Evaluation of Patient Factors and Sample Pre-Analytics on Predictive Multiplex Immunohistochemical Assays in Immuno-Oncology Patients

免疫肿瘤患者预测多重免疫组织化学分析的患者因素和样品预分析的评估

• 批准号: 10907058
• 负责人: Michael H. A. Roehrl
• 金额: $ 20.11万
• 依托单位: BETH ISRAEL DEACONESS MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-01 至 2027-11-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10907058
• 关键词: 4 year old; Acceleration; Address; Affect; Age; Alcohols; Antiemetics; Asian; Biological Assay; Biological Markers; Biopsy; Black race; Body mass index; Cancer Patient; Categories; Characteristics; Clinic; Clinical; Collaborations; Cytotoxic Chemotherapy; Data; Databases; Dependence; Eastern Cooperative Oncology Group; Ensure; Ethnic Origin; Evaluation; Excision; Face; Formalin; Freezing; Gender; Goals; Guidelines; Health Care Costs; Hispanic; Ice; Immunohistochemistry; Immunooncology; Immunotherapy; Ischemia; Knowledge; Length; Lymphocyte; Malignant Neoplasms; Malignant neoplasm of lung; Malignant neoplasm of urinary bladder; Measurement; Measures; Memorial Sloan-Kettering Cancer Center; Molecular; Non-Small-Cell Lung Carcinoma; Outcome; Paraffin Tissue; Patients; Performance; Performance Status; Pharmacotherapy; Physiological; Procedures; Process; Prospective Studies; Prospective cohort; Proteins; Protocols documentation; Qualifying; Radiation; Research; Sampling; Site; Solid; Specificity; Specimen; Steroids; Temperature; Testing; Time; Tissue Sample; Tissues; Translations; Tumor Tissue; Variant; anti-PD-L1; biobank; cancer type; cell type; clinical practice; cohort; contextual factors; diagnostic assay; gastroesophageal cancer; histological slides; immune checkpoint; immunological status; innovation; malignant breast neoplasm; melanoma; multiplex assay; neoplastic cell; next generation; overtreatment; patient variability; personalized diagnostics; personalized health care; predictive marker; predictive test; preservation; programmed cell death ligand 1; protein expression; sample fixation; tissue fixing; tumor

Project Summary/Abstract Therapies against immunologic checkpoint proteins, such as PD-L1, have revolutionized the treatment of multiple malignancies, such as malignant melanoma, lung cancer, bladder cancer, or gastroesophageal cancer. All of these therapies depend on predictive biomarker assays for PD-L1 protein expression in tumor tissue to identify patients who will most likely respond. PD-L1 assays face two significant challenges: (i) they often need to be performed on small tissue biopsies where tumor tissue is limited and (ii) the threshold for positivity by immunohistochemistry (1%) is very small, highlighting crucial dependence on utmost precision and pre-analytical sample validity. To address the issue of small and limited samples in biopsies, we have developed a chromogenic multiplexed immunohistochemical assay that combines PD-L1 assessment with tissue and cell type-specific markers to provide a combined diagnostic and predictive assay on a single histologic slide. To move multiplexed assays around PD-L1 into the clinic, there exists an important knowledge gap: what are the patient-specific factors and specimen-related pre-analytical variables that can influence the readout of PD-L1 positivity? Very limited knowledge is available about these variables. Importantly, because the PD-L1 positivity threshold is so low and requires the reliable separation of two very small numbers (<1% vs. ≥1%), even minute pre-analytical variabilities would be expected to have significant negative impact on assay validity. In that respect, PD-L1 testing in tissue is particularly in need of extensive characterization and control of pre-analytical variability, even more so than other assays whose cut-off points lie in more favorable ranges. Our proposal is based on the hypothesis that both patient-specific factors (such as molecular features of the cancer, current immune status, prior drug therapy, etc.) and specimen-related factors (such as timing of biopsy, size of tissue, ischemic time, fixation protocol, etc.) can significantly influence subsequent biomarker measurements. We further hypothesize that solid knowledge about these influences will allow controlling for and mitigating patient-specific and specimen-related effects and will lead to more accurate and valid biomarker assessment. Aim 1: We will create a cohort to test the influence of patient-specific context factors. We will make use of our extensive immuno-oncology database and biobank of >5,000 patients. Aim 2: We will test how specimen-related pre-analytical variables affect the assay using a wide variety of fresh, frozen, and formalin fixed tissue types and sizes. Aim 3: Once we have defined optimal patient- specific and specimen-related procedures, we will validate our multiplex assay in a prospective cohort of immunotherapy patients at MSKCC. Significance: This project will yield abundant data about the pre-analytical variables that influence a PD-L1 multiplex immunohistochemistry assay. These data will inform optimal specimen acquisition and handling and strategies for avoiding or mitigating inaccurate assay results. Innovation: This will be the first study to systematically explore both patient context factors and specimen pre-analytics in multiplexed immunohistochemical testing for immuno-oncology. Our close collaboration with commercial test developers as part of our research team will accelerate the translation of our scientific findings into optimized assays for the direct benefit of patients.

