Laboratory Investigation最新文献_第8页

Multiscale Fusion Models With Genomic, Topological, and Pathomic Features to Predict Response to Radiation Therapy for Non–Small Cell Lung Cancer Patients 具有基因组、拓扑和病理特征的多尺度融合模型预测非小细胞肺癌患者对放射治疗的反应。

IF 5.1 2区医学 Q1 MEDICINE, RESEARCH & EXPERIMENTAL

Laboratory Investigation

Pub Date : 2025-06-16 DOI: 10.1016/j.labinv.2025.104204

Yu Jin , Hidetaka Arimura , Takeshi Iwasaki , Takumi Kodama , Noriaki Yamamoto , Yunhao Cui , Yoshinao Oda

Artificial intelligence models with biomarkers to predict treatment responses to radiation would be necessary to maximize the treatment outcomes of individual patients, especially with histopathology images routinely obtained before treatment. We hypothesized that multiscale features, such as genomic (GM), pathomic (PM), and topological (TP) features, could be associated with the radiation response. We investigated fusion models with multiscale features in histopathology images to predict response to radiation therapy for patients (responders) with non–small cell lung cancer. Ten radiosensitivity-related (radiosensitive and radioresistant) genes were deployed as GM features. PM features were extracted from histopathology images by conventional PM analyses. TP features represent the intrinsic properties of tumor cells using Betti numbers, which are mathematical invariants. We analyzed non–small cell lung cancer patients from The Cancer Genome Atlas and Clinical Proteomic Tumor Analysis Consortium who received radiotherapy and established 3 base models with GM, TP, and PM features, respectively, and 3 fusion models. The TP model showed a higher area under the receiver operating characteristic curve of 0.707 (P = .026, log-rank test in overall survival analysis) in the internal test data set and 0.720 (P = .136) in the external test data set. The results indicated that the TP models achieved better classification and prognostic prediction powers than the other base models. The inner-cell TP structure may have the ability to reveal the cell radiosensitivity-related information. Furthermore, the best fusion model with GM, TP, and PM features achieved the highest area under the receiver operating characteristic curve of 0.846 (P = .019) and 0.731 (P = .043) in predicting the treatment response and prognoses in the internal and external test data sets, respectively. This study demonstrated the predictive power of the multiscale fusion model for histopathology images, which may assist clinical physicians in the selection of responders to radiation for personalized radiation therapy and would be substantially beneficial for patients with cancer.

为了最大限度地提高个体患者的治疗效果，特别是治疗前常规获得的组织病理学图像，有必要使用生物标志物来预测治疗对辐射的反应的人工智能模型。我们假设多尺度特征，如基因组、病理和拓扑特征，可能与辐射反应有关。我们研究了组织病理学图像中具有多尺度特征的融合模型，以预测非小细胞肺癌患者（响应者）对放射治疗的反应。10个放射敏感性相关（放射敏感和放射抗性）基因被部署为基因组特征。通过常规病理分析从组织病理图像中提取病理特征。拓扑特征用数学不变量贝蒂数表示肿瘤细胞的内在特性。我们分析了TCGA和CPTAC接受放疗的非小细胞肺癌（NSCLC）患者，并分别建立了具有基因组、拓扑和病理特征的三种基础模型和三种融合模型。拓扑模型显示，内部测试数据集的受试者工作特征曲线（AUC）下面积为0.707 （p值=0.026，总生存分析中log-rank检验），外部测试数据集的受试者工作特征曲线下面积为0.720 （p值=0.136）。结果表明，拓扑模型比其他基础模型具有更好的分类和预测能力。细胞内拓扑结构可能具有揭示细胞辐射敏感性相关信息的能力。此外，在预测内部和外部测试数据集的治疗反应和预后时，具有基因组、拓扑和病理特征的最佳融合模型分别达到0.846 （p值=0.019）和0.731 （p值=0.043）的最高AUC。本研究证明了组织病理学图像的多尺度融合模型的预测能力，它可以帮助临床医生选择对个性化放射治疗有反应的放射患者，并且对癌症患者非常有益。

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引用次数: 0

Corrigendum to “386 Targeted NGS for TP53 and BRCA1, BRCA2, Gene Mutations and Comparison with Immune Cell Profiling and Immune Checkpoint Inhibitor Expression in Ascitic Fluid with Metastases from High Grade Serous Ovarian Carcinoma” [Laboratory Investigation 105 (2025) 102613] “386靶向NGS治疗高级别浆液性卵巢癌转移的腹水中TP53和BRCA1、BRCA2基因突变及免疫细胞谱和免疫检查点抑制剂表达的比较”的更正[实验室调查105 (2025)102613]

