Wasson Elisa Marie, He Wei, Ahlquist Jesse, Hynes William Fredrick, Triplett Michael Gregory, Hinckley Aubree, Karelehto Eveliina, Gray-Sherr Delaney Ruth, Friedman Caleb Fisher, Robertson Claire, Shusteff Maxim, Warren Robert, Coleman Matthew A, Moya Monica Lizet, Wheeler Elizabeth K
Materials Engineering Division, Lawrence Livermore National Laboratory, Livermore, CA, United States.
Biosciences and Biotechnology Division, Lawrence Livermore National Laboratory, Livermore, CA, United States.
Front Bioeng Biotechnol. 2023 May 31;11:1193430. doi: 10.3389/fbioe.2023.1193430. eCollection 2023.
There is an urgent need to develop new therapies for colorectal cancer that has metastasized to the liver and, more fundamentally, to develop improved preclinical platforms of colorectal cancer liver metastases (CRCLM) to screen therapies for efficacy. To this end, we developed a multi-well perfusable bioreactor capable of monitoring CRCLM patient-derived organoid response to a chemotherapeutic gradient. CRCLM patient-derived organoids were cultured in the multi-well bioreactor for 7 days and the subsequently established gradient in 5-fluorouracil (5-FU) concentration resulted in a lower IC in the region near the perfusion channel versus the region far from the channel. We compared behaviour of organoids in this platform to two commonly used PDO culture models: organoids in media and organoids in a static (no perfusion) hydrogel. The bioreactor IC values were significantly higher than IC values for organoids cultured in media whereas only the IC for organoids far from the channel were significantly different than organoids cultured in the static hydrogel condition. Using finite element simulations, we showed that the total dose delivered, calculated using area under the curve (AUC) was similar between platforms, however normalized viability was lower for the organoid in media condition than in the static gel and bioreactor. Our results highlight the utility of our multi-well bioreactor for studying organoid response to chemical gradients and demonstrate that comparing drug response across these different platforms is nontrivial.
迫切需要开发针对已转移至肝脏的结直肠癌的新疗法，更重要的是，要开发改进的结直肠癌肝转移（CRCLM）临床前平台来筛选疗法的疗效。为此，我们开发了一种多孔可灌注生物反应器，能够监测CRCLM患者来源的类器官对化疗梯度的反应。将CRCLM患者来源的类器官在多孔生物反应器中培养7天，随后建立的5-氟尿嘧啶（5-FU）浓度梯度导致灌注通道附近区域的IC低于远离通道的区域。我们将该平台中类器官的行为与两种常用的PDO培养模型进行了比较：培养基中的类器官和静态（无灌注）水凝胶中的类器官。生物反应器的IC值显著高于在培养基中培养的类器官的IC值，而只有远离通道的类器官的IC与在静态水凝胶条件下培养的类器官有显著差异。使用有限元模拟，我们表明，使用曲线下面积（AUC）计算的总给药剂量在各平台之间相似，然而，培养基条件下类器官的归一化活力低于静态凝胶和生物反应器中的类器官。我们的结果突出了我们的多孔生物反应器在研究类器官对化学梯度反应方面的实用性，并表明在这些不同平台之间比较药物反应并非易事。
