Studying Immunogenic Cell Death in Human Colorectal Cancer Organoids.

IF 2.7 4区医学 Q3 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

OncoTargets and therapy Pub Date : 2025-06-06 eCollection Date: 2025-01-01 DOI:10.2147/OTT.S521346

Rebecca Lisandrelli, Matthias Winkler, Zlatko Trajanoski, Giorgia Lamberti

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引用次数: 0

Abstract

Purpose: Combination therapies of chemotherapeutic agents and immunotherapy have shown promising results in the treatment of cold tumors. Various chemotherapies trigger immunogenic cell death (ICD) and release of hallmarks immunogenic damage associated molecular patterns (DAMPs) that have been related with immunostimulatory activities, leading to better patient prognosis. We aim to optimize in vitro assays to detect DAMPs release in response to chemotherapeutic agents using a colorectal cancer (CRC) organoids model.

Methods: CRC patient-derived organoids (PDOs) were treated either with oxaliplatin (OXA) or with 5-fluorouracil (5FU) and viability was measured with CellTiter-Glo3D Cell viability assay. Calreticulin (CALR) and high mobility group box 1 protein (HMGB1) intracellular translocation was assessed in immunofluorescence microscopy and quantified via co-localization with wheat germ agglutinin (WGA) and DAPI. Extracellular release of adenosine triphosphate (ATP) was quantified via specific luminescence assays.

Results: The results showed that CRC PDOs release DAMPs in a patient-specific manner in response to OXA and 5FU treatments.

Conclusion: This study successfully used immunofluorescence and luminescence methods to detect ICD-associated DAMPs release in CRC PDOs in response to chemotherapeutic treatments. This approach allows the recognition of patient-specific ICD activation and could help predict patient response to therapies.

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人类结直肠癌类器官免疫原性细胞死亡的研究

目的：化疗药物与免疫疗法联合治疗感冒肿瘤已显示出良好的效果。各种化疗触发免疫原性细胞死亡（ICD）和释放与免疫刺激活动相关的标志性免疫原性损伤相关分子模式（DAMPs），导致更好的患者预后。我们的目标是利用结直肠癌（CRC）类器官模型优化体外检测DAMPs释放对化疗药物反应的方法。方法：用奥沙利铂（OXA）或5-氟尿嘧啶（5FU）治疗结直肠癌患者源性类器官（PDOs），用CellTiter-Glo3D细胞活力测定法测定其活力。采用免疫荧光显微镜检测钙网蛋白（CALR）和高迁移率组盒蛋白（HMGB1）的细胞内易位，并与小麦胚芽凝集素（WGA）和DAPI共定位。三磷酸腺苷（ATP）的细胞外释放通过特异性发光测定。结果：结果显示CRC PDOs以患者特异性方式释放DAMPs，以响应OXA和5FU治疗。结论：本研究成功地利用免疫荧光和发光方法检测了icd相关DAMPs在结直肠癌PDOs中对化疗的释放。这种方法可以识别患者特异性ICD激活，并有助于预测患者对治疗的反应。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

OncoTargets and therapy BIOTECHNOLOGY & APPLIED MICROBIOLOGY-ONCOLOGY

CiteScore

9.70

自引率

0.00%

发文量

221

审稿时长

1 months

期刊介绍： OncoTargets and Therapy is an international, peer-reviewed journal focusing on molecular aspects of cancer research, that is, the molecular diagnosis of and targeted molecular or precision therapy for all types of cancer. The journal is characterized by the rapid reporting of high-quality original research, basic science, reviews and evaluations, expert opinion and commentary that shed novel insight on a cancer or cancer subtype. Specific topics covered by the journal include: -Novel therapeutic targets and innovative agents -Novel therapeutic regimens for improved benefit and/or decreased side effects -Early stage clinical trials Further considerations when submitting to OncoTargets and Therapy: -Studies containing in vivo animal model data will be considered favorably. -Tissue microarray analyses will not be considered except in cases where they are supported by comprehensive biological studies involving multiple cell lines. -Biomarker association studies will be considered only when validated by comprehensive in vitro data and analysis of human tissue samples. -Studies utilizing publicly available data (e.g. GWAS/TCGA/GEO etc.) should add to the body of knowledge about a specific disease or relevant phenotype and must be validated using the authors’ own data through replication in an independent sample set and functional follow-up. -Bioinformatics studies must be validated using the authors’ own data through replication in an independent sample set and functional follow-up. -Single nucleotide polymorphism (SNP) studies will not be considered.