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: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.
Project description:Purpose: The most clearly established genetic hallmark in oligodendroglial tumors (OTs) is the combined loss of 1p/19q, a molecular alteration characteristic of tumors responding to therapy. Markers of tumoral progression have not yet been studied. Experimental Design: Novel markers of tumoral progression in OTs were sought through gene expression profiling analysis. Results: Unsupervised hierarchical cluster analysis classified OTs into two main groups associated with tumoral grade, independent of histological subtype and 1p/19q status. Differential gene expression analysis between low- and high-grade OTs revealed that only cell cycle-related genes were significantly upregulated in high-grade OTs. Among the deregulated genes, NDRG2 downregulation was detected in high-grade OTs with combined loss of 1p/19q. Expression analysis revealed low transcript levels of NDRG2 relative to non-tumoral brain tissue in 45% (9/20) of high-grade OTs. Furthermore, the low transcript levels of NDRG2 were significantly associated with a worse clinical outcome in patients. Transcript levels of NDRG2 were associated with promoter hypermethylation, which was detected in 38.4% (10/26) of high-grade OTs. The treatment of glioma cell lines T98 and LN18 with demethylating agents increased the mRNA expression levels of NDRG2 relative to the control cell line. Additionally, cell proliferation was significantly reduced and cell cycle was arrested in G1 phase after treatment with demethylating agents. Conclusions: Taken together, our results suggest that NDRG2 is a candidate tumor suppressor gene in OTs whose inactivation could be involved in tumoral progression and worse patient survival. The microarray study involved 28 glioma samples including oligodendrogliomas and oligoastrocytomas WHO grade II and III. Six non-tumoral brain tissues were used as controls, two of which were purchased from Stratagene (La Jolla, CA) and Clontech (Mountain View, CA). RNAs from several cell lines (Universal Human RNA, Stratagene, La Jolla, CA) was used as a standard reference in all hybridizations