Optical imaging of pancreas cancer organoids for drug development and personalized treatment

胰腺导管腺癌（PDAC）是一种侵袭性疾病， 所有癌症中预后最差（5年生存率为7%），在美国每年有超过40，000人死亡。标准- PDAC的护理外治疗包括手术和化疗。许多患者接受化疗后， 手术（“辅助治疗”），以改善无进展生存期。大约45-55%的患者存在 转移性疾病，如果耐受，立即开始化疗。不幸的是， 与化疗相关的毒性，并且没有工具来确定患者是否会从化疗中获益。 毒性更强的药物和毒性更弱的药物。此外，没有合理的系统将每个患者与 最有前途的抗癌新药最后，对于PDAC几乎没有有效的药物，因此， 加速药物开发的需求。因此，迫切需要改善PDAC患者的护理 通过降低毒性、合理的治疗计划和新药开发。 该提案的目标是开发新的细胞水平成像技术来预测治疗反应 在个体PDAC患者中，使用维持在3D培养物中的患者肿瘤的大悬液， （“类器官”）。这种非侵入性光学代谢成像（OMI）方法利用了内源性荧光 代谢辅酶NADH和FAD的强度和寿命，以在单细胞水平上成像药物反应 在完整的3D样本中的所有细胞中。这种单细胞分析允许异质性药物反应被 在肿瘤细胞和基质细胞的治疗时间过程中监测。这对于评估治疗很重要 在PDAC的高度异质性和基质微环境中的疗效。原发性肿瘤的应用 来自患者自身肿瘤的类器官也保持了细胞间的通讯，3D结构， 肿瘤-基质相互作用，这对准确评估药物反应至关重要。我们的初步数据显示， 原发性PDAC类器官中的OMI（1）可准确预测小鼠体内药物反应，（2）可识别药物 其有效地靶向肿瘤成纤维细胞用于基质重组和改善药物递送。我们还 在6名PDAC患者中进行人体试验的可行性。这个新的平台提供了巨大的潜力（1） 在相关患者样本上快速测试新药方案，从而加速PDAC药物开发，以及 (2)提供个性化的药物筛选，为每位患者确定最有效和毒性最小的治疗方法。 该提案将检验以下假设：原发性PDAC类器官的OMI可以准确预测体内 在小鼠和人类中的治疗功效。该方法将在PDAC小鼠模型上、在原发性 辅助治疗环境中的患者样本和转移性治疗中的主要患者样本 设置.建议开发动态单细胞评估技术，以监测药物反应， 完整的小鼠和人类PDAC类器官中的多种细胞类型为合理的药物治疗提供了巨大的希望。 开发和治疗计划，最终可以减少毒性和改善患者的结果。

Pancreatic ductal adenocarcinoma (PDAC) is an aggressive and poorly understood disease that has the worst prognosis of all cancers (5 year survival of 7%), with more than 40,000 deaths per year in the US. Standard- of-care treatment for PDAC includes surgery and chemotherapy. Many patients receive chemotherapy after surgery (“adjuvant treatment”), to improve progression-free survival. About 45-55% of patients present with metastatic disease and immediately begin chemotherapy if tolerated. Unfortunately, there are significant toxicities associated with chemotherapy, and there are no tools to determine whether a patient will benefit from a more toxic versus a less toxic drug. Additionally, there is no rational system to match each patient with the most promising new drug for their cancer. Finally, there are few effective drugs for PDAC, and therefore a demand for accelerated drug development. Thus, there is a critical need to improve the care of PDAC patients through reduced toxicities, rational treatment planning, and new drug development. The goal of this proposal is to develop novel cellular-level imaging technologies to predict treatment response in individual PDAC patients, using macro-suspensions of the patients’ tumors maintained in a 3D culture (“organoids”). This non-invasive optical metabolic imaging (OMI) approach exploits the intrinsic fluorescence intensity and lifetime of the metabolic co-enzymes NADH and FAD to image drug response on a single-cell level across all cells in the intact 3D sample. This single-cell analysis allows for heterogeneous drug response to be monitored over a treatment time-course in tumor cells and stromal cells. This is important for assessing treatment efficacy in the highly heterogeneous and stromal micro-environment of PDAC. The use of primary tumor organoids derived from the patients’ own tumor also maintains the cell-cell communication, 3D architecture, and tumor-stromal interactions that are critical to accurately assess drug response. Our preliminary data indicate that OMI in primary PDAC organoids (1) accurately predicts in vivo drug response in mice, and (2) can identify drugs that effectively target tumor fibroblasts for stromal re-organization and improved drug delivery. We have also established feasibility for human testing in 6 PDAC patients. This novel platform provides great potential for (1) rapidly testing new drug regimens on relevant patient samples, thus accelerating PDAC drug development, and (2) providing individualized drug screens to identify the most effective and least toxic treatment for each patient. This proposal will test the hypothesis that OMI of primary PDAC organoids can accurately predict in vivo treatment efficacy in mice and humans. This approach will be validated on mouse models of PDAC, on primary patient samples in the adjuvant treatment setting, and on primary patient samples in the metastatic treatment setting. The proposed development of dynamic, single-cell assessment techniques to monitor drug response in multiple cell types within intact mouse and human PDAC organoids holds great promise for rational drug development and treatment planning that could ultimately reduce toxicities and improve outcomes in patients.