhibit tumor growth. CONCLUSION: These results indicate that radionuclide-labeled scFv-Fc molecular probes ([124I]I-SF106 and [177Lu]Lu-DOTA-SF106) provide a new possibility for the diagnosis and treatment of CLDN18.2-positive cancer patients in clinical practice.
Assuntos
Neoplasias , Compostos Radiofarmacêuticos , Humanos , Camundongos , Animais , Compostos Radiofarmacêuticos/farmacologia , Compostos Radiofarmacêuticos/uso terapêutico , Neoplasias/diagnóstico por imagem , Neoplasias/terapia , Albumina Sérica Humana , Radioisótopos do Iodo , Sondas Moleculares , Linhagem Celular Tumoral , ClaudinasRESUMO
BACKGROUND: The lack of an efficient way to screen patients who are responsive to immunotherapy challenges PD1/CTLA4-targeting cancer treatment. Immunotherapeutic efficacy cannot be clearly determined by peripheral blood analyses, tissue gene markers or CT/MR value. Here, we used a radionuclide and imaging techniques to investigate the novel dual targeted antibody cadonilimab (AK104) in PD1/CTLA4-positive cells in vivo. METHODS: First, humanized PD1/CTLA4 mice were purchased from Biocytogen Pharmaceuticals (Beijing) Co., Ltd. to express hPD1/CTLA4 in T-cells. Then, mouse colon cancer MC38-hPD-L1 cell xenografts were established in humanized mice. A bispecific antibody targeting PD1/CTLA4 (AK104) was labeled with radio-nuclide iodine isotopes. Immuno-PET/CT imaging was performed using a bispecific monoclonal antibody (mAb) probe 124I-AK104, developed in-house, to locate PD1+/CTLA4+ tumor-infiltrating T cells and monitor their distribution in mice to evaluate the therapeutic effect. RESULTS: The 124I-AK104 dual-antibody was successfully constructed with ideal radiochemical characteristics, in vitro stability and specificity. The results of immuno-PET showed that 124I-AK104 revealed strong hPD1/CTLA4-positive responses with high specificity in humanized mice. High uptake of 124I-AK104 was observed not only at the tumor site but also in the spleen. Compared with PD1- or CTLA4-targeting mAb imaging, 124I-AK104 imaging had excellent standard uptake values at the tumor site and higher tumor to nontumor (T/NT) ratios. CONCLUSIONS: The results demonstrated the potential of translating 124I-AK104 into a method for screening patients who benefit from immunotherapy and the efficacy, as well as the feasibility, of this method was verified by immuno-PET imaging of humanized mice.
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Anticorpos Biespecíficos , Antígeno CTLA-4 , Tomografia por Emissão de Pósitrons combinada à Tomografia Computadorizada , Receptor de Morte Celular Programada 1 , Animais , Anticorpos Biespecíficos/farmacologia , Anticorpos Biespecíficos/imunologia , Humanos , Camundongos , Antígeno CTLA-4/imunologia , Linhagem Celular Tumoral , Tomografia por Emissão de Pósitrons combinada à Tomografia Computadorizada/métodos , Receptor de Morte Celular Programada 1/imunologia , Neoplasias do Colo/diagnóstico por imagem , Neoplasias do Colo/imunologia , Neoplasias do Colo/tratamento farmacológico , Neoplasias do Colo/patologia , Radioisótopos do Iodo , Ensaios Antitumorais Modelo de Xenoenxerto , Distribuição Tecidual , Linfócitos do Interstício Tumoral/imunologia , Linfócitos do Interstício Tumoral/efeitos dos fármacos , FemininoRESUMO
Angiotensin-converting enzyme 2 (ACE2) is the main molecular target for coronavirus SARS-CoV-2 to enter cells. Molecularly specific tracers that bind to ACE2 with high affinity can be used to determine the tissue distribution of this important receptor, noninvasively. A novel targeting PET imaging probe, [18F]AlF-DX600-BCH, was developed to detect the in vivo expression of ACE2 and monitor response to therapy. Preclinical experiments, including biodistribution, PET imaging, and tissue section analysis, were conducted after tests of in vitro and in vivo stability and pharmacokinetics. The agent was advanced to clinical evaluation in 10 volunteers who received [18F]AlF-DX600-BCH PET/CT at 1 and 2 h after injection (NCT04542863). Preclinical results of both biodistribution and PET demonstrated [18F]AlF-DX600-BCH accumulation in rat kidney (standardized uptake value; SUVkidney/normal > 50), along with specific uptake in testes (SUVtestis/normal > 10) tissues. Kidney, gastrointestinal, and bronchial cell labeling were correlated to ACE2 positive by immunohistochemistry (IHC) staining. In clinical imaging, significant tracer accumulation was predominantly observed in the urinary and reproductive system (SUVrenal cortex = 32.00, SUVtestis = 4.56), and the conjunctiva and nasal mucosa saw elevated uptake in several cases. This work is the first report of a radioisotope probe, [18F]AlF-DX600-BCH, targeting ACE2 with promising preliminary preclinical and translational outlook, thereby demonstrating the potential of noninvasive mapping of ACE2.
