Expression profiling of secreted and cell surface genes of 5 transformed cell lines and derivative xenograft tumors
Ontology highlight
ABSTRACT: Since the early stages of tumorigenesis involve adhesion, escape from immune surveillance, vascularization and angiogenesis, we devised a strategy to study the expression profiles of all publicly known and putative secreted and cell surface genes. We designed a custom oligonucleotide microarray containing probes for 3531 secreted and cell surface genes to study 5 diverse human transformed cell lines and their derivative xenograft tumors. The origins of these human cell lines were lung (A549), breast (MDA MB-231), colon (HCT-116), ovarian (SK-OV- 3) and prostate (PC3) carcinomas. Three different analyses were performed: (1) A PCA-based linear discriminant analysis identified a 54 gene profile characteristic of all tumors when pooled tumor data were analyzed, (2) Application of MANOVA (P Pcorr corr < .05) to pooled tumor data revealed a larger set of 149 differentially expressed genes. (3) After MANOVA was performed on data from individual tumors, a comparison of differential genes amongst all tumor types revealed 12 common differential genes. Seven of the 12 genes were identified by all three analytical methods. These included late angiogenic, morphogenic and extracellular matrix genes such as ANGPTL4, COL1A1, GP2, GPR57, LAMB3, PCDHB9 and PTGER3. The differential expression of ANGPTL4 and COL1A1 and other genes was confirmed by quantitative PCR. Overall, a comparison of the three analyses revealed an expression pattern indicative of late angiogenic processes. These results show that a xenograft model using multiple cell lines of diverse tissue origin can identify common tumorigenic cell surface or secreted molecules that may be important biomarker and therapeutic discoveries.
Project description:Since the early stages of tumorigenesis involve adhesion, escape from immune surveillance, vascularization and angiogenesis, we devised a strategy to study the expression profiles of all publicly known and putative secreted and cell surface genes. We designed a custom oligonucleotide microarray containing probes for 3531 secreted and cell surface genes to study 5 diverse human transformed cell lines and their derivative xenograft tumors. The origins of these human cell lines were lung (A549), breast (MDA MB-231), colon (HCT-116), ovarian (SK-OV- 3) and prostate (PC3) carcinomas. Three different analyses were performed: (1) A PCA-based linear discriminant analysis identified a 54 gene profile characteristic of all tumors when pooled tumor data were analyzed, (2) Application of MANOVA (P Pcorr corr < .05) to pooled tumor data revealed a larger set of 149 differentially expressed genes. (3) After MANOVA was performed on data from individual tumors, a comparison of differential genes amongst all tumor types revealed 12 common differential genes. Seven of the 12 genes were identified by all three analytical methods. These included late angiogenic, morphogenic and extracellular matrix genes such as ANGPTL4, COL1A1, GP2, GPR57, LAMB3, PCDHB9 and PTGER3. The differential expression of ANGPTL4 and COL1A1 and other genes was confirmed by quantitative PCR. Overall, a comparison of the three analyses revealed an expression pattern indicative of late angiogenic processes. These results show that a xenograft model using multiple cell lines of diverse tissue origin can identify common tumorigenic cell surface or secreted molecules that may be important biomarker and therapeutic discoveries. Keywords: parallel sample
Project description:Angiopoietin-like 4 (ANGPTL4) is known to regulate various cellular and systemic functions. However, its cell-specific role in endothelial cells (ECs) function and metabolic homeostasis remains to be elucidated. Here, using endothelial-specific Angptl4 knock-out mice (Angptl4iEC), and transcriptomics and metabolic flux analysis, we demonstrate that ANGPTL4 is required for maintaining EC metabolic function vital for vascular permeability and angiogenesis. Knockdown of ANGPTL4 in ECs promotes lipase-mediated lipoprotein lipolysis, which results in increased fatty acid (FA) uptake and oxidation. This is also paralleled by a decrease in proper glucose utilization for angiogenic activation of ECs. Mice with endothelial-specific deletion of Angptl4 showed decreased pathological neovascularization with stable vessel structures characterized by increased pericyte coverage and reduced permeability. Together, our study denotes the role of endothelial-ANGPTL4 in regulating cellular metabolism and angiogenic functions of EC.
