Targeting the Formyl Peptide Receptor type 1 to prevent the adhesion of ovarian cancer cells onto mesothelium and subsequent invasion

Abstract

Background: The biological behavior of epithelial ovarian cancer (EOC) is unique since EOC cells metastasize early to the peritoneum. Thereby, new anti-target agents designed to block trans-coelomic dissemination of EOC cells may be useful as anti-metastatic drugs. The Urokinase Plasminogen Activator Receptor (uPAR) is overexpressed in EOC tissues, and its truncated forms released in sera and/or ascitic fluid are associated with poor prognosis and unfavorable clinical outcome. We documented that uPAR triggers intra-abdominal dissemination of EOC cells through the interaction of its 84-95 sequence with the Formyl Peptide Receptor type 1 (FPR1), even as short linear peptide Ser-Arg-Ser-Arg-Tyr (SRSRY). While the pro-metastatic role of uPAR is well documented, little information regarding the expression and role of FPR1 in EOC is currently available. Methods: Expression levels of uPAR and FPR1 in EOC cells and tissues were assessed by immunofluorescence, Western blot, or immunohystochemistry. Cell adhesion to extra-cellular matrix proteins and mesothelium as well as mesothelium invasion kinetics by EOC cells were monitored using the xCELLigence technology or assessed by measuring cell-associated fluorescence. Cell internalization of FPR1 was identified on multiple z-series by confocal microscopy. Data from in vitro assays were analysed by one-way ANOVA and post-hoc Dunnett t-test for multiple comparisons. Tissue microarray data were analyzed with the Pearson's Chi-square (χ 2) test. Results: Co-expression of uPAR and FPR1 by SKOV-3 and primary EOC cells confers a marked adhesion to vitronectin. The extent of cell adhesion decreases to basal level by pre-exposure to anti-uPAR84-95 Abs, or to the RI-3 peptide, blocking the uPAR84-95/FPR1 interaction. Furthermore, EOC cells exposed to RI-3 or desensitized with an excess of SRSRY, fail to adhere also to mesothelial cell monolayers, losing the ability to cross them. Finally, primary and metastatic EOC tissues express a high level of FPR1. Conclusions: Our findings identify for the first time FPR1 as a potential biomarker of aggressive EOC and suggests that inhibitors of the uPAR84-95/FPR1 crosstalk may be useful for the treatment of metastatic EOC.

Extracellular Matrix: A Treasure Trove in Ovarian Cancer Dissemination and Chemotherapeutic Resistance

“…The molecular mechanism underlying these processes may be targeted to prevent tumor spread. Figure 2 shows a schematic view of the pathophysiology of ovarian cancer metastasis [ 4 , 5 , 7 , 9 , 12 - 14 , 16 , 18 - 23 , 26 - 28 ].…”

“…Urokinase plasminogen activator receptor (uPAR) interacts with formyl peptide receptor 1 (FPR1) and promotes tumor cell adhesion to mesothelial cells of peritoneum and vitronectin. Higher expression of uPAR and FPR1 was correlated with metastasis and poor clinical outcome [ 14 ].…”

“…Zhao et al uncovered the role of an angiotensin receptor blocker (AT-1 receptor blocker) Losartan in decreasing the proliferation of ECM protein collagen, thereby leading to lesser drug resistance and decreased ascites in patients [ 6 ]. Several pathways (Janus kinase and signal transducer and activator of transcription proteins JAK/STAT3, mitogen-activated protein kinase-7 MAPK-7) [ 7 , 8 ], cell surface receptors like integrins [ 9 ], proteins such as microfibril-associated protein-5 (MFAP-5) [ 10 ], Tau protein [ 11 ], collagen triple helix repeat containing-1 (CTHRC-1) [ 12 ], receptors such as discoidin domain receptor-2 (DDR-2) [ 13 ], formyl peptide receptor type 1 (FPR1) [ 14 ], cytokines like transforming growth factor-β (TGF-β) [ 15 ], enzymes such as lysyl oxidase (LOX) [ 4 ], integrin-linked kinase (ILK) [ 16 ], and histone deacetylase-4 (HDAC-4) have been well characterized in the literature [ 17 ]. Glucocorticoids have also been shown to be associated with tumor cell adhesion and resistance to chemotherapeutic agents [ 18 ].…”

Breast Tumor Cell Invasion and Pro-Invasive Activity of Cancer-Associated Fibroblasts Co-Targeted by Novel Urokinase-Derived Decapeptides

“…Fibroblasts were fixed in 4% paraformaldehyde (PFA), permeabilized with 0.1% Triton X-100 and incubated with anti-α-SMA overnight at 4 • C. Then, cells were stained with secondary goat anti-rabbit IgG H&L AlexaFluor-488 (A-21204, Life Technologies, Carlsbad, CA, USA) as described in [56]. Parallel staining with irrelevant primary antibody or secondary antibody were used to control specificity.…”

Antineoplastic drug‐loaded polymer‐modified magnetite nanoparticles: Comparative analysis of EPR‐mediated drug delivery

“…It is widely believed that ovarian carcinomas usually arise from the ovarian surface epithelium or from surface epithelial inclusion glands and cysts. Endometriosis is also considered a plausible and well‐documented, but not necessarily obligate precursor of ovarian carcinomas, particularly those with clear cell or endometrioid differentiation (Annie & Barker, 2015; Minopoli et al, 2019). Pathologists currently employ a classification system for ovarian carcinomas based entirely on light microscopic analysis of tumor cell morphology (Lisio, Fu, Goyeneche, Gao, & Telleria, 2019).…”