Single cell tracking of expressed barcodes in cancer cell lines upon administration of FOLFOX or FOLFIRI at different time points
Ontology highlight
ABSTRACT:
INSTRUMENT(S): Illumina NovaSeq 6000
ORGANISM(S): Homo sapiens
SUBMITTER: Davide Cittaro
PROVIDER: E-MTAB-12951 | biostudies-arrayexpress |
REPOSITORIES: biostudies-arrayexpress
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Project description:This experiment is aimed at studying the effect of two different chemotherapies (FOLFOX and FOLFIRI) on colorectal cancer cell lines (HCT116 and HT29) at different time points. Two early time points (8h and 16h) are used to evaluate the acute response, while a late time point (two weeks, LT) is used to evaluate the long-term effect. After two week exposure, cells are grown without treatments for additional two weeks, to evaluate the transcriptomic of cells which survived the chemotherapy. A pool of the same cells has been independently profiled by scGET-seq.
2023-09-01 | E-MTAB-12949 | biostudies-arrayexpress
Project description:This experiment is aimed at studying the effect of two different chemotherapies (FOLFOX and FOLFIRI) on gene expression of HCT116 colorectal cancer cell line at different time points. Two early time points (8h and 16h) are used to evaluate the acute response, while a late time point (two weeks, LT) is used to evaluate the long-term effect. After two week exposure, cells are grown without treatments for additional two weeks, to evaluate the transcriptomic of cells which survived the chemotherapy. For all time points, untreated cells (CTRL) are also collected.
2023-09-01 | E-MTAB-12950 | biostudies-arrayexpress
Project description:This experiment is aimed at studying the effect of two different chemotherapies (FOLFOX and FOLFIRI) on chromatin accessibility of HCT116 colorectal cancer cell line at different time points. Two early time points (8h and 16h) are used to evaluate the acute response, while a late time point (two weeks, LT) is used to evaluate the long-term effect. After two week exposure, cells are grown without treatments for additional two weeks, to evaluate the chromatin landscapes of cells which survived the chemotherapy. A pool of the same cells has been independently profiled by scRNA-seq.
2023-09-01 | E-MTAB-12952 | biostudies-arrayexpress
Project description:This experiment is aimed at studying the effect of two different chemotherapies (FOLFOX and FOLFIRI) on chromatin accessibility of HCT116 colorectal cancer cell line at different time points. Two early time points (8h and 16h) are used to evaluate the acute response, while a late time point (two weeks, LT) is used to evaluate the long-term effect. After two week exposure, cells are grown without treatments for additional two weeks, to evaluate the chromatin landscapes of cells which survived the chemotherapy. A pool of the same cells has been independently profiled by scRNA-seq.
2023-09-01 | E-MTAB-12953 | biostudies-arrayexpress
Project description:This experiment is aimed at studying the effect of two different chemotherapies (FOLFOX and FOLFIRI) on colorectal cancer cell lines (HCT116 and HT29) at different time points. Two early time points (8h and 16h) are used to evaluate the acute response, while a late time point (two weeks, LT) is used to evaluate the long-term effect.
2023-09-01 | E-MTAB-12948 | biostudies-arrayexpress
Project description:This experiment is aimed at studying the effect of two different chemotherapies (FOLFOX and FOLFIRI) and their interaction with a ULK1 inhibitor on HCT116 colorectal cancer cell line.
2023-09-01 | E-MTAB-12947 | biostudies-arrayexpress
Project description:The aim of the experiment is to elucidate changes in tumor heterogeneity upon chemotherapy treatment. The experiment includes non-treated samples, samples right after chemotherapy treatment (FOLFIRI) and organoids recovered from FOLFIRI. It was conducted in two different genetic backgrounds, AKP (Apc KO, KrasG12D and P53 KO) and APS (Apc KO, P53 and Smad4KO). In addition to that, the organoids carry an inducible Cre-ERT2 knock-in allele in the mex3a locus. Upon 4-OH-Tamoxifen induction, Mex3a cells will recombine the tracing allele stop-TdTomato, enabling the tracing of this subpopulation. To understand the fate of the traced cells, samples right after chemotherapy and in the recovery setting were sorted based on their tomato expression. This enables a single cell tracing experiment.
2022-05-13 | E-MTAB-11145 | biostudies-arrayexpress
Project description:Mex3a is an RNA binding protein of unknown function. To elucidate the contribution of Mex3a to tumoral heterogeneity, Mex3a KO organoids engineered by CRISPR were sequenced in three different conditions. Live organoids (DAPI negative) were sorted in Control, after 2 days of FOLFIRI and after 5 days of treatment. Two WT organoids (parental and a derived clone) and two KO (KO1 and KO2, two independent clones) were used for this experiment.
