A new murine model of osteoblastic/osteolytic lesions from human androgen-resistant prostate cancer

BACKGROUND: Up to 80% of patients dying from prostate carcinoma have developed bone metastases that are incurable. Castration is commonly used to treat prostate cancer. Although the disease initially responds to androgen blockade strategies, it often becomes castration-resistant (CRPC for Castration Resistant Prostate Cancer). Most of the murine models of mixed lesions derived from prostate cancer cells are androgen sensitive. Thus, we established a new model of CRPC (androgen receptor (AR) negative) that causes mixed lesions in bone. METHODS: PC3 and its derived new cell clone PC3c cells were directly injected into the tibiae of SCID male mice. Tumor growth was analyzed by radiography and histology. Direct effects of conditioned medium of both cell lines were tested on osteoclasts, osteoblasts and osteocytes. RESULTS: We found that PC3c cells induced mixed lesions 10 weeks after intratibial injection. In vitro, PC3c conditioned medium was able to stimulate tartrate resistant acid phosphatase (TRAP)-positive osteoclasts. Osteoprotegerin (OPG) and endothelin-1 (ET1) were highly expressed by PC3c while dikkopf-1 (DKK1) expression was decreased. Finally, PC3c highly expressed bone associated markers osteopontin (OPN), Runx2, alkaline phosphatase (ALP), bone sialoprotein (BSP) and produced mineralized matrix in vitro in osteogenic conditions. CONCLUSIONS: We have established a new CRPC cell line as a useful system for modeling human metastatic prostate cancer which presents the mixed phenotype of bone metastases that is commonly observed in prostate cancer patients with advanced disease. This model will help to understand androgen-independent mechanisms involved in the progression of prostate cancer in bone and provides a preclinical model for testing the effects of new treatments for bone metastases

Study of the expression of a Small Leucin-Rich Proteoglycan, Asporin, in normal human osteoblasts and regulation by breast cancer cells

Dottorato di Ricerca in Biochimica Cellulare ed attività dei Farmaci in Oncologia, Ciclo XXVII, a.a. 2014Asporin (ASPN) is an extracellular matrix protein that belongs to the Small Leucine Rich Repeat proteoglycan (SLRP) family. Asporin is abundantly expressed in the articular cartilage of individuals with osteoarthritis. In the context of osteoarthritis, several studies have shown that asporin regulates cartilage matrix gene expression and cartilage formation by modulating the transforming growth factor-β (TGF- β) signaling pathway. Asporin directly binds to TGF‐β and inhibits TGF-β-mediated expression of cartilage matrix genes. Previous studies in our laboratory, showed that Asporin inhibits TGF- β-1-mediated SMAD2 phosphorylation in breast cancer cells as well as migration and epithelial to mesenchymal transition in A549 human lung cancer cells. The present study was undertaken to investigate whether asporin secretion could indirectly mediate the ability of metastatic breast cancer cells to regulate osteoblastic differentiation. The Wnt antagonist sclerostin (SOST) is a potent inhibitor of bone formation. We considered the possibility that the balance between ASPN and SOST present in the ECM may create a specific environment favorable to aggressive breast cancer cell growth. Results: Breast cancer cells do not produce ASPN themselves but they regulate its expression in osteoblasts. Normal human osteoblasts have been cultured in presence of MCF7 and MDA-MB-231 serum-free conditioned medium. Immunoblot analysis and real time PCR, revealed a significant increase in ASPN expression and secretion in osteoblasts treated with MCF7-conditioned medium, while the opposite effect was observed with MDA-MB-231-conditioned medium. We investigated the role of MCF7 and MDAMB231 conditioned media in osteoblast differentiation and mineralization through alkaline phospatase and Runx2 expression. Our results showed the ability of MCF7 conditioned medium to induce the osteoblast differentiation and mineralization compared to the MDA-MB-231 conditioned medium treatment. Osteoblasts treated with MCF7 conditioned medium and challenged with recombinant SOST showed a significant reduction in their differentiation potential through the decrease of ASPN expression. Contrarily to non-metastatic MCF-7 breast cancer cells, MDA-MB-231 metastatic breast cancer cells inhibited the secretion of ASPN by osteoblasts through the overexpression of SOST. The result is the reduction of osteoblast differentiation and mineralization that can create a specific environment favorable to aggressive breast cancer cell growthUniversità degli Studi della Calabri

