Seokgyu Han, Ingyoung Kim, Baekcheon Seong, Woovin Kim, Hongseong Kim, Sein Kim, Chulmin Joo, Sungsu Park
{"title":"衰减和后向散射系数的双参数层析成像定量评价免疫细胞介导的肿瘤球体细胞毒性。","authors":"Seokgyu Han, Ingyoung Kim, Baekcheon Seong, Woovin Kim, Hongseong Kim, Sein Kim, Chulmin Joo, Sungsu Park","doi":"10.7150/thno.118722","DOIUrl":null,"url":null,"abstract":"<p><p><b>Rationale:</b> Quantitative, non-perturbative assessment of immune cell-mediated cytotoxicity in tumor spheroids remains challenging due to the lack of real-time label-free analytical tools. Conventional methods such as fluorescence imaging or biochemical assays often require labeling and provide limited longitudinal analysis, which prohibits dynamic monitoring of therapeutic responses. This study presents a dual-parameter tomographic analysis method that simultaneously quantifies attenuation coefficient (AC) and backscattering coefficient (BSC) from optical coherence tomography (OCT) datasets, enabling dynamic evaluation of therapeutic responses in three-dimensional (3D) tumor spheroids. <b>Methods:</b> We developed a 3D Gabor transform-based algorithm to extract depth-resolved AC and BSC metrics from OCT volumetric datasets. Unlike conventional strategies, our method enables simultaneous voxel-wise measurements of AC and BSC values, with superior noise robustness. Experiments with intralipid solutions across a range of concentrations revealed that the Gabor-based approach yielded AC and BSC estimations with more than three times greater precision than prior methods. This approach enables high-resolution measurements of structural and optical property changes associated with apoptosis, allowing spatial and temporal mapping of treatment-induced cytotoxicity in HER2-positive breast tumor spheroids treated with AZD4547 and HER2-targeted chimeric antigen receptor (CAR) T cells. <b>Results:</b> In AZD4547-treated spheroids, AC increased dose-dependently from 0.39 to 0.64, reflecting a 64% rise, while BSC increased from 0.09 to 0.12, an approximate 33% increase. CAR T cell treatment induced a rapid, spatially progressive increase in both AC and BSC, originating at the spheroid periphery and advancing inward. Over 12 hours, AC rose from 0.40 to 0.82 (2-fold increase) and BSC from 0.09 to 0.20 (2.2-fold increase). While AC and BSC individually correlated with spheroid viability, their combined analysis consistently achieved a higher coefficient of determination (R² = 0.98) across both treatment modalities. <b>Conclusions:</b> This dual-parameter OCT-based assay framework provides a sensitive, label-free method for distinguishing between immune- and drug-induced apoptosis in tumor spheroids. Its strong correlation with viability and capacity to resolve spatially resolved dynamics underscore its potential for robust, <i>in situ</i> assessment of immunotherapeutic efficacy.</p>","PeriodicalId":22932,"journal":{"name":"Theranostics","volume":"15 18","pages":"9399-9414"},"PeriodicalIF":13.3000,"publicationDate":"2025-08-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12486019/pdf/","citationCount":"0","resultStr":"{\"title\":\"Dual-parameter tomographic imaging of attenuation and backscattering coefficients for quantitative evaluation of immune cell-mediated cytotoxicity in tumor spheroids.\",\"authors\":\"Seokgyu Han, Ingyoung Kim, Baekcheon Seong, Woovin Kim, Hongseong Kim, Sein Kim, Chulmin Joo, Sungsu Park\",\"doi\":\"10.7150/thno.118722\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p><b>Rationale:</b> Quantitative, non-perturbative assessment of immune cell-mediated cytotoxicity in tumor spheroids remains challenging due to the lack of real-time label-free analytical tools. Conventional methods such as fluorescence imaging or biochemical assays often require labeling and provide limited longitudinal analysis, which prohibits dynamic monitoring of therapeutic responses. This study presents a dual-parameter tomographic analysis method that simultaneously quantifies attenuation coefficient (AC) and backscattering coefficient (BSC) from optical coherence tomography (OCT) datasets, enabling dynamic evaluation of therapeutic responses in three-dimensional (3D) tumor spheroids. <b>Methods:</b> We developed a 3D Gabor transform-based algorithm to extract depth-resolved AC and BSC metrics from OCT volumetric datasets. Unlike conventional strategies, our method enables simultaneous voxel-wise measurements of AC and BSC values, with superior noise robustness. Experiments with intralipid solutions across a range of concentrations revealed that the Gabor-based approach yielded AC and BSC estimations with more than three times greater precision than prior methods. This approach enables high-resolution measurements of structural and optical property changes associated with apoptosis, allowing spatial and temporal mapping of treatment-induced cytotoxicity in HER2-positive breast tumor spheroids treated with AZD4547 and HER2-targeted chimeric antigen receptor (CAR) T cells. <b>Results:</b> In AZD4547-treated spheroids, AC increased dose-dependently from 0.39 to 0.64, reflecting a 64% rise, while BSC increased from 0.09 to 0.12, an approximate 33% increase. CAR T cell treatment induced a rapid, spatially progressive increase in both AC and BSC, originating at the spheroid periphery and advancing inward. Over 12 hours, AC rose from 0.40 to 0.82 (2-fold increase) and BSC from 0.09 to 0.20 (2.2-fold increase). While AC and BSC individually correlated with spheroid viability, their combined analysis consistently achieved a higher coefficient of determination (R² = 0.98) across both treatment modalities. <b>Conclusions:</b> This dual-parameter OCT-based assay framework provides a sensitive, label-free method for distinguishing between immune- and drug-induced apoptosis in tumor spheroids. Its strong correlation with viability and capacity to resolve spatially resolved dynamics underscore its potential for robust, <i>in situ</i> assessment of immunotherapeutic efficacy.