Project description:Malignant glioblastoma (GBM) is a highly aggressive brain tumor with a dismal prognosis and limited therapeutic options. Genomic profiling of GBM samples in the TCGA database has identified four molecular subtypes (Proneural, Neural, Classical and Mesenchymal), which may arise from different glioblastoma stem-like cell (GSC) populations. In the present study, we identify two GSC populations that produce GBM tumors by subcutaneous and intracranial injection with identical histological features. Gene expression analysis revealed that xenografts of GSCs grown as spheroid cultures had a Classical molecular subtype similar to that of bulk tumor cells. In contrast xenografts of GSCs grown as adherent cultures on laminin-coated plates expressed a Mesenchymal gene signature. Adherent GSC-derived xenografts had high STAT3 and ANGPTL4 expression as well as enrichment for stem cell markers, transcriptional networks and pro-angiogenic markers characteristic of the Mesenchymal subtype. Examination of clinical samples from GBM patients showed that STAT3 expression was directly correlated with ANGPTL4 expression, and that increased expression of these genes correlated with poor patient survival and performance. A pharmacological STAT3 inhibitor abrogated STAT3 binding to the ANGPTL4 promoter and exhibited anticancer activity in vivo. Taken together, we identified two distinct GSC populations that produce histologically identical tumors but with very different gene expression patterns, and a STAT3/ ANGPTL4 pathway in glioblastoma that may serve as a target for therapeutic intervention. 2 samples of each variable were analyzed. Cells were cultured under normal adherent conditon (Bulk tumor cells), non-adherent plates with stem cell medium (Sp-GSC) or laminin-coated plates with stem cell medium (Ad-GSC). Xenografts were generated in NSG mice by subcutaneous inoculation.
Project description:Malignant glioblastoma (GBM) is a highly aggressive brain tumor with a dismal prognosis and limited therapeutic options. Genomic profiling of GBM samples in the TCGA database has identified four molecular subtypes (Proneural, Neural, Classical and Mesenchymal), which may arise from different glioblastoma stem-like cell (GSC) populations. In the present study, we identify two GSC populations that produce GBM tumors by subcutaneous and intracranial injection with identical histological features. Gene expression analysis revealed that xenografts of GSCs grown as spheroid cultures had a Classical molecular subtype similar to that of bulk tumor cells. In contrast xenografts of GSCs grown as adherent cultures on laminin-coated plates expressed a Mesenchymal gene signature. Adherent GSC-derived xenografts had high STAT3 and ANGPTL4 expression as well as enrichment for stem cell markers, transcriptional networks and pro-angiogenic markers characteristic of the Mesenchymal subtype. Examination of clinical samples from GBM patients showed that STAT3 expression was directly correlated with ANGPTL4 expression, and that increased expression of these genes correlated with poor patient survival and performance. A pharmacological STAT3 inhibitor abrogated STAT3 binding to the ANGPTL4 promoter and exhibited anticancer activity in vivo. Taken together, we identified two distinct GSC populations that produce histologically identical tumors but with very different gene expression patterns, and a STAT3/ ANGPTL4 pathway in glioblastoma that may serve as a target for therapeutic intervention.
Project description:Webb2002 - Fas/FasL mediated tumor T-cell interaction
This deterministic model of
immunological surveillance involving tumour cell–T-lymphocyte
interaction, cell surface expression of Fas/FasL, and their
secreted soluble forms.
This model is described in the article:
Cells behaving badly: a
theoretical model for the Fas/FasL system in tumour
immunology.
Webb SD, Sherratt JA, Fish RG.
