Armstrong Andrew, Haque Muhammad R, Mirbagheri Sina, Barlass Usman, Gilbert Douglas Z, Amin Jaimin, Singh Ajaypal, Naqib Ankur, Bishehsari Faraz
Department of Internal Medicine, Division of Gastroenterology, Rush University Medical Center, Chicago, IL 60612, USA.
Division of Digestive Diseases, Rush Center for Integrated Microbiome & Chronobiology Research, Rush University Medical Center, Chicago, IL 60612, USA.
Biomedicines. 2021 Jun 23;9(7):705. doi: 10.3390/biomedicines9070705.
Pancreatic ductal adenocarcinoma (PDA) is an extremely lethal malignancy arising from the pancreas. The treatment of PDA is complicated by ineffective treatments and a lack of biomarkers predictive of treatment success. We have designed a patient-derived organoid (PDO) based high-throughput drug screening assay to model treatment response to a variety of conventional and investigational treatments for PDA. Consecutive patients undergoing endoscopic ultrasound-guided fine-needle biopsy for tissue diagnosis of PDA at Rush University Medical Center were offered to participate in the study. Biopsies were immediately processed to develop organoids. Fifteen PDOs were screened for sensitivity to 18 compounds, including conventional PDA chemotherapies and FDA-approved investigational targeted therapies in cancer using Cell-titer GLO 3D (Promega) cell viability assay. The area under the curve (AUC) was calculated and normalized to the maximum area under the curve to generate a normalized AUC between 0 and 1. Molecular profiling of PDOs was conducted using RNA-seq. Human PDA transcriptomic was extracted from The Cancer Genome Atlas (TCGA). The drug response curves were reproducible. We observed variation in response to conventional therapies overall as well as among individual patients. There were distinct transcriptome signatures associated with response to the conventional chemotherapeutics in PDA. The transcriptomic profile of overall resistance to conventional therapies in our study was associated with poor survival in PDA patients in TCGA. Our pathway analysis for targeted drugs revealed a number of predictors of response associated with the mechanism of action of the tested drug. The multiplex organoid-based drug assay could be used in preclinical to inform patient stratification and therapeutic selection in PDA. When combined with omics data, ex vivo response to treatment could help identify gene signatures associated with response to novel therapies.
胰腺导管腺癌（PDA）是一种起源于胰腺的极具致死性的恶性肿瘤。PDA的治疗因治疗效果不佳以及缺乏预测治疗成功的生物标志物而变得复杂。我们设计了一种基于患者来源类器官（PDO）的高通量药物筛选试验，以模拟对PDA的多种传统和研究性治疗的反应。连续在拉什大学医学中心接受内镜超声引导下细针穿刺活检以进行PDA组织诊断的患者被邀请参与该研究。活检组织立即进行处理以培养类器官。使用Cell-titer GLO 3D（普洛麦格公司）细胞活力测定法，对15个PDO进行了对18种化合物的敏感性筛选，这些化合物包括传统的PDA化疗药物以及美国食品药品监督管理局批准的癌症研究性靶向治疗药物。计算曲线下面积（AUC）并将其归一化至最大曲线下面积，以生成0到1之间的归一化AUC。使用RNA测序对PDO进行分子谱分析。从癌症基因组图谱（TCGA）中提取人类PDA转录组数据。药物反应曲线具有可重复性。我们观察到总体上以及个体患者之间对传统疗法的反应存在差异。在PDA中，存在与对传统化疗药物的反应相关的独特转录组特征。在我们的研究中，对传统疗法总体耐药的转录组特征与TCGA中PDA患者的不良生存相关。我们对靶向药物的通路分析揭示了一些与受试药物作用机制相关的反应预测因子。基于多类器官的药物试验可用于临床前研究，为PDA患者的分层和治疗选择提供信息。当与组学数据相结合时，体外治疗反应有助于识别与对新疗法的反应相关的基因特征。
