Project description:Targeting of tumor immune escape mechanisms holds enormous therapeutic potential. Still, most patients progress under immune checkpoint blockade and some even become hyper-progressors. To investigate how cancer cells respond to activated but ineffective T cells, we challenged peptide-loaded MCF-7 breast cancer cells with antigen-specific CD8+ T cells in which Perforin had been destroyed by pre-treatment with Concanamycin A.
Project description:To investigate the difference in gene expression of liver cancer cells after co-culture with activated CD8+T cells , we established a tumor cells/activated CD8+T cells co-culture system in which murine liver cancer cell line hepa1-6 were co-cultured and attacked directly by murine in vitro activated spleen naive CD8+T cells (effector: target=5:1) for 48 hours. We then performed gene expression profiling analysis using data obtained from RNA-seq of two different groups (immune attacked or control) with 5 repetitions per group.
Project description:<p>Breast cancer metastasis occurs via blood and lymphatic vessels. Breast cancer cells 'educate' lymphatic endothelial cells (LECs) to support tumor vascularization and growth. However, despite known metabolic alterations in breast cancer, it remains unclear how lymphatic endothelial cell metabolism is altered in the tumor microenvironment and its effect in lymphangiogenic signaling in LECs. We analyzed metabolites inside LECs in co-culture with MCF-7, MDA-MB-231, and SK-BR-3 breast cancer cell lines using 1H nuclear magnetic resonance (NMR) metabolomics, Seahorse, and the spatial distribution of metabolic co-enzymes using optical redox ratio imaging to describe breast cancer-LEC metabolic crosstalk. LECs co-cultured with breast cancer cells exhibited cell-line dependent altered metabolic profiles, including significant changes in lactate concentration in breast cancer co-culture. Cell metabolic phenotype analysis using Seahorse showed LECs in co-culture exhibited reduced mitochondrial respiration, increased reliance on glycolysis and reduced metabolic flexibility. Optical redox ratio measurements revealed reduced NAD(P)H levels in LECs potentially due to increased NAD(P)H utilization to maintain redox homeostasis. 13C-labeled glucose experiments did not reveal lactate shuttling into LECs from breast cancer cells, yet showed other 13C signals in LECs suggesting internalized metabolites and metabolic exchange between the two cell types. We also determined that breast cancer co-culture stimulated lymphangiogenic signaling in LECs, yet activation was not stimulated by lactate alone. Increased lymphangiogenic signaling suggests paracrine signaling between LECs and breast cancer cells which could have a pro-metastatic role.</p>
Project description:The persistence of HIV infection under ART is due to a reservoir of latently infected cells that harbor replication-competent virus and evade immune recognition. Defining the mechanisms responsible for the establishment and maintenance of HIV latency is crucial to achieve HIV eradication or functional cure. Previous studies demonstrated a non-cytotoxic CD8+ T-cells mediated inhibition of virus replication during untreated HIV/SIV infection and inhibition of virus production under ART; however, the mechanisms responsible for this antiviral effect remained poorly understood. In our primary cell-based in vitro latency model we demonstrated that co-culture with CD8+ T-cells promotes changes in metabolic and cell survival pathways in HIV-infected memory CD4+ T-cells that may negatively regulate HIV expression and ultimately promote the establishment of latency. Modulation of this CD8-mediated activity may represent a tool to disrupt HIV latency and reservoir persistence in ART-treated individuals.
Project description:Aerobic glycolysis is a hallmark of cancer glucose metabolism. Here we suggest that extracellular vesicles (EVs) originating from cancer cells can modulate glucose metabolism in the recipient cancer cells and induce cell proliferation and aggressive cancer phenotypes. Two breast cancer cell lines with different levels of glycolytic activity, MDA-MB-231 and MCF7, were selected and co-cultured, as the originating and recipient cells. The change in 18F-fluorodeoxyglucose (FDG) uptake of the recipient MCF7 cells was assessed after co-culture with the MDA-MB-231 cells. Proteomics analysis was performed to investigate the changes in the protein expression patterns in the recipient MCF7 cells. FDG uptake by the recipient MCF7 cells was sig-nificantly increased after co-culture with the MDA-MB-231 cells.
Project description:Three-dimensional (3D) cell direct co-culture is an essential method for cancer research, which can overcome the limitations of traditional research based on cell lines. Single-cell RNA sequencing can detect the cell heterogeneity of direct co-culture of 3D cells at the single-cell level,However, there is still lack verification. The purpose of this study is to verify the superiority of single-cell RNA sequencing combined with 3D cell direct co-culture in breast cancer research and prove that it can become a powerful solution for cancer cell culture in vitro.In this study ,we found that the different cell culture modes showed very different responses to cisplatin intervention. In MCF-7 alone culture mode, there was a large difference in the transcriptome of MCF-7 after cisplatin intervention compared with the group not given drug intervention; while in direct co-culture mode, there was also a difference between the cisplatin intervention group and the control group, but this difference was relatively small. To further seek evidence that the direct co-culture mode is closer to the real environment in vivo, we analyzed the differential genes in the direct co-culture mode, and the enriched signaling pathways were similar to the results in the CancerSEA database, and the enriched signaling pathways included apoptosis, DNA damage, hypoxia, and metastasis. Conclusions: Single-cell sequencing can facilitate direct co-culture of 3D cells in tumor research, a model that more closely resembles the real pathophysiological environment in vivo.