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Recent global clinical trials have shown that CLDN18.2 is an ideal target for the treatment of gastric cancer and that patients with high CLDN18.2 expression can benefit from targeted therapy. Therefore, accurate and comprehensive detection of CLDN18.2 expression is important for patient screening and guidance in anti-CLDN18.2 therapy. Phage display technology was used to screen CLDN18.2-specific peptides from 100 billion libraries. 293TCLDN18.1 cells were used to exclude nonspecific binding and CLDN18.1 binding sequences, while 293TCLDN18.2 cells were used to screen CLDN18.2-specific binding peptides. The monoclonal clones obtained from phage screening were sequenced, and peptides were synthesized based on the sequencing results. Binding specificity and affinity were assessed with a fluorescein isothiocyanate (FITC)-conjugated peptide. A 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid (DOTA)-conjugated peptide was also synthesized for 68Ga radiolabeling. The in vitro and in vivo stability, partition coefficients, in vivo molecular imaging, and biodistribution were also characterized. Overall, 54 monoclonal clones were selected after phage display screening. Subsequently, based on the cell ELISA results, CLDN18.2 preference monoclonal clones were selected for deoxyribonucleic acid (DNA) sequencing, and four 7-peptide sequences were obtained after sequence comparison; among them, a peptide named T37 was further validated in vitro and in vivo. The T37 peptide specifically recognized CLDN18.2 but not CLDN18.1 and bound strongly to CLDN18.2-positive cell membranes. The 68Ga-DOTA-T37 probe exhibits good in vitro properties and high stability as a hydrophilic probe; it has high biological safety, and positron emission tomography/computed tomography (PET/CT) studies have shown that it can specifically target CLDN18.2 protein and CLDN18.2-positive tumors in mice. 68Ga-DOTA-T37 demonstrated the superiority and feasibility of using a CLDN18.2-specific probe in PCT/CT imaging, which deserves further development and exploitation.
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Claudin18.2 (CLDN18.2) is a tight junction protein that is overexpressed in a variety of solid tumors such as gastrointestinal cancer and oesophageal cancer. It has been identified as a promising target and a potential biomarker to diagnose tumor, evaluate efficacy, and determine patient prognosis. TST001 is a recombinant humanized CLDN18.2 antibody that selectively binds to the extracellular loop of human Claudin18.2. In this study, we constructed a solid target radionuclide zirconium-89 (89Zr) labled-TST001 to detect the expression of in the human stomach cancer BGC823CLDN18.2 cell lines. The [89Zr]Zr-desferrioxamine (DFO)-TST001 showed high radiochemical purity (RCP, >99%) and specific activity (24.15 ± 1.34 GBq/µmol), and was stable in 5% human serum albumin, and phosphate buffer saline (>85% RCP at 96 h). The EC50 values of TST001 and DFO-TST001 were as high as 0.413 ± 0.055 and 0.361 ± 0.058 nM (P > 0.05), respectively. The radiotracer had a significantly higher average standard uptake values in CLDN18.2-positive tumors than in CLDN18.2-negative tumors (1.11 ± 0.02 vs. 0.49 ± 0.03, P = 0.0016) 2 days post injection (p.i.). BGC823CLDN18.2 mice models showed high tumor/muscle ratios 96 h p.i. with [89Zr]Zr-DFO-TST001 was much higher than those of the other imaging groups. Immunohistochemistry results showed that BGC823CLDN18.2 tumors were highly positive (+++) for CLDN18.2, while those in the BGC823 group did not express CLDN18.2 (-). The results of ex vivo biodistribution studies showed that there was a higher distribution in the BGC823CLDN18.2 tumor bearing mice (2.05 ± 0.16 %ID/g) than BGC823 mice (0.69 ± 0.02 %ID/g) and blocking group (0.72 ± 0.02 %ID/g). A dosimetry estimation study showed that the effective dose of [89Zr]Zr-DFO-TST001 was 0.0705 mSv/MBq, which is within the range of acceptable doses for nuclear medicine research. Taken together, these results suggest that Good Manufacturing Practices produced by this immuno-positron emission tomography probe can detect CLDN18.2-overexpressing tumors.
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Traditional convolutional neural networks (CNNs) share their kernels among all positions of the input, which may constrain the representation ability in feature extraction. Dynamic convolution proposes to generate different kernels for different inputs to improve the model capacity. However, the total parameters of the dynamic network can be significantly huge. In this article, we propose a lightweight dynamic convolution method to strengthen traditional CNNs with an affordable increase of total parameters and multiply-adds. Instead of generating the whole kernels directly or combining several static kernels, we choose to "look inside", learning the attention within convolutional kernels. An extra network is used to adjust the weights of kernels for every feature aggregation operation. By combining local and global contexts, the proposed approach can capture the variance among different samples, the variance in different positions of the feature maps, and the variance in different positions inside sliding windows. With a minor increase in the number of model parameters, remarkable improvements in image classification on CIFAR and ImageNet with multiple backbones have been obtained. Experiments on object detection also verify the effectiveness of the proposed method.
