Department of Surgical Oncology.
Department of Experimental Therapeutics.
J Clin Invest. 2022 Dec 15;132(24):e151604. doi: 10.1172/JCI151604.
BACKGROUNDPancreatic ductal adenocarcinoma (PDAC) is one of the most lethal malignancies, with unpredictable responses to chemotherapy. Approaches to assay patient tumors before treatment and identify effective treatment regimens based on tumor sensitivities are lacking. We developed an organoid-based platform (OBP) to visually quantify patient-derived organoid (PDO) responses to drug treatments and associated tumor-stroma modulation for personalized PDAC therapy.METHODSWe retrospectively quantified apoptotic responses and tumor-stroma cell proportions in PDOs via 3D immunofluorescence imaging through annexin A5, α-smooth muscle actin (α-SMA), and cytokeratin 19 (CK-19) levels. Simultaneously, an ex vivo organoid drug sensitivity assay (ODSA) was used to measure responses to standard-of-care regimens. Differences between ODSA results and patient tumor responses were assessed by exact McNemar's test.RESULTSImmunofluorescence signals, organoid growth curves, and Ki-67 levels were measured and authenticated through the OBP for up to 14 days. ODSA drug responses were not different from patient tumor responses, as reflected by CA19-9 reductions following neoadjuvant chemotherapy (P = 0.99). PDOs demonstrated unique apoptotic and tumor-stroma modulation profiles (P < 0.0001). α-SMA/CK-19 ratio levels of more than 1.0 were associated with improved outcomes (P = 0.0179) and longer parental patient survival by Kaplan-Meier analysis (P = 0.0046).CONCLUSIONHeterogenous apoptotic drug responses and tumor-stroma modulation are present in PDOs after standard-of-care chemotherapy. Ratios of α-SMA and CK-19 levels in PDOs are associated with patient survival, and the OBP could aid in the selection of personalized therapies to improve the efficacy of systemic therapy in patients with PDAC.FUNDINGNIH/National Cancer Institute grants (K08CA218690, P01 CA117969, R50 CA243707-01A1, U54CA224065), the Skip Viragh Foundation, the Bettie Willerson Driver Cancer Research Fund, and a Cancer Center Support Grant for the Flow Cytometry and Cellular Imaging Core Facility (P30CA16672).
背景
胰腺导管腺癌 (PDAC) 是最致命的恶性肿瘤之一，对化疗的反应不可预测。在治疗前对患者肿瘤进行检测，并根据肿瘤敏感性确定有效的治疗方案的方法尚未开发。我们开发了一种基于类器官的平台 (OBP)，用于通过 3D 免疫荧光成像来可视化定量患者来源的类器官 (PDO) 对药物治疗的反应以及相关的肿瘤-基质调节，以实现个性化 PDAC 治疗。
方法
我们通过 Annexin A5、α-平滑肌肌动蛋白 (α-SMA) 和细胞角蛋白 19 (CK-19) 水平的 3D 免疫荧光成像，回顾性地定量了 PDO 中的细胞凋亡反应和肿瘤-基质细胞比例。同时，使用离体类器官药物敏感性测定 (ODSA) 来测量对标准治疗方案的反应。通过确切的 McNemar 检验评估 ODSA 结果与患者肿瘤反应之间的差异。
结果
通过 OBP 测量和验证了多达 14 天的免疫荧光信号、类器官生长曲线和 Ki-67 水平。ODSA 药物反应与患者肿瘤反应没有差异，如新辅助化疗后 CA19-9 降低所反映的那样（P=0.99）。PDO 表现出独特的细胞凋亡和肿瘤-基质调节特征（P<0.0001）。α-SMA/CK-19 比值大于 1.0 与改善的结果相关（P=0.0179），并通过 Kaplan-Meier 分析延长了亲代患者的生存时间（P=0.0046）。
结论
在标准治疗化疗后，PDO 中存在异质性的细胞凋亡药物反应和肿瘤-基质调节。PDO 中α-SMA 和 CK-19 水平的比值与患者生存相关，OBP 可以帮助选择个性化治疗方法，以提高 PDAC 患者全身治疗的疗效。
资助
NIH/美国国立癌症研究所拨款（K08CA218690、P01CA117969、R50CA243707-01A1、U54CA224065）、Skip Viragh 基金会、Bettie Willerson Driver 癌症研究基金以及流式细胞术和细胞成像核心设施的癌症中心支持拨款（P30CA16672）。
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