IF 5.1 2区医学 Q1 MEDICINE, RESEARCH & EXPERIMENTAL

Laboratory Investigation

Pub Date : 2025-06-13 DOI: 10.1016/j.labinv.2025.104173

Radhika Srinivasan , Megha Sharma , Amit Sharma , Bhavneet Kaur , Bhavana Rai , Man Updesh Sachdeva , Parikshaa Gupta , Upasana Gautam , Rashmi Bagga

引用次数: 0

Siglec Ligand Immunohistochemistry Reveals Association With Immune Exclusion and Survival Siglec配体免疫组化显示与免疫排斥和生存有关。

IF 5.1 2区医学 Q1 MEDICINE, RESEARCH & EXPERIMENTAL

Laboratory Investigation

Pub Date : 2025-06-09 DOI: 10.1016/j.labinv.2025.104203

Jeremy R. Ellis , Elizabeth Will , Aleksandra Ogurtsova , Logan L. Engle , Janis M. Taube , Joel C. Sunshine

Sialic acids are overexpressed in many cancers, and binding of sialic acid via sialic acid binding immunoglobulin-like lectins (Siglecs) may contribute significantly to immune evasion and cancer progression. This important resistance mechanism in the tumor immune microenvironment has been understudied, partially due to the lack of useful reagents. Here, we developed and optimized an immunohistochemistry staining protocol for novel reagents that detect 3 types of Siglec-engaging sialoglycans (HYDRA-3, -7, and -9, which detect sialoglycans recognized by Siglec-3, -7, and -9, respectively) in the tumor immune microenvironment. We evaluated HYDRA staining across 10 different cancer types across whole slides, finding that HYDRA-9 exhibited the highest overall staining range, with HYDRA-3 and -7 showing lower to moderate staining across all tested tumor types. To correlate HYDRA staining patterns and immune infiltration in melanoma, we stained melanoma tissue microarrays with the 3 HYDRA reagents and compared HYDRA staining profiles with a 6-plex multiplex immunofluorescence panel targeting CD8, CD163, FoxP3, PD1, PDL1, and Sox10/S100. Siglec-3 and -9 sialoglycan ligand expression negatively correlated with CD8 T cell infiltration (r = −0.28/P = .002 and r = −0.29/P = .001, respectively), particularly at the tumor-stromal interface (r = −0.37/P < .001 and r = −0.44/P < .001, respectively). Additionally, a high ratio of Siglec-3 and -9 ligand expression at the tumor-stromal interface versus the tumor core was associated with reduced overall survival (Hazard’s ratio: 2.60 and 2.11, respectively), whereas CD8 infiltration was not associated with survival outcomes in our cohort. Taken together, the expression levels and spatial distribution of Siglec-engaging sialoglycans may play a role in patient prognosis, potentially representing a biomarker of survival that is independent of conventional metrics of an inflamed tumor microenvironment. This study highlights the need for further investigation of Siglec ligand expression as a predictive and prognostic biomarker of treatment response and resistance.

唾液酸在许多癌症中过度表达，唾液酸通过唾液酸结合免疫球蛋白样凝集素（Siglecs）结合可能对免疫逃避和癌症进展起重要作用。肿瘤免疫微环境（TIME）中这种重要的耐药机制尚未得到充分研究，部分原因是缺乏有用的试剂。在这里，我们开发并优化了一种免疫组织化学（IHC）染色方案，用于检测三种类型的siglece唾液聚糖（HYDRA-3， -7和-9，分别检测siglec3， -7和-9识别的唾液聚糖）。我们在整个载玻片上评估了HYDRA在10种不同癌症类型中的染色情况，发现HYDRA-9在所有测试的肿瘤类型中表现出最高的总体染色范围，而HYDRA-3和-7在所有测试的肿瘤类型中表现出较低到中等的染色。为了将HYDRA染色模式与黑色素瘤中的免疫浸润联系起来，我们用3种HYDRA试剂对黑色素瘤组织微阵列（tma）进行染色，并将HYDRA染色谱与靶向CD8、CD163、FoxP3、PD1、PDL1和Sox10/S100的6-plex多重免疫荧光面板进行比较。Siglec-3和-9唾液聚糖配体表达与CD8 T细胞浸润呈负相关（r=-0.28/p=0.002和r=-0.29/p=0.001），特别是在肿瘤-基质界面（r= -0.37/p）