L’asporine : une nouvelle défense naturelle contre le cancer du sein

International audienc

Cancer-Associated Fibroblast Diversity Shapes Tumor Metabolism in Pancreatic CancerCancer-Associated Fibroblast Diversity Shapes Tumor Metabolism in Pancreatic Cancer

Despite extensive research, the 5-year survival rate of pancreatic cancer (PDAC) patients remains at only 9%. Patients often show poor treatment response, due partly to a highly complex tumor microenvironment (TME). Cancer-associated fibroblast (CAF) heterogeneity is characteristic of the pancreatic TME, where several CAF subpopulations have been identified, such as myofibroblastic CAFs (myCAFs), inflammatory CAFs (iCAFs), and antigen presenting CAFs (apCAFs). In PDAC, cancer cells continuously adapt their metabolism (metabolic switch) to environmental changes in pH, oxygenation, and nutrient availability. Recent advances show that these environmental alterations are all heavily driven by stromal CAFs. CAFs and cancer cells exchange cytokines and metabolites, engaging in a tight bidirectional crosstalk, which promotes tumor aggressiveness and allows constant adaptation to external stress, such as chemotherapy. In this review, we summarize CAF diversity and CAF-mediated metabolic rewiring, in a PDAC-specific context. First, we recapitulate the most recently identified CAF subtypes, focusing on the cell of origin, activation mechanism, species-dependent markers, and functions. Next, we describe in detail the metabolic crosstalk between CAFs and tumor cells. Additionally, we elucidate how CAF-driven paracrine signaling, desmoplasia, and acidosis orchestrate cancer cell metabolism. Finally, we highlight how the CAF/cancer cell crosstalk could pave the way for new therapeutic strategies

Cancer-Associated Fibroblast Diversity Shapes Tumor Metabolism in Pancreatic Cancer

Despite extensive research, the 5-year survival rate of pancreatic cancer (PDAC) patients remains at only 9%. Patients often show poor treatment response, due partly to a highly complex tumor microenvironment (TME). Cancer-associated fibroblast (CAF) heterogeneity is characteristic of the pancreatic TME, where several CAF subpopulations have been identified, such as myofibroblastic CAFs (myCAFs), inflammatory CAFs (iCAFs), and antigen presenting CAFs (apCAFs). In PDAC, cancer cells continuously adapt their metabolism (metabolic switch) to environmental changes in pH, oxygenation, and nutrient availability. Recent advances show that these environmental alterations are all heavily driven by stromal CAFs. CAFs and cancer cells exchange cytokines and metabolites, engaging in a tight bidirectional crosstalk, which promotes tumor aggressiveness and allows constant adaptation to external stress, such as chemotherapy. In this review, we summarize CAF diversity and CAF-mediated metabolic rewiring, in a PDAC-specific context. First, we recapitulate the most recently identified CAF subtypes, focusing on the cell of origin, activation mechanism, species-dependent markers, and functions. Next, we describe in detail the metabolic crosstalk between CAFs and tumor cells. Additionally, we elucidate how CAF-driven paracrine signaling, desmoplasia, and acidosis orchestrate cancer cell metabolism. Finally, we highlight how the CAF/cancer cell crosstalk could pave the way for new therapeutic strategies

Anticancer, antiplasmodial and antitrypanosomal activities of crude extracts of Platanus Orientalis

The present study evaluates the anticancer, antiplasmodial and antitrypanosomal potential of leaf extract of the buds and bark of Platanus orientalis. The buds and bark of the plant were extracted with dichloromethane. Concerning the cytotoxic activity, the two extracts of the plant have shown a good growth inhibition of cervical adenocarcinoma cell lines (HeLa cells), with an IC-50 of 10.1 μg/mL and 30.9 μg/mL, respectively. The activity was slightly lower for breast adenocarcinoma cell lines (MCF-7), with an IC-50 of 38.6 μg/mL and 31.6 μg/mL, respectively. The IC50 values were superior to 100 µg/mL for MDAMB-231 cells. The extracts showed also antiplasmodial (12.7 μg/mL for bark extracts and 35.3 μg/mL for buds extracts) and antitrypnosomal activities (20.0 μg/mL for bark extracts and 21.8 μg/mL for buds extracts)

Cell Membrane Proteomic Analysis Identifies Proteins Differentially Expressed in Osteotropic Human Breast Cancer Cells12

