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The microenvironment induces collective migration in SDHB-silenced mouse pheochromocytoma spheroids
Vanessa D'Antongiovanni1, Serena Martinelli1, Susan Richter2
1Department of Experimental and Clinical Biomedical SciencesUniversity of Florence, Florence, Italy.
Abstract:
Pheochromocytomas (Pheos) and paragangliomas (PGLs) are neuroendocrine tumors. Approximately 30-40% of Pheos/PGLs are due to germline mutations in one of the susceptibility genes, including those encoding the succinate dehydrogenase subunits A-D (SDHA-D). Up to 2/3 of patients affected by SDHB mutated Pheo/PGL develop metastatic disease with no successful cure at present. Here, for the first time, we evaluated the effects of SDHB silencing in a three dimension (3D) culture using spheroids of a mouse Pheo cell line silenced or not (wild type/wt/control) for the SDHB subunit. We investigated the role of the microenvironment on spheroid growth and migration/invasion by co-culturing SDHB-silenced or wt spheroids with primary cancer-activated fibroblasts (CAFs). When spheroids were co-cultured with fibroblasts, SDHB-silenced cells showed a significant increase in matrigel invasion as demonstrated by the computation of the migratory areas (P < 0.001). Moreover, cells detaching from the SDHB-silenced spheroids moved collectively, unlike the cells of wt spheroids that moved individually. Additionally, SDHB-silenced spheroids developed long filamentous formations along which clusters of cells migrated far away from the spheroid, whereas these structures were not present in wt spheroids. We found that lactate, largely secreted by CAFs, plays a specific role in promoting migration only of SDHB-silenced cells. In this study, we demonstrated that SDHB silencing per se increases tumor cell migration/invasion and that microenvironment, as represented by CAFs, plays a pivotal role in enhancing collective migration/invasion in Pheo SDHB-silenced tumor cells, suggesting their role in increasing the tumor metastasizing potential.
Insights
Pheochromocytoma and paraganglioma cells with silenced SDHB showed increased invasion and collective migration. Cancer-associated fibroblasts and lactate further enhanced this metastatic potential in 3D cultures.
Area of Science:
- Endocrinology
- Oncology
- Cancer Biology
Background:
- Pheochromocytomas (Pheos) and paragangliomas (PGLs) are neuroendocrine tumors often linked to germline mutations.
- Mutations in succinate dehydrogenase subunit B (SDHB) are associated with aggressive disease and metastasis in Pheo/PGL patients.
- Current treatments for metastatic Pheo/PGL, particularly those with SDHB mutations, are limited.
Purpose of the Study:
- To investigate the impact of SDHB gene silencing on Pheo cell behavior in a 3D culture model.
- To explore the role of the tumor microenvironment, specifically cancer-activated fibroblasts (CAFs), in modulating the migratory and invasive properties of SDHB-silenced Pheo cells.
- To identify potential mechanisms, such as lactate production, contributing to enhanced tumor cell invasion.
Main Methods:
- Utilized a 3D spheroid culture system with a mouse Pheo cell line, genetically modified to silence the SDHB subunit.
- Co-cultured SDHB-silenced and wild-type (wt) spheroids with primary cancer-activated fibroblasts (CAFs).
- Quantified spheroid invasion using matrigel assays and analyzed cell migration patterns (collective vs. individual) and spheroid morphology.
Main Results:
- SDHB silencing significantly increased matrigel invasion of Pheo spheroids (P < 0.001).
- SDHB-silenced cells exhibited collective migration and formed filamentous structures, unlike wt cells which migrated individually.
- Co-culture with CAFs enhanced invasion, and lactate secreted by CAFs specifically promoted migration of SDHB-silenced cells.
Conclusions:
- SDHB silencing intrinsically enhances tumor cell migration and invasion in Pheochromocytoma/Paraganglioma.
- The tumor microenvironment, particularly CAFs and their secreted lactate, plays a crucial role in promoting collective migration and invasion of SDHB-silenced Pheo/PGL cells.
- These findings highlight the potential of targeting the microenvironment to inhibit metastasis in SDHB-mutated Pheo/PGL.
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