Gillette Amani A, Babiarz Christopher P, VanDommelen Ava R, Pasch Cheri A, Clipson Linda, Matkowskyj Kristina A, Deming Dustin A, Skala Melissa C
Department of Biomedical Engineering, University of Wisconsin, Madison, WI 53706, USA.
Department of Medicine, Division of Hematology, Oncology and Palliative Care, School of Medicine and Public Health, University of Wisconsin, Madison, WI 53705, USA.
Cancers (Basel). 2021 Apr 14;13(8):1873. doi: 10.3390/cancers13081873.
Gastroenteropancreatic neuroendocrine tumors (GEP-NET) account for roughly 60% of all neuroendocrine tumors. Low/intermediate grade human GEP-NETs have relatively low proliferation rates that animal models and cell lines fail to recapitulate. Short-term patient-derived cancer organoids (PDCOs) are a 3D model system that holds great promise for recapitulating well-differentiated human GEP-NETs. However, traditional measurements of drug response (i.e., growth, proliferation) are not effective in GEP-NET PDCOs due to the small volume of tissue and low proliferation rates that are characteristic of the disease. Here, we test a label-free, non-destructive optical metabolic imaging (OMI) method to measure drug response in live GEP-NET PDCOs. OMI captures the fluorescence lifetime and intensity of endogenous metabolic cofactors NAD(P)H and FAD. OMI has previously provided accurate predictions of drug response on a single cell level in other cancer types, but this is the first study to apply OMI to GEP-NETs. OMI tested the response to novel drug combination on GEP-NET PDCOs, specifically ABT263 (navitoclax), a Bcl-2 family inhibitor, and everolimus, a standard GEP-NET treatment that inhibits mTOR. Treatment response to ABT263, everolimus, and the combination were tested in GEP-NET PDCO lines derived from seven patients, using two-photon OMI. OMI measured a response to the combination treatment in 5 PDCO lines, at 72 h post-treatment. In one of the non-responsive PDCO lines, heterogeneous response was identified with two distinct subpopulations of cell metabolism. Overall, this work shows that OMI provides single-cell metabolic measurements of drug response in PDCOs to guide drug development for GEP-NET patients.
胃肠胰神经内分泌肿瘤（GEP-NET）约占所有神经内分泌肿瘤的60%。低/中级别人类GEP-NET的增殖率相对较低，动物模型和细胞系无法重现这一特征。短期患者来源的癌症类器官（PDCO）是一种三维模型系统，有望重现高分化的人类GEP-NET。然而，由于GEP-NET PDCO组织体积小且增殖率低这一疾病特征，传统的药物反应测量方法（即生长、增殖）在GEP-NET PDCO中并不有效。在此，我们测试了一种无标记、非破坏性的光学代谢成像（OMI）方法，以测量活的GEP-NET PDCO中的药物反应。OMI可捕获内源性代谢辅因子NAD(P)H和FAD的荧光寿命和强度。OMI此前已在其他癌症类型的单细胞水平上提供了准确的药物反应预测，但这是首次将OMI应用于GEP-NET的研究。OMI测试了GEP-NET PDCO对新型药物组合的反应，具体为ABT263（维托克洛司），一种Bcl-2家族抑制剂，以及依维莫司，一种抑制mTOR的标准GEP-NET治疗药物。使用双光子OMI在源自7名患者的GEP-NET PDCO系中测试了对ABT263、依维莫司及其组合的治疗反应。OMI在治疗后72小时测量了5个PDCO系对联合治疗的反应。在一个无反应的PDCO系中，通过两个不同的细胞代谢亚群鉴定出异质性反应。总体而言，这项工作表明，OMI可提供PDCO中药物反应的单细胞代谢测量，以指导GEP-NET患者的药物开发。