Math Biosci 2002 Sep-Oct; 179(2):
113-129
Abstract:
One proposed mechanism of tumour escape from immune
surveillance is tumour up-regulation of the cell surface ligand
FasL, which can lead to apoptosis of Fas receptor (Fas)
positive lymphocytes. Based upon this 'counterattack', we have
developed a mathematical model involving tumour cell-lymphocyte
interaction, cell surface expression of Fas/FasL, and their
secreted soluble forms. The model predicts that (a) the
production of soluble forms of Fas and FasL will lead to the
down-regulation of the immune response; (b) matrix
metalloproteinase (MMP) inactivation should lead to increased
membrane FasL and result in a higher rate of Fas-mediated
apoptosis for lymphocytes than for tumour cells. Recent studies
on cancer patients lend support for these predictions. The
clinical implications are two-fold. Firstly, the use of broad
spectrum MMP inhibitors as anti-angiogenic agents may be
compromised by their adverse effect on tumour FasL
up-regulation. Also, Fas/FasL interactions may have an impact
on the outcome of numerous ongoing immunotherapeutic trials
since the final common pathway of all these approaches is the
transduction of death signals within the tumour cell.
This model is hosted on
BioModels Database
and identified by:
BIOMD0000000661.
To cite BioModels Database, please use:
Chelliah V et al. BioModels: ten-year
anniversary. Nucl. Acids Res. 2015, 43(Database
issue):D542-8.
To the extent possible under law, all copyright and related or
neighbouring rights to this encoded model have been dedicated to
the public domain worldwide. Please refer to
CC0
Public Domain Dedication for more information.
Project description:Renal cell carcinoma (RCC), the most common form of kidney cancer, is a heterogeneous disease with clear cell RCC (ccRCC) being the most prevalent and aggressive subtype. While most ccRCC tumors have elevated expression of angiopoietin-like 4 (ANGPTL4), in our study we identified a significant subset of patients whose cancers show no increase in ANGPTL4 expression. These patients have a worse prognosis compared to the patients with high expression of ANGPTL4. These ANGPTL4-low cancers are characterized by the increased frequency of wild-type Von Hippel-Lindau (wt VHL), a gene that is commonly mutated in ccRCC, and an enrichment for genes associated with lipid metabolism. Using RCC tumor models with wild type VHL, we demonstrate that ANGPTL4 behaves as a tumor suppressor. The loss of ANGPTL4 in ccRCC cell lines results in increased tumor growth and colony formation in a lysosomal acid lipase (LAL)-dependent manner, a phenotype rescued by the expression of N-terminus ANGPTL4. At the mechanistic level, the loss of ANGPTL4 increases lysosomal acid lipase activity in ccRCC cells. This data suggests that ANGPTL4 enacts its tumor-suppressive effects in ccRCC by regulating LAL activity. Importantly, the identified patient cohort with low ANGPTL4 expression may exhibit increased reliance on lipid metabolism, which can be a point of target for future therapy.
Project description:Interventions: Gold Standard:tissue or cytopathology;Index test:secreted LncRNA
Primary outcome(s): Differential expression of LncRNA in peripheral blood exosomes of patients
Study Design: Single arm
Project description:Background: Gastric cancer (GC) is a serious malignant disease of digestive system over the world. Angiopoietin-like protein 4 (ANGPTL4) was a secreted glycoprotein, which involved in multiple pathophysiological process, such as metabolic reprogramming, angiogenesis, proliferation and metastasis. Nevertheless, the expression, function, and prognostic significance of ANGPTL4 in GC are still contradictory. Methods: Bioinformatic analysis to confirm the expression and prognosis of ANGPTLs in GC. IF staining was used to detect ANGPTL4 protein in GC samples. The molecular biological effect of ANGPTL4 was confirmed by EdU, MTT, FCS, wound healing, transwell, tube formation chorioallantoic membrane model, and nude mice model assay in GC cell lines. RNA-seq was used to confirm the potential downstream mechanism of ANGPTL4, which verified by PCR, WB, IF, co-IP and molecular docking. Results: ANGPTL4, as an important ANGPTLs, was significantly correlated with poor diagnosis in GC patients. The proliferation, migration, invasion, apoptosis escaping, angiogenesis ability was enhanced by ANGPTL4 in SNU5 and MKN7, but repressed in AGS. LGALS7 could be upregulated in AGS transfected with ANGPTL4 OE, which was one of the key factors that cause ANGPTL4 to produce an anticancer phenotype. Conclusion: LGALS7 was a key factor for the contradictory effect of ANGPTL4 on activating hedgehog pathway to driving GC progression, including proliferation, migration, apoptosis escaping, angiogenesis and lymphangiogenesis.