Metastatic breast cancer cells are characterized by their high propensity to colonize the skeleton and form bone metastases, causing major morbidity and mortality. Identifying key proteins involved in the osteotropic phenotype would represent a major step toward the development of both new prognostic markers and new effective therapies. Cell surface proteins differentially expressed in cancer cells are preferred potential targets for antibody-based targeted therapies. In this study, using cell surface biotinylation and a mass spectrometric approach, we have compared the profile of accessible cell surface proteins between the human breast cancer cell line MDA-MB-231 and its highly osteotropic B02 subclone. This strategy allowed the identification of several proteins either up- or downregulated in the osteotropic cell line, and differential protein expressions were validated using antibody-based techniques. Class I HLAs were down-regulated in the bone metastatic variant, whereas αvβ3 integrins, among others, were consistently up-regulated in this latter cell line. These results show that comprehensive profiling of the cell surface proteome of mother cancerous cell lines and derived organ-specific metastatic cell lines provides an effective approach for the identification of potential accessible marker proteins for both prognosis and antibody-based targeted therapies

Dual function of ERR alpha in breast cancer and bone metastasis formation: implication of VEGF and osteoprotegerin.

Bone metastasis is a complication occurring in up to 70% of advanced breast cancer patients. The estrogen receptor-related receptor alpha (ERRalpha) has been implicated in breast cancer and bone development, prompting us to examine whether ERRalpha may function in promoting the osteolytic growth of breast cancer cells in bone. In a mouse xenograft model of metastatic human breast cancer, overexpression of wild-type ERRalpha reduced metastasis, whereas overexpression of a dominant negative mutant promoted metastasis. Osteoclasts were directly affected and ERRalpha upregulated the osteoclastogenesis inhibitor, osteoprotegerin (OPG), providing a direct mechanistic basis for understanding how ERRalpha reduced breast cancer cell growth in bone. In contrast, ERRalpha overexpression increased breast cancer cell growth in the mammary gland. ERRalpha-overexpressing primary tumors were highly vascularized, consistent with an observed upregulation of angiogenic growth factor, the VEGF. In support of these findings, we documented that elevated expression of ERRalpha mRNA in breast carcinomas was associated with high expression of OPG and VEGF and with disease progression. In conclusion, our results show that ERRalpha plays a dual role in breast cancer progression in promoting the local growth of tumor cells, but decreasing metastatic growth of osteolytic lesions in bone

Myoferlin is a key regulator of EGFR activity in breast cancer.

Myoferlin is a member of the ferlin family of proteins that participate in plasma membrane fusion, repair and endocytosis. While some reports have implicated myoferlin in cancer, the extent of its expression in and contributions to cancer are not well established. In this study, we show that myoferlin is overexpressed in human breast cancers and that it is has a critical role in controlling degradation of the EGFR after its activation and internalization in breast cancer cells. Myoferlin depletion blocked EGF-induced cell migration and epithelial-to-mesenchymal transition. Both effects were induced as a result of impaired degradation of phosphorylated EGFR via dysfunctional plasma membrane caveolae and alteration of caveolin homooligomerization. In parallel, myoferlin depletion reduced tumor development in a chicken chorioallantoic membrane xenograft model of human breast cancer. Considering the therapeutic significance of EGFR targeting, our findings identify myoferlin as an novel candidate function to target for future drug development

Paladin, overexpressed in colon cancer, is required for actin polymerisation and liver metastasis dissemination

Introduction: Colorectal cancer remains a public health issue and most colon cancer patients succumb to the development of metastases. Using a specific protocol of pressure-assisted interstitial fluid extrusion to recover soluble biomarkers, we identified paladin as a potential colon cancer liver metastases biomarker. Methods: Using shRNA gene knockdown, we explored the biological function of paladin in colon cancer cells and investigated the phospho-proteome within colon cancer cells. We successively applied in vitro migration assays, in vivo metastasis models and co-immunoprecipitation experiments. Results: We discovered that paladin is required for colon cancer cell migration and metastasis, and that paladin depletion altered the phospho-proteome within colon cancer cells. Data are available via ProteomeXchange with identifier PXD030803. Thanks to immunoprecipitation experiments, we demonstrated that paladin, was interacting with SSH1, a phosphatase involved in colon cancer metastasis. Finally, we showed that paladin depletion in cancer cells results in a less dynamic actin cytoskeleton. Conclusions: Paladin is an undervalued protein in oncology. This study highlights for the first time that, paladin is participating in actin cytoskeleton remodelling and is required for efficient cancer cell migration