A microfluidic-based PDAC organoid system reveals the impact of hypoxia in response to treatment

Abstract

Pancreatic Ductal Adenocarcinoma (PDAC) is estimated to become the second leading cause of cancer-related deaths by 2030 with mortality rates of up to 93%. Standard of care chemotherapeutic treatment only prolongs the survival of patients for a short timeframe. Therefore, it is important to understand events driving treatment failure in PDAC as well as identify potential more effective treatment opportunities. PDAC is characterized by a high-density stroma, high interstitial pressure and very low oxygen tension. The aim of this study was to establish a PDAC platform that supported the understanding of treatment response of PDAC organoids in mono-, and co-culture with pancreatic stellate cells (PSCs) under hypoxic and normoxic conditions. Cultures were exposed to Gemcitabine in combination with molecules targeting relevant molecular programs that could explain treatment specific responses under different oxygen pressure conditions. Two groups of treatment responses were identified, showing either a better effect in monoculture or co-culture. Moreover, treatment response also differed between normoxia and hypoxia. Modulation of response to Gemcitabine was also observed in presence of a Hypoxia-inducible factor (HIF) prolyl hydroxylase (PHD) inhibitor and HIF inhibitors. Altogether this highlights the importance of adjusting experimental conditions to include relevant oxygen levels in drug response studies in PDAC.

Understanding organotropism in cancer metastasis using microphysiological systems

“…to assess metastatic potential. 48 This hypoxia-driven angiogenic response not only supports tumor growth but also lays the groundwork for the metastatic cascade. The dynamics of the VEGF-induced tumor metastasis microenvironment were summarized in a multilayered blood vessel/tumor tissue chip.…”

Exploring New Dimensions of Tumor Heterogeneity: The Application of Single Cell Analysis to Organoid‐Based 3D In Vitro Models

“…Microenvironmental heterogeneity arising from dynamic gradients of oxygen and nutrients in the TME can also be modeled in microfluidic devices as they enable precise control over the system and can be integrated with sensors for real‐time monitoring. [ 128–138 ] For example, Dornhof et al. created a gas‐tight microfluidic device with two gel‐filled cell compartments surrounded by three parallel fluidic channels and integrated lactate, glucose and oxygen sensors that allowed establishment of hypoxia in the device by reducing media perfusion or performing asymmetric perfusion through a side channel.…”

“…Using this model, they explored the combined effect of hypoxia and PSCs on the PDO response to gemcitabine and other therapeutic agents. [ 129 ] Accordingly, integration of hypoxia, stromal cells and PDOs using microfluidic devices as a platform may offer a promising opportunity for future investigations of the TME.…”

“…Microenvironmental heterogeneity arising from dynamic gradients of oxygen and nutrients in the TME can also be modeled in microfluidic devices as they enable precise control over the system and can be integrated with sensors for real-time monitoring. [128][129][130][131][132][133][134][135][136][137][138] For example, Dornhof et al created a gastight microfluidic device with two gel-filled cell compartments surrounded by three parallel fluidic channels and integrated lactate, glucose and oxygen sensors that allowed establishment of hypoxia in the device by reducing media perfusion or performing asymmetric perfusion through a side channel. They showed that breast cancer stem cell-derived spheroids cultured in this device were responsive to environmental conditions and had cancerspecific metabolic characteristics, such as the production of high amounts of lactate despite normoxic conditions.…”

“…[128] Other microfluidic devices predominantly tested with tumor cell lines and tumor spheroids have used diffusion barriers and gas control chambers to establish hypoxia, often in conjunction with hypoxia tracking probes to visualize tumor cells experiencing hypoxia. [129][130][131][132][133][134][135][136][137] In one example with PDOs, Geyer et al used the commercially available OrganoPlate 2-Lane-a plate-based microfluidic device with a cell-gel suspension channel and a parallel perfusion channel with gravity-driven flow-in a hypoxia chamber to create a hypoxic PDAC PDO and PSC co-culture model. Using this model, they explored the combined effect of hypoxia and PSCs on the PDO response to gemcitabine and other therapeutic agents.…”