项目摘要/摘要 针对免疫检查点蛋白的治疗，如PD-L1，已经彻底改变了多发性硬化症的治疗方法。 恶性肿瘤，如恶性黑色素瘤、肺癌、膀胱癌或胃食道癌。所有这些都是 治疗依赖于肿瘤组织中PD-L1蛋白表达的预测性生物标记物分析来识别 很可能会做出回应。PD-L1分析面临两个重大挑战：(1)它们经常需要在小组织上进行 在肿瘤组织有限且(Ii)免疫组织化学阳性阈值(1%)非常小的情况下进行活检， 强调对最高精密度和分析前样品有效性的关键依赖。 为了解决活组织检查中小样本和有限样本的问题，我们开发了一种显色多重 结合PD-L1检测与组织和细胞类型特异性标记物的免疫组织化学分析 一张组织切片上的联合诊断和预测分析。要将PD-L1周围的多路传输分析移到 临床上，存在着一个重要的知识鸿沟：哪些是患者特有的因素和标本相关的分析前 会影响PD-L1正性读数的变量？人们对这些变量的了解非常有限。 重要的是，因为PD-L1阳性阈值如此低，并且需要可靠地分离两个非常小的 数字(&lt；1%对≥1%)，即使是微小的分析前变异也会对 化验效度。在这方面，组织中的PD-L1检测特别需要广泛的表征和控制 分析前的可变性，甚至比其他分析的分界点位于更有利的范围内的分析更具可变性。 我们的建议是基于这样的假设，即患者特有的因素(例如癌症的分子特征， 当前免疫状态、既往药物治疗情况等)以及与标本相关的因素(如活检时间、组织大小、 缺血时间、固定方案等)会显著影响后续的生物标记物测量。我们进一步 假设对这些影响的扎实了解将允许控制和减轻患者特定的和 并将导致更准确和有效的生物标志物评估。目标1：我们将创建一个队列 以测试患者特定背景因素的影响。我们将利用我们广泛的免疫肿瘤学数据库 和5,000名患者的生物库。目标2：我们将测试与标本相关的分析前变量如何影响使用 各种新鲜的、冷冻的和福尔马林固定的组织类型和大小。目标3：一旦我们确定了最佳患者- 在特定的和标本相关的程序中，我们将在免疫治疗的前瞻性队列中验证我们的多重分析。 在MSKCC的患者。 意义：这个项目将产生大量关于影响PD-L1多路传输的分析前变量的数据 免疫组织化学检测。这些数据将为最佳样本获取和处理以及 避免或减轻不准确的化验结果。创新：这将是第一个系统地探索两者的研究 免疫肿瘤学多重免疫组织化学检测中的患者背景因素和标本预分析。 作为我们研究团队的一部分，我们与商业测试开发人员的密切合作将加速翻译 我们的科学发现转化为优化的分析方法，直接造福于患者。