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: 10705736
• 负责人: Janis M Taube
• 金额: $ 19.19万
• 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/10705736
• 关键词: Advanced Development; Age; Algorithmic Analysis; Algorithms; Antibodies; Antigens; Applications Grants; Biological Assay; Biological Markers; Biological Sciences; Biopsy; CD8B1 gene; Cancer Center; Cd68; Cell surface; Cells; 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; biomarker development; biomarker identification; cell type; clinic ready; 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; spectrograph; 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.

项目概要 预处理活检中 PD-L1 膜（细胞表面）表达最多 常用的抗 PD-1 治疗反应可能性的相关性。一般发现 PD-L1 表达与肿瘤对抗 PD-1 治疗的反应之间存在关联 在数万名接受抗肿瘤药物治疗的患有多种肿瘤类型的患者中得到证实 PD-(L)1。然而，虽然 PD-L1 表达丰富以对抗 PD-(L)1 的反应，但并非如此。 充足的。此外，虽然病理学家证明肿瘤细胞评分具有良好的重现性 (TC) 通过显色 IHC 和光学显微镜检测 PD-L1 表达，重现性较差 用于对免疫细胞 (IC) 上的 PD-L1 表达进行评分。 其他已被证明可以改善 PD-L1 生物标志物的相关功能包括 PD-1 与 PD-L1 的接近度、CD8+FoxP3+ 细胞的密度以及 CD68 和肿瘤标志物 免疫染色，后者有助于识别 IC 和 TC 上 PD-L1 的共表达， 分别。捕获所有这些的定量多重免疫荧光测定 已开发出功能，包括 PD-L1、PD-1、CD8、FoxP3、CD68（肿瘤标志物） （细胞角蛋白 AE1/3 用于非小细胞肺癌，NSCLC），一种泛膜标记物和 DAPI。通过这种测定，可以枚举关键的细胞亚群及其共表达 配置文件。还可以包括空间参数，包括 PD-1 和 PD-L1，以前未曾被纳入预测或预后手术病理学中 基于样本的分析。该 mIF 测定提高了响应的灵敏度和特异性 与多肿瘤中单独的 PD-L1 表达评估相比，抗 PD1 疗法 类型，包括黑色素瘤、非小细胞肺癌和默克尔细胞癌等。 本提案的目的是对 mIF 染色测定进行位点间验证 以及四个主要学术站点（约翰霍普金斯大学，医学博士）之间的相关锁定算法 安德森、耶鲁大学和普罗维登斯波特兰医学中心）。以下分析 将使用来自 250 名晚期 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），与以下临床结果相关 基于抗 PD-1 的治疗。该研究的成果是一种精细的、多重生物标志物测定 对抗 PD-1 的反应/耐药性已在多个学术场所得到验证。这 结果将是一种多重 IF 测定，适合进行高级开发，旨在 临床实施。虽然非小细胞肺癌是当前拨款提案的重点，但初步 结果表明，该测定对于许多其他实体瘤类型也具有巨大价值。

项目成果

Comparing and Correcting Spectral Sensitivities between Multispectral Microscopes: A Prerequisite to Clinical Implementation.

比较和校正多光谱显微镜之间的光谱灵敏度：临床实施的先决条件。

• 发表时间: 2023-06-08
• 影响因子: 5.2
• 作者: Eminizer, Margaret;Nagy, Melinda;Engle, Elizabeth L;Soto;Jorquera, Andrew;Roskes, Jeffrey S;Green, Benjamin F;Wilton, Richard;Taube, Janis M;Szalay, Alexander S
• 通讯作者: Szalay, Alexander S