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 患者衍生的肿瘤器官（PDO），涉及新鲜肿瘤片段的体内培养物， 已经成为预测个性化癌症患者药物反应的有前途的模型 治疗。 PDO概括肿瘤微环境（TME），类似于源肿瘤 表型和基因组上，与高通量药物筛查兼容。 但是，缺乏PDO中的免疫细胞保存一直是 建模免疫疗法。我们的团队最近展示了一种培养肿瘤的新型PDO 片段作为凝聚力单元，允许原位保存各种免疫细胞类型 与没有人造重构的肿瘤细胞一起。这种方法使 患者对免疫检查点抑制剂的特定反应。 PDOS产生的第一步之一是解剖患者肿瘤标本 变成小碎片。机械解剖，而不是酶消化，至关重要 保留肿瘤细胞与内源性免疫和非免疫的体内关联 元素。保留内源性免疫细胞的能力，包括肿瘤浸润 淋巴细胞（TIL）对于个性化免疫疗法测试尤为重要。 但是，当前的机械解剖主要依赖于肿瘤标本的手动切割 变成小碎片。它导致尺寸范围较大的碎片，并且是不精确的，并且 不可重复的。太大的营养供应不足的碎片，次优的碎片 氧合和生存能力以及药物渗透不良。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 可能以低浓度存在。 因此，有一种未满足的需要一种更好的方法来产生可控的肿瘤片段 和均匀的尺寸，并确定最佳尺寸以提高可行的可重复性和产量 可以保留细胞上下文和肿瘤结构的PDO。这个项目旨在 通过开发一种新方法将肿瘤标本解剖为 均匀的碎片。性能指标包括碎片尺寸均匀性，PDO生存能力， 免疫细胞的保存和肿瘤细胞毒性响应免疫疗法。其他切割 包括手动切割在内的方法将用作基准。