PO.CL01.06 · 临床研究
为多中心临床试验实施一种新型免疫监测策略，以实现澳大利亚偏远地区个体癌症预后的公平性
Implementation of a novel immune monitoring strategy for multicenter clinical trials to enable equity in cancer prognosis for individuals from remote settings of Australia
作者与单位 Authors & Affiliations
摘要 Abstract
中文摘要
改善癌症预后始于从临床试验中收集高质量数据。然而，由于可能影响数据质量的后勤挑战，通过流式细胞术执行纵向免疫表型研究十分复杂。当考虑到澳大利亚偏远和农村社区时，免疫监测的挑战更加严峻，这些社区既承受着更高的癌症患病率，又面临更差的预后。矛盾的是，这些社区在临床试验中往往代表性不足。我们团队开发了一种新型的基于血液的"免疫特征"，能够稳健地预测黑色素瘤和肺癌中对靶向PD-1/PD-L1通路的检查点治疗产生临床应答失败。我们现已将这些开创性发现付诸偏远临床环境的实施，并配合一个扩展的CyTOF™检测panel，用于对癌症患者外周血样本进行50多个参数的分析。CyTOF技术的最新进展促成了异步样本采集和染色的高度简化工作流程，与当前医院实验室（通常按不可预测的时间表运行）兼容，并可延伸至资源匮乏、临床试验基础设施有限的偏远环境。这一独特优势通过使用稳定的、冻干的CyTOF抗体混合物和易于遵循的方案实现，该方案在最大限度减少所需样本量的同时产生可重复的染色。此外，这一独特的质谱流式细胞术工作流程通过样本条形码化和染色样本冷冻运往中心站点进行批量采集，提供了灵活性并最大限度减少了技术变异。因此，本研究旨在展示大型panel的临床影响以及CyTOF技术在多站点临床试验中高度简化且稳健工作流程方面的实用性。在此我们展示通过在澳大利亚三个站点、多种临床实施设置下开展的100份样本试点研究，实施这一新型工作流程的发现。开展了偏远地区异步样本采集，以比较两种表面染色方法：i）PBMC与ii）去血浆全血，分别适用于易于或不易获得离心条件的环境。染色样本运往中心实验室进行处理。对于冷冻保存的全血，首先进行基于微珠的粒细胞清除，随后所有样本一起进行多重条形码化，用于关键功能标志物的胞内抗体染色。在样本采集站点和染色方法之间实现了分析协调统一。总体而言，这一工作流程使得能够接触到服务不足的社区，首次促进了免疫表型研究的公平性和可扩展性，以利用真正地理分散的临床中心。
查看英文原文 English abstract
Improved cancer prognosis begins with the collection of high-quality data from clinical trials. However, execution of longitudinal immune phenotyping studies by flow cytometry is complex due to logistical challenges that could affect data quality. The challenges of immune monitoring are exacerbated when considering remote and rural communities in Australia, which are burdened by both an increased prevalence and worse prognosis of cancer. Paradoxically, these communities are often underrepresented in clinical trials. Our group has developed a novel blood-based “immune signature” that robustly predicts failure to make a clinical response to checkpoint therapies targeting the PD-1/PD-L1 pathway in melanoma and lung cancer. We have now taken these groundbreaking findings to implement in remote clinical settings, along with an expanded CyTOF™ panel for 50-plus-parameter analysis of cancer patient peripheral blood samples. Recent developments in CyTOF technology facilitate a highly simplified workflow of asynchronous sample collection and staining, compatible with current hospital laboratories, which typically run on unpredictable schedules, to remote settings, which are resource-poor and have limited clinical trial infrastructure. This is uniquely enabled by using a stable, dried-down cocktail of CyTOF antibodies and an easy-to-follow protocol that yields reproducible staining while minimizing sample required. Furthermore, this unique mass cytometry workflow provides flexibility and minimizes technical variation via sample barcoding and freezing of stained samples for shipment to a central site for batch acquisition. As such, this study is designed to demonstrate both clinical impact of the large panel and utility of CyTOF technology for a highly simplified and robust workflow for multi-site clinical trials. Here we present findings from the implementation of this novel workflow through a 100-sample pilot study at three sites across Australia, in a variety of clinical implementation setups. Remote asynchronous sample collection was performed to compare two surface staining approaches on i) PBMC versus ii) plasma-depleted whole blood to suit settings where centrifugation access was readily available or not, respectively. Stained samples were shipped to a central lab for processing. In the case of cryopreserved whole blood, bead-based granulocyte depletion was first performed, with all samples subsequently multiplex barcoded together for intracellular antibody staining of key functional markers. Analysis harmonization was achieved across sample collection sites and staining approaches. Overall, this workflow enables access to underserved communities, facilitating for the first time equity and scalability of immune phenotyping studies to harness truly geographically dispersed clinical centers.
利益披露 Disclosure
L. J. Tracey, None..
N. Pulyani, None..
R. Auzins, None..
H. McGuire, None.