Periostin promotes ovarian cancer metastasis by enhancing M2 macrophages and cancer-associated fibroblasts via integrin-mediated NF-κB and TGF-β2 signaling

Abstract

Background Ovarian cancer has the highest mortality among gynecological cancers due to late diagnosis and lack of effective targeted therapy. Although the study of interplay between cancer cells with their microenvironment is emerging, how ovarian cancer triggers signaling that coordinates with immune cells to promote metastasis is still elusive. Methods Microarray and bioinformatics analysis of low and highly invasive ovarian cancer cell lines were used to reveal periostin (POSTN), a matrix protein with multifunctions in cancer, with elevated expression in the highly invasive cells. Anchorage independent assay, Western blot, RNA interference, confocal analysis and neutralizing antibody treatment were performed to analyze the effects of POSTN on tumor promotion and to explore the underlying mechanism. Chemotaxis, flow cytometry and cytokine array analyses were undertaken to analyze the involvement of POSTN in cancer-associated fibroblast (CAF) and macrophage modulation. Correlations between POSTN expression levels and clinical characteristics were analyzed using the Oncomine, commercial ovarian cancer cDNA and China Medical University Hospital patient cohort. In vivo effect of POSTN on metastasis was studied using a mouse xenograft model. Results Expression of POSTN was found to be elevated in highly invasive ovarian cancer cells. We observed that POSTN was co-localized with integrin β3 and integrin β5, which was important for POSTN-mediated activation of ERK and NF-κB. Ectopic expression of POSTN enhanced whereas knockdown of POSTN decreased cancer cell migration and invasion in vitro, as well as tumor growth and metastasis in vivo. POSTN enhanced integrin/ERK/NF-κB signaling through an autocrine effect on cancer cells to produce macrophage attracting and mobilizing cytokines including MIP-1β, MCP-1, TNFα and RANTES resulting in increased chemotaxis of THP-1 monocytes and their polarization to M2 macrophages in vitro. In agreement, tumors derived from POSTN-overexpressing SKOV3 harbored more tumor-associated macrophages than the control tumors. POSTN induced TGF-β2 expression from ovarian cancer cells to promote activation of adipose-derived stromal cells to become CAF-like cells expressing alpha smooth muscle actin and fibroblast activation protein alpha. Consistently, increased CAFs were observed in POSTN overexpressing SKOV3 cells-derived metastatic tumors. In clinical relevance, we found that expression of POSTN was positively correlated with advanced-stage diseases and poor overall survival of patients. Conclusions Our study revealed a POSTN-integrin-NF-κB-mediated signaling and its involvement in enhancing M2 macrophages and CAFs, which could potentially participate in promoting tumor growth. Our results suggest that POSTN could be a useful prognosis marker and potential therapeutic target.

Single‐cell and spatial transcriptomics reveal POSTN+ cancer‐associated fibroblasts correlated with immune suppression and tumour progression in non‐small cell lung cancer

“…26 Interestingly, POSTN + CAFs (C02_POSTN) were identified at substantial amount in multiple mBrain samples, corresponding to the function of periostin in promoting tumour metastasis. 17,18,69 These results suggested the central roles of POSTN + CAFs in modulating TME to promote tumour progression, immune escape and resistance to immunotherapies, which were also described in other solid tumours. 36,44 Through spatial transcriptomic analysis, we found that expression of POSTN, COL1A1, COL1A2 and SPP1 was high in the 'Macrophage/Fibroblast' cluster, indicating close localization of POSTN + CAFs and SPP1 + macrophages.…”

“…Additionally, correlation analysis based on the scRNA‐seq data demonstrated that C02_POSTN negatively correlated with cytotoxic CD8 + T cells or NK cells, and they presented positive correlation with exhausted CD8 + T cells and SPP1 + macrophages, a cluster of M2‐like macrophages with high expression of M2 gene signatures 26 . Interestingly, POSTN + CAFs (C02_POSTN) were identified at substantial amount in multiple mBrain samples, corresponding to the function of periostin in promoting tumour metastasis 17,18,69 . These results suggested the central roles of POSTN + CAFs in modulating TME to promote tumour progression, immune escape and resistance to immunotherapies, which were also described in other solid tumours 36,44 …”

“…The association and interactions of SPP1 + macrophages with CAFs promoted the formation of desmoplastic barrier that hindered immune infiltration and limited the efficacies of ICI immunotherapies in hepatocellular carcinoma and colorectal cancer 34,70 . Previous studies reported that periostin secreted by tumour cells acted as a chemoattractant to enhance the recruitment and polarization of M2 TAMs in glioblastoma and ovarian cancer through periostin‐integrin mediated signaling 69,71 . We predicted the potential interaction of SPP1 + macrophages and POSTN + CAFs through NicheNet 60 .…”

Construction of a hypoxia-immune-related prognostic panel based on integrated single-cell and bulk RNA sequencing analyses in gastric cancer

“…In glioma, POSTN may regulate resistance to anti-VEGF-A therapy by upregulating the expression of TGFb1 and HIF1a (22). POSTN is implicated in promoting metastasis of ovarian cancer via its ability to enhance M2 macrophages and cancer-associated fibroblasts through integrin-mediated activation of the NF-kB and TGF-b2 signaling pathways (23). Zhong et al reported that BMP4 may play an important role in regulating glycolysis in hepatocellular carcinoma cells under hypoxia and hypoglycemia (24).…”

Cancer-associated fibroblasts in gynecological malignancies: are they really allies of the enemy?

“…According to the latest Global Cancer Observatory (GLOBOCAN) report in 2020, gynecological cancers were the cause of 4,429,323 deaths and an additional 9,227,484 new cases were registered in the world ( 2 ), making them a problem of global concern in public health. Cancer-associated fibroblasts (CAFs), among other cellular and molecular components of the tumor microenvironment (TME), favor these neoplasms to occupy the first places in mortality and incidence rates ( 3 , 4 ).…”