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引用次数: 0

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IF 5.1 2区医学 Q1 MEDICINE, RESEARCH & EXPERIMENTAL

Laboratory Investigation

Pub Date : 2025-06-01 DOI: 10.1016/S0023-6837(25)00104-7

引用次数: 0

Histopathological Insights into Metastatic Breast Cancer Gained From Rapid Autopsies 从快速尸检中获得转移性乳腺癌的组织病理学见解。

IF 5.1 2区医学 Q1 MEDICINE, RESEARCH & EXPERIMENTAL

Laboratory Investigation

Pub Date : 2025-05-29 DOI: 10.1016/j.labinv.2025.104202

Gitte Zels , Anirudh Pabba , Maxim De Schepper , Tatjana Geukens , Karen Van Baelen , Marion Maetens , Amena Mahdami , Sophia Leduc , Edoardo Isnaldi , Ha-Linh Nguyen , Imane Bachir , Josephine Van Cauwenberge , Kristien Borremans , Birgit Weynand , Elia Biganzoli , Patrick Neven , Hans Wildiers , Wouter Van Den Bogaert , François Richard , Christine Desmedt , Giuseppe Floris

The biology of metastatic breast cancer is poorly understood, and its understanding is hampered by limited access to metastatic tissue. Post-mortem tissue donation programs may represent a step forward to circumvent this problem, allowing access to large volumes of samples that would often be inaccessible otherwise. In this context, we have set up the UZ/KU Leuven Program for post-mortem Tissue Donation to Enhance Research (UPTIDER, NCT04531696). In this study, we performed detailed histopathological examination of 662 unique metastases collected during autopsy from the first 20 patients in our UPTIDER program. Tissue procurement was guided by a patient-specific tissue donation plan based on available clinical and imaging data. Central pathology review included revision of the primary tumor and biomarker assessment (estrogen receptor [ER], progesterone receptor [PR], human epidermal growth factor receptor 2, and KI67). Linear mixed quantile regression was used to assess relevant associations. Metastases in bones, liver, pleura, and nonaxillary lymph nodes were present in up to 17 patients. Our major findings include (1) an important clinical underestimation of disease burden in patients with invasive lobular carcinoma; (2) a relatively modest disease burden associated with leptomeningeal metastases; (3) a higher than anticipated loss of predictive biomarkers in metastases from primary ER- positive and/or PR positive tumors (up to 84% and 100% of the patients having at least 1 ER-negative or PR-negative lesion, respectively); (4) a high variability in KI67% between metastases with frequent zonation pattern; and (5) a high frequency of metastases with human epidermal growth factor receptor 2–low or –ultralow status. Despite the challenging setup of UPTIDER, we demonstrated here that the data and observations that emerge from this program have high potential for clinical translatability.

对转移性乳腺癌的生物学了解甚少，而且由于转移组织的接触有限，对转移性乳腺癌的了解也受到阻碍。死后组织捐赠项目可能是解决这一问题的一个进步，它使人们能够获得大量的样本，否则这些样本通常是无法获得的。在此背景下，我们建立了鲁汶大学/鲁汶大学尸体组织捐赠计划，以加强研究（uptid, NCT04531696）。在这项研究中，我们对在UPTIDER项目的前20名患者的尸检中收集的662例独特转移瘤进行了详细的组织病理学检查。组织采购由基于现有临床和影像学数据的患者特异性组织捐赠计划指导。中心病理检查包括原发肿瘤的修订和生物标志物评估（ER、PR、HER2和KI67）。采用线性混合分位数回归评估相关关联。17例患者出现骨、肝、胸膜和非腋窝淋巴结转移。我们的主要发现包括：a)侵袭性小叶癌患者疾病负担的重要临床低估；B)与脑膜轻脑膜转移相关的相对适度的疾病负担；c)原发性ER+和/或PR+肿瘤转移灶中预测性生物标志物的损失高于预期（分别高达84%和100%的患者至少有一个ER阴性或PR阴性病变），d) KI67%的高变异性，具有频繁的分区模式，e)转移灶的her2低或-超低状态的频率很高。尽管UPTIDER的设置具有挑战性，但我们在这里证明，从该程序中出现的数据和观察结果具有很高的临床可翻译性潜力。

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引用次数: 0

Integrative Genetic and Transcriptomic Subtyping Improves Prognosis Prediction in B-Lineage Acute Lymphoblastic Leukemia 综合遗传和转录组亚分可改善b系急性淋巴细胞白血病的预后预测。