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The ACE2 receptor, as the potential entrance site of SARS-CoV-2-affected cells, plays a crucial role in spreading infection. The DX600 peptide is a competitive inhibitor of ACE2. We previously constructed the 68Ga-labeled DOTA-DX600 (also known as 68Ga-HZ20) peptide and confirmed its ACE2 binding ability both in vitro and in vivo. In this research, we aimed to investigate the noninvasive mapping of ACE2 expression in fowl using 68Ga-HZ20 micro-PET. We chose pigeons as an animal model and first studied the administration method of 68Ga-HZ20 by direct site injection or intravenous injection. Then, the dynamic micro-PET scan of 68Ga-HZ20 was conducted at 0-40 min. Additionally, 18F-FDG was used for comparison. Finally, the pigeons were sacrificed, and the main organs were collected for further immunoPET and IHC staining. Micro PET/CT imaging results showed that 68Ga-HZ20 uptake was distributed from the heart at the preliminary injection to the kidneys, liver, stomach, and lungs over time, where the highest uptake was observed in the kidneys (SUVmax = 6.95, 20 min) and lung (SUVmax = 1.11, 20 min). Immunohistochemical experiments were carried out on its main organs. Compared to the SUVmax data, the IHC results showed that ACE2 was highly expressed in both kidneys and intestines, and the optimal imaging time was determined to be 20 min after injection through correlation analysis. These results indicated that 68Ga-HZ20 is a potential target molecule for SARS-CoV-2 in fowl, which is worthy of promotion and further study.
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PURPOSE: To develop and test semi-supervised generative adversarial networks (GANs) that detect retinal disorders on optical coherence tomography (OCT) images using a small-labeled dataset. METHODS: From a public database, we randomly chose a small supervised dataset with 400 OCT images (100 choroidal neovascularization, 100 diabetic macular edema, 100 drusen, and 100 normal) and assigned all other OCT images to unsupervised dataset (107,912 images without labeling). We adopted a semi-supervised GAN and a supervised deep learning (DL) model for automatically detecting retinal disorders from OCT images. The performance of the 2 models was compared in 3 testing datasets with different OCT devices. The evaluation metrics included accuracy, sensitivity, specificity, and the area under the receiver operating characteristic curves. RESULTS: The local validation dataset included 1000 images with 250 from each category. The independent clinical dataset included 366 OCT images using Cirrus OCT Shanghai Shibei Hospital and 511 OCT images using RTVue OCT from Xinhua Hospital respectively. The semi-supervised GANs classifier achieved better accuracy than supervised DL model (0.91 vs 0.86 for local cell validation dataset, 0.91 vs 0.86 in the Shanghai Shibei Hospital testing dataset, and 0.93 vs 0.92 in Xinhua Hospital testing dataset). For detecting urgent referrals (choroidal neo-vascularization and diabetic macular edema) from nonurgent referrals (drusen and normal) on OCT images, the semi-supervised GANs classifier also achieved better area under the receiver operating characteristic curves than supervised DL model (0.99 vs 0.97, 0.97 vs 0.96, and 0.99 vs 0.99, respectively). CONCLUSIONS: A semi-supervised GAN can achieve better performance than that of a supervised DL model when the labeled dataset is limited. The current study offers utility to various research and clinical studies using DL with relatively small datasets. Semi-supervised GANs can detect retinal disorders from OCT images using relatively small dataset.
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Retinopatia Diabética , Edema Macular , Doenças Retinianas , Tomografia de Coerência Óptica , Algoritmos , China , Aprendizado Profundo , Retinopatia Diabética/diagnóstico por imagem , Humanos , Edema Macular/diagnóstico por imagem , Doenças Retinianas/diagnóstico por imagem , Aprendizado de Máquina Supervisionado , Tomografia de Coerência Óptica/métodosRESUMO
Combining electron- and hole-selective materials in one crystalline silicon (Si) solar cell, thereby avoiding any dopants, is not considered for application to photovoltaic industry until only comparable efficiency and stable performance are achievable. Here, it is demonstrated how a conventionally unstable electron-selective contact (ESC) is optimized with huge boost in stability as well as improved electron transport. With the introduction of a Ti thin film between a-Si:H(i)/LiF and Al electrode, high-level passivation (Seff = 4.6 cm s-1 ) from a-Si:H(i) and preferential band alignment (ρC = 7.9 mΩ cm2 ) from low work function stack of LiF/Ti/Al are both stably retained in the newly constructed n-Si/a-Si:H(i)/LiF/Ti/Al ESC. A detailed interfacial elements analysis reveals that the efficiently blocked inward diffusion of Al from electrode by the Ti protecting layer balances transport and recombination losses in general. This excellent electron-selective properties in combination with large process tolerance that enable remarkable device performance, particularly high efficiencies of 22.12% and 23.61%, respectively, are successfully approached by heterojunction solar cells with dopant-free ESC and dopant-free contacts for both polarities.