2022-05-16 | E-MTAB-11146 | biostudies-arrayexpress
Project description:In patients with advanced colorectal cancer, leucovorin, fluorouracil, and irinotecan (FOLFIRI) is considered as one of the reference first-line treatments. However, only about half of treated patients respond to this regimen, and there is no clinically useful marker that predicts response. A major clinical challenge is to identify the subset of patients who could benefit from this chemotherapy. We aimed to identify a gene expression profile in primary colon cancer tissue that could predict chemotherapy response. Patients and Methods:- Tumor colon samples from 21 patients with advanced colorectal cancer were analyzed for gene expression profiling using Human Genome GeneChip arrays U133. At the end of the first-line treatment, the best observed response, according to WHO criteria, was used to define the responders and nonresponders. Discriminatory genes were first selected by the significance analysis of microarrays algorithm and the area under the receiver operating characteristic curve. A predictor classifier was then constructed using support vector machines. Finally, leave-one-out cross validation was used to estimate the performance and the accuracy of the output class prediction rule. Results:- We determined a set of 14 predictor genes of response to FOLFIRI. Nine of nine responders (100% specificity) and 11 of 12 nonresponders (92% sensitivity) were classified correctly, for an overall accuracy of 95%. Conclusion:- After validation in an independent cohort of patients, our gene signature could be used as a decision tool to assist oncologists in selecting colorectal cancer patients who could benefit from FOLFIRI chemotherapy, both in the adjuvant and the first-line metastatic setting. All tissue samples were maintained at −180°C (liquid nitrogen) until RNA extraction and were weighed before homogenization. Tissue samples were then disrupted directly into a lysis buffer using Mixer Mill MM 300 (Qiagen, Valencia, CA). Total RNA was isolated from tissue lysates using the RNeasy Mini Kit (Qiagen), and additional DNAse digestion was performed on all samples during the extraction process (RNase-Free DNase Set Protocol for DNase treatment on RNeasy Mini Spin Columns; Qiagen). After each extraction, a small fraction of the total RNA preparation was taken to determine the quality of the sample and the yield of total RNA. Controls analyses were performed by UV spectroscopy and analysis of total RNA profile using the Agilent RNA 6000 Nano LabChip Kit with the Agilent 2100 Bioanalyser (Agilent Technologies, Palo Alto, CA) to determine RNA purity, quantity, and integrity.
2014-10-06 | E-GEOD-62080 | biostudies-arrayexpress
Project description:Purpose of the study: the goal of this study was to use Next-generation sequencing to investigate the gene expression profile of skeletal muscle from mice exposed to chemotherapy (Folfiri) for up to 5 weeks. Methods: Standard methods were used for polyA mRNA-seq library construction, EZBead preparation and Next-Gen sequencing, based on Life Technologies SOLiD5000xl system. Briefly, one microgram total RNA per sample was applied for library preparation. PolyA mRNA was first captured using the standard protocol of Dynabeads® mRNA DIRECT⢠Micro Kit (Life Technologies, #61021). Following the enrichment of polyA mRNA, the cDNA library was prepared and barcoded per sample using the standard protocol of SOLiD Total RNA-seq Kit (Life Technologies, #4445374). Each barcoded library was quantified by Bioanalyzer High Sensitivity DNA chip (Agilent, #5067-4626) and pooled in equal molarity. EZBead preparation, bead library amplification, and bead enrichment were then conducted using Life Technologies EZ Bead⢠E80 System (Life technologies, #4453095). Approximately four hundred forty million library-enriched beads per lane were deposited onto a SOLiD5500xl FlowChip (6 lanes/chip). Finally sequencing by ligation was carried out using standard single-read, 5â-3â strand-specific sequencing procedure (75b-read) on SOLiD5500xl Sequencer. The resulting 75 bp solid reads were mapped to Mus musculus mm9 reference genome using in-house mapping pipelines that utilizes bfast-0.7.0a [67]. In brief, using our RNA-seq pipeline, low quality reads and reads mapped to rRNA/tRNAs were first discarded. The remaining reads were mapped to reference genome mm9 and a splice-junction library, respectively; the genomic and splice-junction library mapping were merged at the end. The gene based expression levels were calculated using bamutils from NGSUtils based on the RefSeq gene annotation of mm9. Differential expression of genes across different treatments was determined with edgeR Results: Gene expression profiling, performed by means of RNA-Sequencing analysis, identified a limited number of genes that were significantly modulated (False Discovery Rate < 0.05) following Folfiri administration. Interestingly, the pathway analysis showed marked down-regulation of the regulators of mitochondrial metabolism Ucp1, Cidea1 and Acot2, as well as the marker of muscle cell proliferation/pluripotency Fhl3 . Further, we found increased expression of regulators of lipid metabolism and transport (such as Fabp1, Apoa1, Apob, Apoa2, Alb, Prkcz and Scd2) as well as acute phase response proteins (such as Alb, Fga and Fgb), and members of the PPAR signaling and markers of the Energy metabolism (such as Dnah5) were reported . Conclusions: Our study represents the first gene expression profiling of muscle from mice exposed to chemotherapy. The findings from our study identified several signaling pathways that were modulated following chemotherapy administration. These results will contribute to identify new modulators of muscle wasting associated with Folfiri administration, that might also represent putative new targets of interventions. Next-generation sequencing of RNA extracted from quadriceps muscle of CD2F1 male mice exposed to either Folfiri (a combination of 5-FU, Leucovorin and CPT-11) or vehicle alone (n=4) for up to 5 weeks.
2016-05-18 | E-GEOD-80473 | biostudies-arrayexpress