IF 5.1 2区医学 Q1 MEDICINE, RESEARCH & EXPERIMENTAL

Laboratory Investigation

Pub Date : 2025-05-29 DOI: 10.1016/j.labinv.2025.104201

Mercilena Benjamin , Jay Singh , Avanish Kumar Pandey , Neha Thukral , Sarita Kumari , Jayanth Kumar Palanichamy , Sameer Bakhshi , Deepam Pushpam , Akash Kumar , Aditya Kumar Gupta , Jagdish Prasad Meena , Amitabh Singh , Pranay Tanwar , Amar Ranjan Singh , Sherry Bhalla , Anita Chopra

Whole-transcriptomic sequencing (WTS) has remarkably advanced our understanding of B-lineage acute lymphoblastic leukemia (B-ALL), allowing for detailed gene expression profiling and discovery of novel therapeutically relevant subtypes. The aim of this study was to evaluate the diagnostic and prognostic relevance of combining WTS with traditional genetic methods in risk-stratifying B-ALL. In a cohort of 394 patients (301 children and 93 adults), conventional techniques such as fluorescence in situ hybridization, cytogenetics, and reverse-transcription PCR identified sentinel chromosomal abnormalities like BCR::ABL1, TCF3::PBX1, ETV6::RUNX1, and KMT2A-R (rearranged), and ploidy status. WTS was performed on selected 257 patients to identify subtypes such as Ph-like, DUX4-R, PAX5-altered (PAX5-ALT), MEF2D-R, BCL2-R, UBTF-R, PAX5 P80R, NUTM1-R, ZNF384-R, ZNF384-like, ETV6::RUNX1-like, IKZF1 N159Y, and HLF-R. We used a multipronged strategy to identify the borderline subtypes such as Ph-like, PAX5-ALT, and CRLF2 (non–Ph-like), by integrating gene expression signatures using t-distributed stochastic neighbor embedding, subtype-defining mutations, gene fusions, and copy number assessments. Our integrated approach not only identifies prognostically relevant sentinel molecular subtypes but also increases subtype assignment in upto ∼95% of B-ALL patients. The pro-B immunophenotype was found to be more frequent in UBTF-R and MEF2D-R ALL. Ph-like ALL was associated with poor remission rates and higher minimal residual disease positivity, while DUX4-R showed favorable prognosis. We further categorized pediatric patients into 3 risk groups: favorable (hyperdiploid, ETV6::RUNX1, and DUX4-R), poor (BCR::ABL1, Ph-like, KMT2A-R, TCF3::PBX1, iAMP21, and hypodiploid), and intermediate (PAX5-ALT, PAX5 P80R, NUTM1-R, MEF2D-R, CRLF2 [non-Ph-like], UBTF-R, ZNF384-R, ZNF384-like, BCL2-R, IKZF1 N159Y, ETV6::RUNX1-like, and B-rest). Event-free survival and overall survival were significantly associated with this risk stratification. In adults, Ph-like ALL showed worse prognosis, particularly, in BCR::ABL1-negative ALL patients. Among the DUX4-R B-ALL, those with IKZF1 deletion had worse event-free survival and overall survival. We also identified several novel gene rearrangements in different subtypes of B-ALL. Our study demonstrated that integrating WTS with traditional methods provides a comprehensive, accurate, and cost-effective strategy for risk assessment and treatment planning for B-ALL.

全转录组测序（WTS）显著提高了我们对b系急性淋巴细胞白血病（B-ALL）的理解，允许详细的基因表达谱和发现新的治疗相关亚型。本研究的目的是评估WTS与传统遗传方法在B-ALL风险分层中的诊断和预后相关性。在394名患者（301名儿童和93名成人）的队列中，传统技术如FISH、细胞遗传学和RT-PCR鉴定出前哨染色体异常，如BCR::ABL1、TCF3::PBX1、ETV6::RUNX1和KMT2A-R（重排），以及倍性状态。选择257例患者进行WTS，鉴定Ph-like、DUX4-R、PAX5-altered （PAX5- alt）、MEF2D-R、BCL2-R、UBTF-R、PAX5 P80R、NUTM1-R、ZNF384-R、ZNF384-like、ETV6::RUNX1-like、IKZF1 N159Y和HLF-R等亚型。我们使用了多管齐下的策略，通过使用tSNE、亚型定义突变、基因融合和拷贝数评估整合基因表达特征，来识别边缘亚型，如Ph-like、PAX5-ALT和CRLF2（非Ph-like）。我们的综合方法不仅可以识别与预后相关的前哨分子亚型，还可以增加高达95%的B-ALL患者的亚型分配。前b免疫表型在UBTF-R和MEF2D-R ALL中更为常见。ph样ALL与较差的缓解率和较高的最小残留病阳性相关，而DUX4-R显示良好的预后。我们进一步将儿童患者分为三个风险组：良好（超二倍体，ETV6::RUNX1, DUX4-R），差（BCR::ABL1, Ph-like, KMT2A-R, TCF3::PBX1， iAMP21和次二倍体），和中间（PAX5- alt, PAX5 P80R, NUTM1-R, MEF2D-R, CRLF2（非Ph-like）， UBTF-R, ZNF384-R, ZNF384-like, BCL2-R, IKZF1 N159Y， ETV6::RUNX1-like和B-rest）。EFS和OS与这种风险分层显著相关。成人ph样ALL预后较差，尤其是BCR::ABL1阴性ALL患者。在DUX4-R B-ALL中，IKZF1缺失者的EFS和OS较差。我们还在不同的B-ALL亚型中发现了一些新的基因重排。我们的研究表明，将WTS与传统方法相结合，为B-ALL的风险评估和治疗计划提供了全面、准确、经济的策略。

