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.

项目总结/摘要 针对免疫检查点蛋白（如PD-L1）的治疗已经彻底改变了多种肿瘤的治疗。 恶性肿瘤，如恶性黑色素瘤、肺癌、膀胱癌或胃食管癌。所有这些 治疗依赖于肿瘤组织中PD-L1蛋白表达的预测性生物标志物测定， 很可能会回应。PD-L1检测面临两个重大挑战：（i）它们通常需要在小组织上进行 活组织检查，其中肿瘤组织是有限的，和（ii）免疫组织化学的阳性阈值（1%）非常小， 强调了对最高精度和分析前样品有效性的关键依赖。 为了解决活检中小样本和有限样本的问题，我们开发了一种显色多重 免疫组化检测结合PD-L1评估与组织和细胞类型特异性标志物， 在单个组织学载玻片上进行联合诊断和预测测定。为了将PD-L1周围的多重检测转移到 在临床上，存在一个重要的知识差距：什么是患者的特定因素和药物相关的预分析 可能影响PD-L1阳性读数的变量？关于这些变量的知识非常有限。 重要的是，由于PD-L1阳性阈值非常低，需要可靠地分离两个非常小的 即使是微小的分析前变异性，预计也会对 测定有效性。在这方面，组织中的PD-L1检测特别需要广泛的表征和控制， 分析前的变异性，甚至比其他检测更好，其临界点位于更有利的范围内。 我们的建议是基于这样的假设，即患者特异性因素（如癌症的分子特征， 当前免疫状态、既往药物治疗等）以及与活检相关的因素（例如活检的时间，组织的大小， 缺血时间、固定方案等）可显著影响随后的生物标志物测量。我们进一步 假设关于这些影响的扎实知识将允许控制和减轻患者特异性 生物标志物相关的影响，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 和超过5,000名患者的生物库。目的2：我们将使用以下方法检测与分析前变量相关的干扰素如何影响测定 各种新鲜、冷冻和福尔马林固定的组织类型和尺寸。目标3：一旦我们确定了最佳患者- 特异性和免疫相关程序，我们将在一个前瞻性的免疫治疗队列中验证我们的多重检测 患者在MSKCC。 意义：本项目将产生关于影响PD-L1多重检测的分析前变量的丰富数据 免疫组织化学测定。这些数据将为最佳的标本采集和处理以及 避免或减轻不准确的测定结果。创新：这将是第一个系统地探讨这两个问题的研究。 患者背景因素和标本预分析在免疫肿瘤学的多重免疫组织化学检测。 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 将我们的科学发现转化为优化的检测方法，直接造福患者。