|
Abstract INTRODUCTION:
Ovarian cancer is the deadliest gynecologic malignancy, with high-grade serous carcinoma (HGSC) accounting for ~70% of cases. HGSC originates in the fallopian tube epithelium and metastasizes to the ovaries and omentum. We have found that epinephrine enhances lipid release from omental adipocytes, creating a pro-metastatic microenvironment. Using mass spectrometry imaging (MSI) of 3D co-cultures of HGSC cells, murine omental tissues, and adipocyte spheroids, alongside LC-MS(/MS) lipidomic profiling and invasion assays, we investigated this metabolic crosstalk. Our findings suggest that epinephrine acts as an autocrine signal in the tumor microenvironment to enhance lipid release, fostering HGSC invasion and disease progression. Understanding these interactions offers insight into tumor-microenvironment dynamics and could guide the development of targeted therapies to inhibit HGSC metastasis.
METHODS:
A 3D co-culture system in agarose was optimized to study interactions between murine omental tissues, adipocytes, and murine oviductal epithelial (MOE) cells. MALDI MSI was performed using a Bruker timsTOF flex mass spectrometer with a 50-micron spatial resolution in positive reflectron mode. A 50:50 CHCA:DHB matrix was applied using an HTX TMSprayer, enabling prioritization of lipid signals intensified in cancer-omentum/adipocyte interactions.
For lipidomic profiling, LC-MS(/MS) analysis was performed in reverse-phase using both positive and negative ionization modes to identify differentially expressed lipids in adipocyte-conditioned media treated with a β-adrenergic receptor agonist (epinephrine) or antagonist (propranolol). These conditioned media were also used to evaluate cell behaviors, including proliferation, migration, and invasion.
RESULTS:
MSI data revealed that epinephrine is produced in 3D co-cultures involving HGSC cells, omental tissues, and adipocyte spheroids. Epinephrine-treated adipocyte-conditioned media significantly enhanced HGSC cell invasion, demonstrating the pro-metastatic effects of β-adrenergic signaling. Conversely, propranolol, a β-adrenergic receptor antagonist, effectively inhibited these effects by blocking endogenous epinephrine action.
After optimizing lipid extraction strategies for adipocyte-conditioned media, LC-MS/MS data acquired in both ionization modes were analyzed to classify lipid subclasses and their relative abundance across treatments. Visualization tools such as Volcano Plots and Molecular Networks were used to interpret the data, leveraging Cytoscape for network analysis and LinexWorkflow for lipid metabolic pathway mapping.
Lipidomic profiling revealed an increased release of specific lipid classes under epinephrine treatment, particularly glycerolipids and sterols, such as MG O-6:0 and ST 26:6. These findings suggest that β-adrenergic receptor activation enhances omental lipolysis, potentially contributing to metabolic reprogramming within the tumor microenvironment. This reprogramming may create a lipid-rich niche that supports HGSC progression.
Furthermore, MALDI MSI provided spatially resolved confirmation of lipid accumulation at tumor-omentum interfaces within the 3D co-culture system. These localized lipid enrichments further support the hypothesis that metabolic crosstalk mediated by β-adrenergic signaling plays a pivotal role in facilitating tumor invasion and metastasis.
CONCLUSION:
Our findings highlight the role of epinephrine in modulating lipid dynamics within the HGSC tumor microenvironment. By integrating MALDI MSI and lipidomics, we reveal how β-adrenergic signaling enhances omental lipolysis, creating a pro-metastatic lipid-rich niche. These insights could guide the development of therapeutic strategies targeting metabolic vulnerabilities in HGSC.
|
= Discovery stage. (53.14%, 2025)
= Translation stage. (22.33%, 2025)
= Clinically available. (24.53%, 2025)