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引用次数: 0

A Validation Workflow for Novel Oligonucleotide Sequences to Expand the Multiplexing Capacity of the CO-Detection by indEXing (CODEX) Platform 新寡核苷酸序列的验证工作流程，以扩大CODEX平台的多路复用能力。

IF 5.1 2区医学 Q1 MEDICINE, RESEARCH & EXPERIMENTAL

Laboratory Investigation

Pub Date : 2025-05-28 DOI: 10.1016/j.labinv.2025.104200

Alexander Rochwarger , Louisa Kaufmann , Jing Zhao , Ahmad Makky , Nhat Anh Nguyen , Nastassja Lehmann , Nikolay Samusik , Christine Beschorner , Saumya S. Manmadhan , Karen Greif , Christian M. Schürch

Antibody-based multiplexed tissue imaging has the potential to provide critical advances in biological discoveries and their translation for clinical applications. With the increasing introduction of markers and modalities for spatial analysis, there is an according demand for the expansion of multiplexing capacities of such imaging platforms. CO-Detection by indEXing (CODEX) is a widely used multiplexed tissue imaging platform that utilizes DNA-conjugated antibodies for imaging. The multiplexing capacity of CODEX is limited by the availability of unique DNA-oligonucleotide sequences for antibody barcoding. In this study, we demonstrate a workflow for the validation and the introduction of novel sets of candidate DNA-oligonucleotide sequences for CODEX. Through cross-validation multicycle experiments with the already published library of DNA barcodes, we here present a set of 27 novel oligonucleotide sequences for CODEX, increasing the potential multiplexing capacity to 85+ markers. We confirmed the utility of the new barcodes using a 74-plex antibody panel on a multitumor tissue microarray of paraffin-embedded tissues. The workflow presented here provides a reproducible method for extending the plexity of the CODEX platform, facilitating a deeper understanding of tissue microenvironments.

基于抗体的多路组织成像有可能为生物学发现及其临床应用的转化提供关键进展。随着越来越多地引入空间分析的标记和模式，对这种成像平台的多路复用能力的扩展有相应的需求。CODEX是一种广泛使用的多路组织成像平台，它利用dna偶联抗体进行成像。CODEX的多路复用能力受到用于抗体条形码的独特DNA寡核苷酸序列的可用性的限制。在这项研究中，我们展示了验证和引入新的候选DNA寡核苷酸序列的工作流程。通过与已发表的DNA条形码库的交叉验证多周期实验，我们在这里提出了一组27个新的CODEX寡核苷酸序列，将潜在的多路复用能力提高到85+个标记。我们通过在石蜡包埋组织的多肿瘤组织微阵列上使用74-plex抗体面板证实了新条形码的实用性。本文提出的工作流程为扩展CODEX平台的复杂性提供了一种可重复的方法，促进了对组织微环境的更深入了解。

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引用次数: 0

Expression of Homo Sapiens (Hsa)-miR-139-5p as a Clinically Feasible Prognostic Marker for Differentiated Thyroid Cancer 智人(Hsa)-miR-139-5p的表达作为分化型甲状腺癌（DTC）的临床可行预后标志物

