Project description:Cancer is a rapidly evolving, multifactorial disease that accumulates numerous genetic alterations. This results in phenotypic and molecular heterogeneity within the tumor, the complexity of which is further amplified through specific interactions between cancer cells. We aimed at dissecting the molecular mechanisms underlying the cooperation between different clones. We produced clonal cell lines derived from the MDA-MB-231 breast cancer cell line, using the UbC-StarTrack system, which allows tracking of multiple clones by colour: GFP C3, mKO E10 and Sapphire D7. Characterization of these clones in vitro revealed clear differences at genetic levels, that induce differences in growth rate, morphology and cytokine expression among them. In vivo, all clonal cell lines were able to form tumors, however an injection of an equal mix of clones, led to the formation of tumors where mKO E10 was almost depleted. Additionally, the mKO E10 clonal cell line showed a significant deficiency in the formation of lung metastasis. These results confirm that even in stable cell lines heterogeneity is present. In vitro the complementation of growth medium with medium or exosomes from either parental or clonal cell lines, increased the growth rate of the other clones. Co-growth and co-injection of mKO E10 and GFP C3 clonal cell lines increased the efficiency of invasion and migration, respectively. These findings support a model where the interplay of clones confers aggressiveness and may allow to identify factors involved in cellular communication which can be involved in clonal cooperation and be new targets preventing tumor progression.
Project description:Human adipose mesenchymal stem cells are a heterogeneous population, where cell cultures derived from single cell-expanded clones present varying degrees of differential plasticity. This work focuses on the immunomodulatory/anti-inflammatory properties of these cells. To this end, 5 single cell clones were isolated (1.X and 3.X) from 2 volunteers. Regarding the expression level of the lineage-characteristic surface antigens, clones 1.10 and 1.22 expressed the lowest amounts, while clones 3.10 and 3.5 expressed more CD105 than the rest and clone 1.7 expressed higher amounts of CD73 and CD44. Regarding cytokine secretion, all clones were capable of spontaneously releasing high levels of IL-6 and low to moderate levels of IL-8. Furthermore, clone 3.X produced the highest amounts of pro-inflammatory cytokines such as IL-1β, while clones 1.10 and 1.22 highly expressed IL-4 and IL-5. These differences can be explained in part by the distinct DNA promoter methylation profile exhibited by the clones. The results of this work indicates that this stem cell population is heterogeneous in its secretion profile, and that isolation, characterization and selection of the appropriate cell clone is a more exact method for the possible treatment of different patients or pathologies.
Project description:Human adipose mesenchymal stem cells are a heterogeneous population, where cell cultures derived from single cell-expanded clones present varying degrees of differential plasticity. This work focuses on the immunomodulatory/anti-inflammatory properties of these cells. To this end, 5 single cell clones were isolated (1.X and 3.X) from 2 volunteers. Regarding the expression level of the lineage-characteristic surface antigens, clones 1.10 and 1.22 expressed the lowest amounts, while clones 3.10 and 3.5 expressed more CD105 than the rest and clone 1.7 expressed higher amounts of CD73 and CD44. Regarding cytokine secretion, all clones were capable of spontaneously releasing high levels of IL-6 and low to moderate levels of IL-8. Furthermore, clone 3.X produced the highest amounts of pro-inflammatory cytokines such as IL-1β, while clones 1.10 and 1.22 highly expressed IL-4 and IL-5. These differences can be explained in part by the distinct DNA promoter methylation profile exhibited by the clones. The results of this work indicates that this stem cell population is heterogeneous in its secretion profile, and that isolation, characterization and selection of the appropriate cell clone is a more exact method for the possible treatment of different patients or pathologies. Total DNA isolated by standard procedures from different clones of human dipose mesenchymal stem cells (hMSC)
Project description:Transcriptional profiling comparing tumoral and non tumoral part of human liver infected by hepatitis B virus Expression profiling of induced hepatitis B virus tumor from human liver and expression profiling of hepatitis B virus infected non tumoral part of the liver from french patients Please note that there are 62 raw files (i.e.'AFE_raw_files.tar' linked to the Series records) for 124 samples since two samples were on the same array, one in Cy3 and one in Cy5. The corresponding raw data file for each sample is indicated in the Sample description field. This dataset is part of the TransQST collection.
Project description:To study the interaction between mutated clones using interspecies competition and mathematical models Methods: RNA was extracted from 3-D cultures after 48 hours of growth in spheroids using manufacturer protocol (Qiagen) and sequenced in duplicate using Illumina NextSeq500. libraries were prepared from 500ng of purified RNA using Illumina TruSeq preparation kits. Sequence reads that passed quality filters were aligned to the UCSC mm9 reference genome and quanitifed using STAR (v2.5.1b). Normalized read counts (FPKM) were calculated using cufflinks and transcriptome profiling was analyzed using DESeq2. We determined that previously sensitive cancer cell clones within heterogeneous tumors will induce drug resistant phenotypes among neighboring cells while under drug pressure Non-mutational drug resistance can arise through drug-induced mechanisms augmented in heterogeneous tumors.
Project description:We used vertical growth phase (VGP) human VMM5 melanoma cells to ask whether the tumor microenvironment could induce matrix metalloproteinase-1 (MMP-1) in vivo, and whether this induction correlated with metastasis. We isolated two clones from parental VMM5 cells: a low MMP-1 producing clone (C4) and high producing clone (C9). When these clones were injected orthotopically (intradermally) into nude mice, both were equally tumorigenic and produced equivalent and abundant amounts of MMP-1. However, the tumors from the C4 clones displayed different growth kinetics and distinct profiles of gene expression from the C9 population. The C4 tumors, which had low MMP-1 levels in vitro, appeared to rely on growth factors and cytokines in the microenvironment to increase MMP-1 expression in vivo, while MMP-1 levels remained constant in the C9 tumors. C9 cells, but not C4 cells, grew as spheres in culture and expressed higher levels of JARID 1B, a marker associated with melanoma initiating cells. We conclude that VMM5 melanoma cells exhibit striking intra-tumor heterogeneity, and that the tumorigenicity of these clones is driven by different molecular pathways. Our data suggest that there are multiple mechanisms for melanoma progression within a tumor, which may require different therapeutic strategies.
Project description:Transcriptional profiling of pancreatic tumors comparing to adjacent non-tumoral pancreatic tissue by two strategies. The first strategy implies a traditional microarray assay, while the second involves a previous Suppression Subtractive Hybridization (SSH) and then the traditional microarray assay. Goal was to determine differences in gene expression profile in tumor samples by both strategies. Two-condition experiment, tumoral vs. non-tumoral tissues. Biological replicates: 6 tumoral replicates, 6 non-tumoral replicates. The same replicates for both strategies.