Complement activation at the interface between adipocytes and cancer cells drives tumor progression
Andres Valdivia, Ana Maria Isac, Horacio Cardenas, Guangyuan Zhao, Yaqi Zhang, Hao Huang, Jian-Jun Wei, Mauricio Cuello-Fredes, Sumie Kato, Fernán Gómez-Valenzuela, Francoise Gourronc, Aloysius Klingelhutz, Daniela Matei
Andres Valdivia, Ana Maria Isac, Horacio Cardenas, Guangyuan Zhao, Yaqi Zhang, Hao Huang, Jian-Jun Wei, Mauricio Cuello-Fredes, Sumie Kato, Fernán Gómez-Valenzuela, Francoise Gourronc, Aloysius Klingelhutz, Daniela Matei
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Research Article Cell biology Oncology

Complement activation at the interface between adipocytes and cancer cells drives tumor progression

Abstract

The omentum is the primary site of metastasis for ovarian cancer (OC). Interactions between cancer cells and adipocytes drive an invasive and prometastatic phenotype. Here we studied cancer cell–adipocyte crosstalk by using a direct coculture model with immortalized human visceral nondiabetic pre-adipocytes (VNPADs) and OC cells. We demonstrated increased proliferation, invasiveness, and resistance to cisplatin of cocultured compared with monocultured OC cells. RNA sequencing of OC cells from coculture versus monoculture revealed significant transcriptomic changes, identifying over 200 differentially expressed genes common to OVCAR5 and OVCAR8 cell lines. Enriched pathways included PI3K/AKT and complement activation. Lipid transfer into OC cells from adipocytes induced upregulation of complement C3 and C5 proteins. Inhibiting C3 or C5 reversed the invasive phenotype and C3 knockdown reduced tumor progression in vivo. Increased C3 expression was observed in omental implants compared with primary ovarian tumors and C3 secretion was higher in OC ascites from high-BMI versus low-BMI patients. C3 upregulation in OC cells involved activation of the ATF4-mediated integrated stress response (ISR). Overall, adipocyte–cancer cell interactions promoted invasiveness and tumorigenesis via lipid transfer, activating the ISR, and upregulating complement proteins C3 and C5.

Authors

Andres Valdivia, Ana Maria Isac, Horacio Cardenas, Guangyuan Zhao, Yaqi Zhang, Hao Huang, Jian-Jun Wei, Mauricio Cuello-Fredes, Sumie Kato, Fernán Gómez-Valenzuela, Francoise Gourronc, Aloysius Klingelhutz, Daniela Matei

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Figure 3

C3 and C5 are overexpressed in OC cells after coculture with adipocytes.

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C3 and C5 are overexpressed in OC cells after coculture with adipocytes....
(A) mRNA expression levels of C3 and C5 measured by qRT-PCR in monocultured and cocultured OVCAR8 and OVCAR5 cells (mean ± SD, n = 3). (B) ELISA-measured protein levels of C3 and C5 secreted in conditioned media of monocultured (Ctrl) and cocultured (Adipo CC) OVCAR8 and OVCAR5 cells (mean ± SD, n = 5). Base VNPAD media and adipocyte-conditioned media (Adipo CM) were included as additional controls. (C) Proliferation (CCK8 assay) of OVCAR8 (left panel) and OVCAR5 (right panel) cells maintained in monoculture versus cocultured with adipocytes and treated with 240 nM C3R/C5R inhibitor SB290157. (D) Proliferation assay of shCtrl and shC3aR1 (clones A and B) OVCAR5 cells cultured in base medium (Ctrl) versus adipocyte-conditioned media (AdipoCM); n = 4 replicates. (E) Proliferation (CCK8 assay) of shCtrl and shC3 OVCAR5 cells maintained as monoculture (Ctrl) versus cocultured with adipocytes (AdipoCC) (mean ± SD, n = 3). (F) Western blotting for C3, AKT, and p-Ser473 AKT pathway in shCtrl and shC3D OVCAR5 cells grown as monoculture or cocultured with adipocytes. (G) Representative picture of colony formation assay of OVCAR5 cells shCtrl and shC3. (H) Numbers of colonies (mean ± SD, n = 3) determined in a colony formation assay with shCtrl and shC3 OVCAR5 cells. (I) Proliferation (CCK8 assay) of shCTRL and shC3 (clone D) OVCAR5 cells cultured either in base medium (Ctrl) or treated with 50 ng or 100 ng recombinant C3. (J) qRT-PCR measurements (mean ± SD, n = 2) of C3 mRNA in primary cells derived from human HGSOC tumors (n = 2 tumors, nos. 148 and 154) and grown as monoculture or cocultured with adipocytes. (K) Western blotting for C3 in the primary OC cells described in panel I. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001 by 1-way ANOVA with Tukey’s post hoc test (A, B, and H) or 2-way ANOVA with Tukey’s post hoc test (C, D, E, and I).