IF 5.1 2区医学 Q1 MEDICINE, RESEARCH & EXPERIMENTAL

Laboratory Investigation

Pub Date : 2025-05-27 DOI: 10.1016/j.labinv.2025.104199

Natalia Martínez-Puente , Ignacio Ruz-Caracuel , Luis J. Leandro-García , Héctor Pian-Arias , Zaira Vega-Corral , Rocío Letón , Roberta Radu , Mariolga Berrizbeitia , Sara Mellid , Clara Reglero , Milton E. Salazar-Hidalgo , Ester Arroba , Alberto Díaz-Talavera , Mónica Marazuela , Amparo Benito-Berlinches , Irene González-García , Sandra Campos-Mena , María D. Lozano-Escario , Sonsoles Guadalix , María Calatayud , Cristina Montero-Conde

Prognostic uncertainty often leads to overtreatment of differentiated thyroid cancer (DTC) or unspecific management of more aggressive entities. MiR-139-5p (miR-139-5p) has emerged as a promising prognostic factor that may enhance current individual risk assessment systems. Therefore, we aimed to validate miR139-5p expression as a prognostic marker in DTC using a standardized method and to establish expression cutoff values for discriminating prognostic groups. In addition, we explored an in situ approach to analyze this microRNA expression as a potential molecular tool for clinical practice. We collected a tissue series of 132 samples, including thyroid tumors, adjacent normal thyroid tissue, and lymph node metastases from a long-term follow-up retrospective cohort of 60 patients with DTC with either progressive/persistent disease or an excellent response to primary treatment. We first identified recurrent tumor driver mutations and TERT promoter mutations using next-generation sequencing. Through a standardized paired tumor/normal qPCR analysis, we confirmed a significant reduction in miR139-5p expression in progressive/persistent DTCs compared with excellent response DTCs (P value = .002). Further analysis, including thyroid cancer The Cancer Genome Atlas tumor/normal pairs (n = 59), showed a strong association between reduced miR139-5p expression and TERT promoter mutations (P < .001), as well as advanced local or distant metastasis at diagnosis (P = .031). Next, we established miR139-5p^HIGH and miR139-5p^LOW tumor/normal cutoff values to discriminate prognostic groups, with high expression predicting excellent response and low expression predicting disease progression/persistence. Cutoff values were defined through logistic regression and receiver operating characteristic curve analysis and validated in an independent cohort (n = 38). Quantitative image analysis using QuPath software of an automatic chromogenic in situ hybridization assay for miR139 detection further supported the qPCR findings and revealed heterogeneous intratumor miR139 expression, which was lowest in the Ki-67 proliferation index--positive foci. Overall, our data indicate that miR139 expression assessment is a feasible tool for clinical use, potentially reducing overtreatment during primary DTC interventions and supporting a risk-adjusted follow-up schedule.

预后的不确定性常常导致分化型甲状腺癌（DTC）的过度治疗或对更具侵袭性实体的不特异性治疗。MiR-139-5p （miR139-5p）已成为一个有希望的预后因素，可以增强当前的个人风险评估系统。因此，我们旨在使用标准化方法验证miR139-5p表达作为DTC的预后标志物，并建立用于区分预后组的表达临界值。此外，我们探索了一种原位方法来分析这种miRNA表达，作为临床实践的潜在分子工具。我们收集了132个组织样本，包括甲状腺肿瘤、邻近正常甲状腺组织和淋巴结转移，这些样本来自60名进展性/持续性疾病或对初级治疗有良好反应的DTC患者的长期随访回顾性队列。我们首先使用下一代测序技术确定了复发性肿瘤驱动突变和TERT启动子突变。通过标准化的配对肿瘤/正常qPCR分析，我们证实了进展性/持续性dtc中miR139-5p的表达与优异反应性dtc相比显著降低（p值= 0.002）。进一步的分析，包括甲状腺癌TCGA肿瘤/正常对（n = 59），显示miR139-5p表达降低与TERT启动子突变（p值< 0.001）以及诊断时的晚期局部或远处转移（p值= 0.031）之间有很强的相关性。接下来，我们建立了miR139-5pHIGH和miR139-5pLOW肿瘤/正常切断值来区分预后组，高表达预测良好的反应，低表达预测疾病进展/持续。通过逻辑回归和ROC曲线分析确定截断值，并在独立队列（n = 38）中进行验证。使用QuPath软件对自动显色原位杂交检测miR139的定量图像分析进一步支持了qPCR的发现，并显示肿瘤内miR139的表达具有异质性，在Ki-67增殖指数阳性灶中表达最低。总的来说，我们的数据表明，miR139表达评估是一种可行的临床应用工具，有可能减少原发性DTC干预期间的过度治疗，并支持风险调整的随访计划。

