Analytic and Clinical Validation of a 7-plex MIF Assay for Predictive Response of Advanced NSCLC to Anti-PD-1 based Therapy

7 重 MIF 检测对晚期 NSCLC 对抗 PD-1 治疗的预测反应的分析和临床验证

• 批准号: 10506186
• 负责人: Janis M Taube
• 金额: $ 22.43万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-16 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10506186
• 关键词: Advanced Development; Age; Algorithmic Analysis; Algorithms; Antibodies; Antigens; Applications Grants; Archives; Biological Assay; Biological Markers; Biological Sciences; Biopsy; CD8B1 gene; Cancer Center; Cell surface; Cells; Clinic; Clinical; Collaborations; Cytokeratin; Disease; Evaluation; FOXP3 gene; Formalin; Gender; Geography; Goals; Image Analysis; Immune; Immunofluorescence Immunologic; Immunohistochemistry; Immunooncology; Individual; Institution; Label; Link; Malignant - descriptor; Malignant Epithelial Cell; Medical center; Membrane; Merkel cell carcinoma; Modality; Morphologic artifacts; Non-Small-Cell Lung Carcinoma; Outcome; PD-1 pathway; PD-1/PD-L1; PD-L1 blockade; Paraffin Embedding; Participant; Pathologist; Patient-Focused Outcomes; Patients; Pigments; Positioning Attribute; Reagent; Receiver Operating Characteristics; Reproducibility; Research Personnel; Resistance; Resolution; Sensitivity and Specificity; Site; Slide; Solid Neoplasm; Specific qualifier value; Specimen; Stains; Standardization; Surgical Pathology; Testing; Therapeutic Agents; Tissue Embedding; Tissues; Tonsil; Tumor Markers; Tumor stage; Universities; Update; Validation; Work; age related; anti-PD-1; anti-PD-L1; anti-PD1 therapy; base; biomarker development; cell type; clinical care; clinical implementation; cohort; combinatorial; companion diagnostics; density; design; imaging approach; imaging platform; immunotherapy clinical trials; improved; indexing; light microscopy; liquid crystal polymer; melanoma; multiparametric imaging; multiplex assay; neoplastic cell; next generation; phenotypic data; predicting response; predictive test; prognostic; programmed cell death ligand 1; programmed cell death protein 1; research clinical testing; response; tumor; tumor microenvironment; whole slide imaging

PROJECT SUMMARY PD-L1 membranous (cell surface) expression in pretreatment biopsies is the most commonly used correlate of the likelihood of response to anti-PD-1 therapy. The general finding of an association of PD-L1 expression with tumor response to anti-PD-1 therapy has been substantiated across tens of thousands of patients with numerous tumor types treated with anti- PD-(L)1. However, while PD-L1 expression enriches for response to anti-PD-(L)1, it is not sufficient. Additionally, while pathologists demonstrate good reproducibility for scoring tumor cell (TC) PD-L1 expression by chromogenic IHC and light microscopy, they have poor reproducibility for scoring PD-L1 expression on immune cells (IC). Other related features which have been shown to improve on the PD-L1 biomarker include the proximity of PD-1 to PD-L1, the density of CD8+FoxP3+ cells, and CD68 and tumor marker immunostains, the latter of which help identify co-expression of PD-L1 on ICs and TCs, respectively. A quantitative multiplex immunofluorescence assay which captures all of these features has been developed and includes PD-L1, PD-1, CD8, FoxP3, CD68, a tumor marker (cytokeratin AE1/3 for non-small cell lung carcinoma, NSCLC), a pan-membrane marker and DAPI. Through this assay, it is possible to enumerate key cellular subsets and their co-expression profiles. It is also possible to include spatial parameters, including the distance between PD-1 and PD-L1, which have not previously been included in predictive or prognostic surgical pathology specimen-based assays. This mIF assay has increased sensitivity and specificity for response to anti-PD1 therapy when compared to the assessment of PD-L1 expression alone in multiple tumor types, including melanoma, NSCLC, and Merkel cell carcinoma, amongst others. The purpose of this proposal is to perform inter-site validation of the mIF staining assay and associated lock-down algorithm amongst four major academic sites (Johns Hopkins, MD Anderson, Yale University, and Providence Portland Medical Center). Following analytical validation, discovery and validation cohorts from 250 patients with advanced NSCLC will be used to establish final assay parameters (including thresholds), linked to clinical outcomes following anti-PD-1-based therapy. The deliverable of the study is a refined, multiplex biomarker assay for response/resistance to anti-PD-1 that has been validated across multiple academic sites. The result will be a multiplex IF assay that is suitably staged for advanced development aimed at clinical implementation. While NSCLC is the focus of the current grant proposal, preliminary results suggest that this assay will also have great value in numerous other solid tumor types.

项目摘要 预处理活检中的PD-L1膜（细胞表面）表达最多 通常使用抗PD-1治疗的可能性的相关性。一般发现 PD-L1表达与肿瘤对抗PD-1治疗的反应的关联已是 在数以万计的患有抗肿瘤类型的患者中得到证实 pd-（l）1。但是，虽然PD-L1表达富集以响应抗PD-（L）1，但不是 充足的。另外，尽管病理学家证明了评分肿瘤细胞的良好可重复性 （TC）PD-L1通过成色IHC和光学显微镜表达，它们的可重复性差 用于在免疫细胞上评分PD-L1表达（IC）。 已显示在PD-L1生物标志物上改进的其他相关功能包括 PD-1与PD-L1的接近度，CD8+ FOXP3+细胞的密度以及CD68和肿瘤标记 免疫剂，后者有助于确定PD-L1在ICS和TCS上的共表达， 分别。定量多重免疫荧光测定法，捕获所有这些 已经开发了功能，包括PD-L1，PD-1，CD8，FOXP3，CD68，肿瘤标记 （非小细胞肺癌，NSCLC的细胞角蛋白AE1/3），泛包膜标记和 DAPI。通过此测定，可以枚举钥匙细胞集及其共表达 概况。还可以包括空间参数，包括PD-1和 PD-L1，以前尚未包含在预测性或预后手术病理中 基于标本的测定。该MIF分析具有提高的灵敏度和对响应的特异性 与单独评估PD-L1表达在多个肿瘤中的评估相比，抗PD1治疗 包括黑色素瘤，NSCLC和默克尔细胞癌等类型。 该提案的目的是对MIF染色测定法执行现场验证 以及相关的四个主要学术网站中相关的锁定算法（约翰·霍普金斯（Johns Hopkins，MD） 安德森，耶鲁大学和普罗维登斯波特兰医学中心）。分析后 将使用250名高级NSCLC患者的验证，发现和验证队列 建立最终测定参数（包括阈值），与临床结果相关联 基于抗PD-1的治疗。该研究的可交付是一种精致的多重生物标志物测定法 对抗PD-1的响应/抗性已在多个学术地点进行了验证。这 如果适当地进行高级开发的测定，则结果将是一个多路复用 临床实施。虽然NSCLC是当前赠款提案的重点，但初步 结果表明，该测定法在许多其他实体瘤类型中也将具有很高的价值。