Project description:Idelalisib has excellent activity in management of chronic lymphocytic leukemia and follicular lymphoma while inducing serious microbial pneumonia in the clinic. To understand the mechanisms of Idelalisib-induced infection risk, we performed RNAseq analyses on mouse precision-cut lung slices treated with Idelalisib in the absence and presence of human lung pathogen Streptococcus pneumoniae. RNAseq transcriptome profiling identifies distinct response signatures in mouse lung slices in responses to Idelalisib treatment and pneumococcal infection. The expression of genes vital for regulation of transcription, immunity and inflammation, cell proliferation and apoptosis, kinase signaling, metabolism, and extracellular matrix remodeling is significantly modulated by Idelalisib. This study provides a new insight for predicting the immunomodulatory impact of drug candidates on host defense and thereby improving the safety profiling of novel drugs for human respiratory diseases.
Project description:Purpose: To identify cell types and cellular transcriptional profiles in airway explants and precision cut lung slices obtained from non-disease human lung tissue.
Project description:This study was done to show the utility of precision-cut lung slices (PCLS) in supporting the survival of Pneumocystis murina in vitro.
Project description:We conducted a single cell RNA sequencing experiment to determine the effect of a macropinocytosis inhibitor (5-(N-ethyl-N-isopropyl)-Amiloride, EIPA) in Precision Cut Lung Slices.
Project description:The data provided here is related to the publication “Peptidomic and proteomic analysis of precision-cut lung slice supernatants”. In this study, we sought out to develop a method that could incorporate a peptidomic and proteomic endpoint within the same workflow. The developed workflow was adapted from single-pot, solid-phase-enhanced sample preparation (SP3) for examining the proteome and peptidome of precision-cut lung slice supernatants in one experimental procedure to save sample material. A part of the study compared potential differences when utilizing different organic solvents for binding peptides and proteins to magnetic beads for biological replicates of PCLS supernatant. In connection, the workflow robustness was also characterized by investigating technical replicates of the same biological PCLS supernatant sample.
Project description:Purpose: The aim of this study was to create a patient-derived slice model by combining cryopreservation technique with precision-cut slice culture method and explore its effectivity of predicting anti-cancer drug sensitivity in vitro. Methods: We prepared 0.3 mm thick tissue slices by a microtome and maintain its cell viability by cryopreservation technique. Slices were cultured individually in the presence or absence of regorafenib (REG) for 72 hours. Alterations in morphology and gene expression were assessed by histological and genetic analysis. Overall viability was also analyzed in tissue slices by CCK-8 quantification assay and fluorescent staining. Tissue morphology and cell viability could be evaluated to quantify drug effects. Results: Histological and genetic analysis showed that no significant alterations in morphology and gene expression were induced by vitrification‑based cryopreservation. The viability of warmed HCC tissues was up to 90% of the fresh tissues. The viability and proliferation could be retained for at least four days in filter culture system. The positive drug responses in precision-cut slice culture in vitro were evaluated by tissue morphology and cell viability. Conclusion: In summary, the successful application of precision-cut HCC slice culture combining cryopreservation technique in a systematic drug screen demonstrates the feasibility and utility of slice culture method for drug response.
Project description:Ex vivo human precision-cut lung slices (PCLS) were infected with human parainfluenza virus 3 (hPIV-3). We used microarry to explore the changes in gene expression between infected and non-infected, same-donor PCLS. We particularly analyzed changes in mRNA and miRNA expression levels linked to inflammation and antiviral immunity.
Project description:Tissue resident host responses to microbial infections in the respiratory tract are highly dynamic in space and time and rely on the interaction of a multitude of cell types. To model these multicellular responses reliably in cell culture, we compare here the global transcriptional antimicrobial response to infection with influenza A virus (IAV) in precision cut lung slices (PCLS), volume defined organ discs largely maintaining the cellular composition and 3D architecture of the donor lung. To permit a fair comparison of host responses in an isogenic background we first challenged mice in vivo and murine PCLS (mPCLS) and assess host transcriptomic changes by unbiased RNAseq. While core antiviral responses overlapped substantially, mPCLS lacked certain features—such as type II interferon expression—likely due to the absence of infiltrating immune cells responses. Importantly, when expanding our findings to immune experienced human precision cut lung slices (hPCLS), we find a much broader antiviral response after IAV challenge, including type I, II and III interferons, suggesting the presence of responsive tissue resident lymphocytes. To prove specificity of this response we infected hPCLS with Streptococcus pneumoniae. Ex vivo tissues responded with a distinct proinflammatory gene profile including IL1A, IL1B and IL17 expression. Blocking of IL-1β signaling partially inhibited the proinflammatory response, suggesting cellular crosstalk and a complex and specific antimicrobial reaction in this ex vivo model. In conclusion diversified tissue resident immune cell compartment distinguishes the human ex vivo model, making it an ideal system for microbiological and immunological research.