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引用次数: 0

Decoding the Molecular Landscape of 262 Uterine Sarcomas: RNA-Seq Clustering of Endometrial Stromal Sarcomas, Uterine Tumors Resembling Ovarian Sex Cord Tumors, and Undifferentiated Uterine Sarcomas With Prognostic Insights 解码262个子宫肉瘤的分子景观：ESS， UTROSCT和UUS的RNA-Seq聚类与预后见解。

IF 5.1 2区医学 Q1 MEDICINE, RESEARCH & EXPERIMENTAL

Laboratory Investigation

Pub Date : 2025-05-24 DOI: 10.1016/j.labinv.2025.104198

Jan Hojný , Jiří Dvořák , Romana Vránková , Michaela Kendall Bártů , Nikola Hájková , Eva Krkavcová , Miroslava Flídrová , Ivana Stružinská , Kristýna Němejcová , Jiří Bouda , Květoslava Michalová , David Cibula , Renata Poncová , Janusz Rys , Mariusz Ksiazek , Marcin Jędryka , Tymoteusz Poprawski , Alberto Berjón , Ignacio Zapardiel , Jan Laco , Pavel Dundr

Low-grade endometrial stromal sarcomas (LG-ESS), high-grade ESS (HG-ESS), undifferentiated uterine sarcomas (UUS), and uterine tumors resembling ovarian sex cord tumors are distinct non-smooth muscle cell neoplasms with varying clinical outcomes, often exhibiting overlapping characteristics. Diagnosis can be supported by identifying characteristic recurrent translocations, which may be absent in some cases, complicating the distinction of equivocal cases. Additionally, cases with overlapping features of low-grade and high-grade characteristics are recognized. To address these challenges, we analyzed RNA-seq profiles of 262 cases. Our results revealed that LG-ESS, with and without recurrent fusions, clustered into 2 partially overlapping expression profiles associated with distinct overall and relapse-free survival outcomes, with the cluster containing a majority of fusion-negative tumors demonstrating better prognoses. uterine tumors resembling ovarian sex cord tumors expression profiles closely resembled those of both LG-ESS subgroups, with NCOA3 fusion-positive cases clustering in groups with better survival outcomes. Furthermore, a distinct cluster for HG-ESS with BCOR and YWHAE fusions was identified, differentiating these tumors from HG-ESS without fusions. ONECUT3 emerged as a potential specific marker for this HG-ESS-fusion entity. A significant expression overlap was observed between monomorphic HG-ESS without fusions and pleomorphic UUS. These samples separated further into 2 mixed clusters distinguished by differences in immune activity, which significantly influenced overall survival and relapse-free survival outcomes. Unsupervised clustering of UUS revealed subgroups resembling either HG-ESS or muscle-cell–differentiated tumors, suggesting that UUS may include poorly differentiated distinct entities, such as leiomyosarcoma, and that the distinction from HG-ESS may, in some cases, be arbitrary. Our transcriptome analysis highlights several entities with distinct survival characteristics, providing a foundation for further characterization of these rare, often difficult-to-classify, tumors.

低级别子宫内膜间质肉瘤（LG-ESS）、高级别子宫内膜间质肉瘤（HG-ESS）、未分化子宫肉瘤（UUS）和类似卵巢性索肿瘤的子宫肿瘤（UTROSCT）是不同的非平滑肌细胞肿瘤，临床结果不同，通常表现出重叠特征。诊断可以通过识别特征性的复发性易位来支持，在某些情况下可能不存在易位，使模棱两可病例的区分复杂化。此外，还可以识别出LG和HG特征重叠的病例。为了解决这些挑战，我们分析了262例病例的RNA-Seq谱。我们的研究结果显示，有或没有复发性融合的LG-ESS聚集成两个部分重叠的表达谱，与不同的总体和无复发生存结果相关，其中包含大多数融合阴性肿瘤的集群表现出更好的预后。UTROSCT表达谱与两个LG-ESS亚组非常相似，NCOA3融合阳性病例聚集在生存结果较好的组中。此外，鉴定出具有bor和YWHAE融合的HG-ESS的独特集群，将这些肿瘤与没有融合的HG-ESS区分开来。ONECUT3作为这种hg - ess融合实体的潜在特异性标记物出现。在未融合的单态HG-ESS和多形性UUS之间观察到显著的表达重叠。这些样本进一步分成两个混合簇，以免疫活性差异区分，这显著影响总体生存和无复发生存结果。UUS的无监督聚类揭示了类似于HG-ESS或肌肉细胞分化肿瘤的亚群，表明UUS可能包括低分化的不同实体，如平滑肌肉瘤，并且在某些情况下与HG-ESS的区分可能是任意的。我们的转录组分析突出了几个具有不同生存特征的实体，为进一步表征这些罕见的、通常难以分类的肿瘤提供了基础。