</p>\",\"PeriodicalId\":22932,\"journal\":{\"name\":\"Theranostics\",\"volume\":\"15 18\",\"pages\":\"9399-9414\"},\"PeriodicalIF\":13.3000,\"publicationDate\":\"2025-08-22\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12486019/pdf/\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Theranostics\",\"FirstCategoryId\":\"3\",\"ListUrlMain\":\"https://doi.org/10.7150/thno.118722\",\"RegionNum\":1,\"RegionCategory\":\"医学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"2025/1/1 0:00:00\",\"PubModel\":\"eCollection\",\"JCR\":\"Q1\",\"JCRName\":\"MEDICINE, RESEARCH & EXPERIMENTAL\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Theranostics","FirstCategoryId":"3","ListUrlMain":"https://doi.org/10.7150/thno.118722","RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2025/1/1 0:00:00","PubModel":"eCollection","JCR":"Q1","JCRName":"MEDICINE, RESEARCH & EXPERIMENTAL","Score":null,"Total":0}
Dual-parameter tomographic imaging of attenuation and backscattering coefficients for quantitative evaluation of immune cell-mediated cytotoxicity in tumor spheroids.
Rationale: Quantitative, non-perturbative assessment of immune cell-mediated cytotoxicity in tumor spheroids remains challenging due to the lack of real-time label-free analytical tools. Conventional methods such as fluorescence imaging or biochemical assays often require labeling and provide limited longitudinal analysis, which prohibits dynamic monitoring of therapeutic responses. This study presents a dual-parameter tomographic analysis method that simultaneously quantifies attenuation coefficient (AC) and backscattering coefficient (BSC) from optical coherence tomography (OCT) datasets, enabling dynamic evaluation of therapeutic responses in three-dimensional (3D) tumor spheroids. Methods: We developed a 3D Gabor transform-based algorithm to extract depth-resolved AC and BSC metrics from OCT volumetric datasets. Unlike conventional strategies, our method enables simultaneous voxel-wise measurements of AC and BSC values, with superior noise robustness. Experiments with intralipid solutions across a range of concentrations revealed that the Gabor-based approach yielded AC and BSC estimations with more than three times greater precision than prior methods. This approach enables high-resolution measurements of structural and optical property changes associated with apoptosis, allowing spatial and temporal mapping of treatment-induced cytotoxicity in HER2-positive breast tumor spheroids treated with AZD4547 and HER2-targeted chimeric antigen receptor (CAR) T cells. Results: In AZD4547-treated spheroids, AC increased dose-dependently from 0.39 to 0.64, reflecting a 64% rise, while BSC increased from 0.09 to 0.12, an approximate 33% increase. CAR T cell treatment induced a rapid, spatially progressive increase in both AC and BSC, originating at the spheroid periphery and advancing inward. Over 12 hours, AC rose from 0.40 to 0.82 (2-fold increase) and BSC from 0.09 to 0.20 (2.2-fold increase). While AC and BSC individually correlated with spheroid viability, their combined analysis consistently achieved a higher coefficient of determination (R² = 0.98) across both treatment modalities. Conclusions: This dual-parameter OCT-based assay framework provides a sensitive, label-free method for distinguishing between immune- and drug-induced apoptosis in tumor spheroids. Its strong correlation with viability and capacity to resolve spatially resolved dynamics underscore its potential for robust, in situ assessment of immunotherapeutic efficacy.
期刊介绍:
Theranostics serves as a pivotal platform for the exchange of clinical and scientific insights within the diagnostic and therapeutic molecular and nanomedicine community, along with allied professions engaged in integrating molecular imaging and therapy. As a multidisciplinary journal, Theranostics showcases innovative research articles spanning fields such as in vitro diagnostics and prognostics, in vivo molecular imaging, molecular therapeutics, image-guided therapy, biosensor technology, nanobiosensors, bioelectronics, system biology, translational medicine, point-of-care applications, and personalized medicine. Encouraging a broad spectrum of biomedical research with potential theranostic applications, the journal rigorously peer-reviews primary research, alongside publishing reviews, news, and commentary that aim to bridge the gap between the laboratory, clinic, and biotechnology industries.