0000000001315205
AUTHOR
Chiara Ratti
Additional file 1 of Intracellular osteopontin protects from autoimmunity-driven lymphoma development inhibiting TLR9-MYD88-STAT3 signaling
Additional file 1. Supplementary file 1. Supplementary Material & methods.
Additional file 6 of Intracellular osteopontin protects from autoimmunity-driven lymphoma development inhibiting TLR9-MYD88-STAT3 signaling
Additional file 6: Supplementary Figure S3. Immunohistochemistry staining of OPN IHC for OPN was performed in Fas lpr/lpr and OPN-/-Fas lpr/lpr mice with either no lymphoma or with lymphomatous cells. As expected, no staining is detected in case of OPN-deficient mice.
Intracellular osteopontin protects from autoimmunity-driven lymphoma development inhibiting TLR9-MYD88-STAT3 signaling
Abstract Background Autoimmune disorders, including Systemic Lupus Erythematosus (SLE), are associated with increased incidence of hematological malignancies. The matricellular protein osteopontin (OPN) has been linked to SLE pathogenesis, as SLE patients show increased serum levels of OPN and often polymorphisms in its gene. Although widely studied for its pro-tumorigenic role in different solid tumours, the role of OPN in autoimmunity-driven lymphomagenesis has not been investigated yet. Methods To test the role of OPN in the SLE-associated lymphomagenesis, the SLE-like prone Faslpr/lpr mutation was transferred onto an OPN-deficient background. Spleen from Faslpr/lpr and OPN-/-Faslpr/lpr …
Additional file 4 of Intracellular osteopontin protects from autoimmunity-driven lymphoma development inhibiting TLR9-MYD88-STAT3 signaling
Additional file 4: Supplementary Figure S1. Evaluation of autoimmunity in Faslpr/lpr and OPN-/-Faslpr/lpr mice. A. Quantification of OPN in sera from Faslpr/lpr mice at 2 (n=8) and 5 months of age (n=7) by ELISA. Sera from BALB/c and OPN-/- mice were tested as controls. Data are expressed as ng/ml and are a pool of 2 experiments (*, P<0.05; Ordinary one way ANOVA). B. Flow cytometry analysis showing the relative number of splenic autoimmune CD3+B220+ T cells in Faslpr/lpr (n=15) and OPN-/-Faslpr/lpr mice (n=18) and at about 5-6 months of age. The graph shows a pool of 3 different experiments (***, P<0.001; Student t test). C. Representative spleen photograph from BALB/c, OPN-/-, Faslp…
Additional file 7 of Intracellular osteopontin protects from autoimmunity-driven lymphoma development inhibiting TLR9-MYD88-STAT3 signaling
Additional file 7: Supplementary Figure S4. Characterization of OPL239 and OPL241 DLBCL cell lines. A. Flow cytometry analysis showing the expression of B220, IgM, IgD and IgA in OPL239 and OPL241 cell lines. B. Hardy’s multiparametric flow cytometry panel illustrating the expression of CD93, CD21/35 and CD23 on OPL239 and OPL241 cell lines. C. Flow cytometry analysis showing the expression of TLR9 on OPL239 and OPL241 cell lines. D. RT-PCR analysis showing Spp1 mRNA level in overexpressing cell variants. E. Western blot for OPN protein expression (in presence or not of BFA, that blocks protein secretion) in parental and IRES-Green-based cell variants. 4T1 mammary cell line was used as posi…
Osteopontin shapes immunosuppression in the metastatic niche.
Abstract The matricellular protein osteopontin (OPN, Spp-1) is widely associated with cancer aggressiveness when produced by tumor cells, but its impact is uncertain when produced by leukocytes in the context of the tumor stroma. In a broad study using Spp1−/− mice along with gene silencing in tumor cells, we obtained evidence of distinct and common activities of OPN when produced by tumor or host cells in a spontaneously metastatic model of breast cancer. Different cellular localization of OPN is associated with its distinct activities, being mainly secreted in tumor cells while intracellular in myeloid cells. OPN produced by tumor cells supported their survival in the blood stream, wherea…
Trabectedin Overrides Osteosarcoma Differentiative Block and Reprograms the Tumor Immune Environment Enabling Effective Combination with Immune Checkpoint Inhibitors.
Abstract Purpose: Osteosarcoma, the most common primary bone tumor, is characterized by an aggressive behavior with high tendency to develop lung metastases as well as by multiple genetic aberrations that have hindered the development of targeted therapies. New therapeutic approaches are urgently needed; however, novel combinations with immunotherapies and checkpoint inhibitors require suitable preclinical models with intact immune systems to be properly tested. Experimental Design: We have developed immunocompetent osteosarcoma models that grow orthotopically in the bone and spontaneously metastasize to the lungs, mimicking human osteosarcoma. These models have been used to test the effica…
Additional file 5 of Intracellular osteopontin protects from autoimmunity-driven lymphoma development inhibiting TLR9-MYD88-STAT3 signaling
Additional file 5: Supplementary Figure S2. Evaluation of the different spenic B cell subsets. A. Example of Hardy’s gating strategy to discern the different CD93+ immature (Transitional T1, T2, T3) and CD93- mature [follicular B (FOB), marginal zone B (MZB) and CD21/35-CD23-] B cell subsets in the spleen from a BALB/c mouse. B. Flow cytometry analysis based on Hardy’s multiparametric panel illustrating the fraction of splenic CD23+ FOB, CD21/35+ MZB cells, and CD23-CD21/35- cells from the spleens of naive and autoimmune mice. 3 mice per group were used for the experiment. Data are referred to one representative experiment out of 3 (***, P<0.001; Two-way ANOVA) (****, P<0.0001; Two-wa…
Oncogene-driven intrinsic inflammation induces leukocyte production of tumor necrosis factor that critically contributes to mammary carcinogenesis.
Abstract Oncogene activation promotes an intrinsic inflammatory pathway that is crucial for cancer development. Here, we have investigated the actual effect of the inflammatory cytokine tumor necrosis factor (TNF) on the natural history of spontaneous mammary cancer in the HER2/neuT (NeuT) transgenic mouse model. Bone marrow transplantation from TNF knockout mice into NeuT recipients significantly impaired tumor growth, indicating that the source of TNF fostering tumor development was of bone marrow origin. We show that the absence of leukocyte-derived TNF disarranged the tumor vasculature, which lacked pericyte coverage and structural integrity, leading to diffuse vascular hemorrhage and s…
Additional file 8 of Intracellular osteopontin protects from autoimmunity-driven lymphoma development inhibiting TLR9-MYD88-STAT3 signaling
Additional file 8: Supplementary Figure S5. Expression of OPN in human GCB- and ABC-DLBCL samples. Immunohistochemistry analysis for OPN was performed on six cases for GCB- and ABC-DLBCLs. Representative images for two cases for each subtype are shown (quantification is shown in Figure 7B). Magnification 20X.
Additional file 2 of Intracellular osteopontin protects from autoimmunity-driven lymphoma development inhibiting TLR9-MYD88-STAT3 signaling
Additional file 2: Supplementary Table S1. Differentially expressed genes between CD19+ cellsfrom OPN-/-Fas lpr/lpr and Faslpr/lpr mice.
Additional file 3 of Intracellular osteopontin protects from autoimmunity-driven lymphoma development inhibiting TLR9-MYD88-STAT3 signaling
Additional file 3: Supplementary Table S2. Differentially expressed genes between CD19+ cellsfrom OPN-/-and OPN+/+ mice.