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引用次数: 0

Aryl Hydrocarbon Receptor Deficiency Upregulates Intercellular Adhesion Molecule 1 in Retinal Pigment Epithelial Cells and Contributes to Retinal Inflammation 芳香烃受体缺乏可上调视网膜色素上皮细胞的细胞间粘附分子1，导致视网膜炎症。

IF 5.1 2区医学 Q1 MEDICINE, RESEARCH & EXPERIMENTAL

Laboratory Investigation

Pub Date : 2025-05-21 DOI: 10.1016/j.labinv.2025.104197

Tsung-Min Yang , Te-Chao Fang , Yu-Cheng Lee , Chen-Chen Lee , Yen-Ju Chan , Ida Fitriana , Yu-Wen Cheng , Ching-Hao Li

Retinal pigment epithelium (RPE) cells, located between the photoreceptors and choroid, play a crucial role in maintaining retinal health and function. They act as immunosuppressive barriers, preventing immune cell infiltration from the choroid. Retinal inflammation contributes to the development of various ocular diseases. The aryl hydrocarbon receptor (AHR) is a well-established ligand-dependent transcription factor that mediates potent anti-inflammatory signals following ligand binding. AHR expression is notably reduced under several conditions that negatively affect the retina. We hypothesized that AHR protein loss may impair RPE cell function, shifting them toward a proinflammatory phenotype. In this study, we investigated the proinflammatory pathways activated by AHR knockout (AHR-KO) and examined associated retinal phenotypic changes in AHR-KO mice. Our findings suggest that AHR deficiency may enhance the activity of αvβ3-integrin, extracellular signal–regulated kinase 1/2, and p65 subunit of nuclear factor kappa B, leading to an upregulation of intercellular adhesion molecule 1 (ICAM1) and promoting monocyte adhesion in vitro. Introducing an AHR-green fluorescent protein into AHR-KO RPE cells or pretreating the cells with pharmacologic inhibitors targeting αvβ3 (cyclo[RGDfk]), focal adhesion kinase (PF573228), phospholipase C (U73122), extracellular signal–regulated kinase 1/2 (U0126), and nuclear factor kappa B (Bay11-7082) prevented ICAM1 induction in AHR-KO RPE cells. These results suggest that the proinflammatory pathway is driven by AHR deficiency. In AHR-KO mice, retinal tissues showed ICAM1 accumulation, microglial activation, and migration, indicating chronic retinal inflammation because of AHR deficiency. These mice also displayed early-onset electroretinogram degeneration. Collectively, our data support the protective role of AHR in maintaining RPE cell physiology and retinal health.

视网膜色素上皮细胞（RPE）位于光感受器和脉络膜之间，在维持视网膜健康和功能中起着至关重要的作用。它们作为免疫抑制屏障，阻止免疫细胞从脉络膜渗透。视网膜炎症有助于各种眼部疾病的发展。芳烃受体（AHR）是一种成熟的配体依赖性转录因子，在配体结合后介导有效的抗炎信号。在几种对视网膜有负面影响的情况下，AHR的表达明显减少。我们假设AHR蛋白丢失可能损害RPE细胞功能，使其向促炎表型转变。在这项研究中，我们研究了AHR敲除（AHR- ko）激活的促炎途径，并检查了AHR- ko小鼠的相关视网膜表型变化。我们的研究结果表明，AHR缺乏可能会增强αvβ3整合素、细胞外信号调节激酶（ERK1/2）和核因子κB p65亚基（NF-κB）的活性，导致细胞间粘附分子1 （ICAM1）的上调，促进单核细胞的体外粘附。将ahr -绿色荧光蛋白导入AHR-KO RPE细胞或用靶向αvβ3 （cycloRGDfk）、局灶黏附激酶（PF573228）、磷脂酶C （U73122）、ERK1/2 （U0126）和NF-κB （Bay11-7082）的药物抑制剂预处理细胞可阻止ICAM1在AHR-KO RPE细胞中的诱导。这些结果表明促炎途径是由AHR缺乏驱动的。在AHR- ko小鼠中，视网膜组织显示ICAM1积累、小胶质细胞激活和迁移，表明AHR缺乏导致慢性视网膜炎症。这些小鼠也表现出早发性视网膜电图变性。总的来说，我们的数据支持AHR在维持RPE细胞生理和视网膜健康方面的保护作用。

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引用次数: 0