A micro-dissection platform for generating uniform-sized patient-derived tumor organoids (PDOs) for personalized cancer therapy

一个显微解剖平台，用于生成大小一致的患者来源的肿瘤类器官（PDO），用于个性化癌症治疗

• 批准号: 10697348
• 负责人: Sindy Kam-Yan Tang
• 金额: $ 17.93万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-05 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10697348
• 关键词: Address; Adoption; Antineoplastic Agents; Architecture; B-Lymphocytes; Benchmarking; Biological; Cancer Biology; Cause of Death; Cells; Clinical; Cryopreservation; Data; Devices; Diffusion; Digestion; Dissection; Drug Screening; Elements; Fibroblasts; Flow Cytometry; Generations; Genomics; Geometry; Goals; Growth; Image; Immune; Immune checkpoint inhibitor; Immunofluorescence Immunologic; Immunotherapy; In Situ; Ischemia; Kidney; Lead; Lung; Macrophage; Malignant Neoplasms; Manuals; Measures; Mechanics; Methods; Microdissection; Microfabrication; Modeling; Natural Killer Cells; Nutrient; Organoids; Outcome; Oxygen; Patients; Penetration; Performance; Pharmaceutical Preparations; Phenotype; Play; Process; Quantitative Reverse Transcriptase PCR; Reproducibility; Role; Sampling; Silicon; Source; Specimen; Standardization; Stromal Cells; Surgical Pathology; T-Lymphocyte; Technology; Testing; Time; Tissue Banks; Tissues; Tumor Tissue; Tumor-Infiltrating Lymphocytes; Validation; Variant; Work; biobank; cancer genome; cell type; checkpoint inhibition; cytotoxicity; experimental study; high-throughput drug screening; in vivo; instrumentation; nanofabrication; neoplastic cell; novel; patient response; personalized cancer therapy; personalized immunotherapy; predictive modeling; preservation; reconstitution; response; tumor; tumor microenvironment; tumor progression

Project Summary Patient-derived tumor organoids (PDO), involving the ex vivo culture of fresh tumor fragments, have emerged as promising models for predicting patient drug response for personalized cancer therapy. PDOs recapitulate the tumor micro-environment (TME), resemble the source tumor phenotypically and genomically, and are compatible with high-throughput drug screening. However, the lack of preservation of immune cells in PDOs has been a major roadblock to modeling immunotherapy. Our team recently demonstrated a new type of PDO that cultures tumor fragments as a cohesive unit, allowing the in situ preservation of diverse immune cell types alongside tumor cells without artificial reconstitution. This approach has enabled the modeling of patient-specific responses to immune checkpoint inhibitors. One of the first steps in the generation of PDOs is the dissection of patient tumor specimen into small fragments. Mechanical dissection, instead of enzymatic digestion, is critical in preserving the in vivo association between tumor cells and endogenous immune and non-immune elements. The ability to preserve endogenous immune cells, including tumor-infiltrating lymphocytes (TIL), is particularly important for personalized immunotherapy testing. However, current mechanical dissection relies primarily on manual mincing of tumor specimen into small fragments. It results in fragments with a broad size range, and is imprecise and irreproducible. Fragments that are too large suffer from inadequate nutrient supply, suboptimal oxygenation and viability, and poor drug penetration. Fragments that are too small are unlikely to preserve sufficient stromal cells to support PDO growth, and/or endogenous immune cells which may be present at low concentrations. As such, there is an unmet need for a better way to generate tumor fragments of controllable and uniform size, and identify optimal size(s) to increase the reproducibility and yield of viable PDOs that can preserve the cellular contexture and tumor architecture. This project aims to address this need by developing a new method to mechanically dissect tumor specimen into uniform fragments. Performance measures include fragment size uniformity, PDO viability, preservation of immune cells, and tumor cytotoxicity in response to immunotherapy. Other cutting methods including manual mincing will be used as benchmarks.

项目摘要 患者来源的肿瘤有机化合物(PDO)，涉及新鲜肿瘤碎片的体外培养， 已经成为预测个性化癌症患者药物反应的有希望的模型 心理治疗。PDOS概括了肿瘤微环境(TME)，类似于来源肿瘤 表型和基因组，并与高通量药物筛选兼容。 然而，缺乏对PDO中免疫细胞的保存一直是阻碍 模范免疫疗法。我们的团队最近展示了一种新型的PDO，它可以培养肿瘤 片段作为一个凝聚单元，允许原位保存不同类型的免疫细胞 在没有人工重建的情况下与肿瘤细胞一起。这种方法使建模成为可能 患者对免疫检查点抑制剂的特定反应。 生成PDO的第一步是对患者肿瘤标本的解剖 变成了小碎片。机械解剖，而不是酶消化，是关键 保留体内肿瘤细胞与内源性免疫和非免疫的联系 元素。保护内源性免疫细胞的能力，包括肿瘤浸润性 淋巴细胞(TIL)对于个性化免疫治疗测试尤为重要。 然而，目前的机械解剖主要依赖于人工切碎肿瘤标本。 变成了小碎片。它导致碎片的大小范围很宽，并且不精确 是不可复制的。太大的碎片会受到营养供应不足的影响，不是最理想的 氧合能力和生存能力，以及药物渗透性差。太小的碎片不太可能 保存足够的基质细胞以支持PDO生长，和/或内源性免疫细胞 可能在低浓度下存在。 因此，存在对产生可控肿瘤片段的更好方法的未得到满足的需求 和均匀的大小，并确定最佳大小(S)，以增加重复性和产量的可行 可以保存细胞结构和肿瘤结构的PDO。该项目旨在 通过开发一种新的方法来机械地将肿瘤标本解剖成 制服碎片。性能指标包括碎片大小一致性、PDO生存能力、 免疫细胞的保存，以及免疫治疗反应中的肿瘤细胞毒性。其他切割 包括手动切碎在内的方法将被用作基准。