ABSTRACT: Our aim was to evaluate the expression of chemokines in breast cancers before treatment to identify a molecular signature that could distinguish between women with or without cancer. We identified a panel of chemokines clearly deregulated in breast cancer that could be further investigated for prospective novel markers or therapeutically approaches. Samples for gene expression were obtained during surgical resection and after macroscopic pathological assessment. Laser-capture microdissection was used to select and procure only the desired cell types (malignant groups of cells or normal mammary acini), under direct microscopic visualization Chemokine expression profiles using cDNA microarray was evaluated on homogenous, laser-capture microdissected breast cancer specimens before treatment.
Project description:Our aim was to evaluate the expression of chemokines in breast cancers before treatment to identify a molecular signature that could distinguish between women with or without cancer. We identified a panel of chemokines clearly deregulated in breast cancer that could be further investigated for prospective novel markers or therapeutically approaches. Samples for gene expression were obtained during surgical resection and after macroscopic pathological assessment. Laser-capture microdissection was used to select and procure only the desired cell types (malignant groups of cells or normal mammary acini), under direct microscopic visualization
Project description:Pancreatic ductal cells and preneoplastic lesions derived from wild type, Keratin 5 COX-2, P48Cretg/wt K-RasG12D/wt and K5 COX-2 P48Cretg/wt K-RasG12D/wt mouse genotypes were laser capture microdissected. Total RNA was isolated and analysed with Illumuna Sentrix 8 chips.
Project description:Expression profiling of a total of 566 miRNA genes and controls in 24 laser microdissected samples (normal lung parenchyma, normal mammary glands, lung metastases, primary tumor carcinoma and primary tumor EMT) and Jyg MC (A) GFP/Luc cells and spheres Laser-capture microdissection combined with NanoString gene expression assays of microRNAs revealed overexpression of either epithelial or mesenchymal markers in epithelial and spindle-like cell regions, respectively
Project description:The continuously remodeled extracellular matrix (ECM) plays a pivotal role in gastrointestinal health and disease, yet its precise functions remain elusive. In this project, we employed laser capture microdissection of cecal mucosa combined with low-input proteomics to investigate ECM remodeling during Salmonella-driven inflammation. To complement this, we probed how fibronectin fiber tension is altered using a mechanosensitive peptide probe. While fibronectin fibers in healthy intestinal tissue are typically stretched, fibronectin fiber relaxation occurred exclusively during late-stage infection at 72 hours and was localized to already existing clusters of infiltrated neutrophils in the cecal mucosa of Salmonella-infected mice.
Project description:We provide an original multi-stage approach identifying a gene signature to assess the fibroblast polarization. Prototypic polarizations (inflammatory/fibrotic) were induced by seeded mouse embryonic fibroblasts (MEFs) with TNFα or TGFß1, respectively. The transcriptomic and proteomic profiles were obtained by RNA microarray and LC/MS-MS. Gene Ontology and pathways analysis were performed among the differentially expressed genes (DEGs) and proteins (DEPs). Balb/c mice underwent daily intradermal injections of HOCl (or PBS) as an experimental murine model of inflammation-mediated fibrosis in a time-dependent manner. As results, 1,456 and 2,215 DEGs, and 289 and 233 DEPs were respectively found in MEFs in response to TNFα or TGFß1, respectively. Among the most significant pathways, we combined 26 representative genes to encompass the proinflammatory and profibrotic polarizations of fibroblasts. Based on principal component analysis, this signature deciphered baseline state, proinflammatory polarization, and profibrotic polarization as accurately as did RNA microarray and LC/MS-MS. Then, we assessed the gene signature on dermal fibroblasts isolated from the experimental murine model. We observed a proinflammatory polarization at day 7, and a mixture of a proinflammatory and profibrotic polarizations at day 42 in line with histological findings. Our approach provides a small-size and convenient gene signature to assess murine fibroblast polarization.
Project description:The purpose of this study was to identify differentially expressed genes in laser-capture microdissected (LCM) invasive mammary carcinomas (IMCs). Invasive mammary carcinoma of the breast surgical resection specimens were laser capture microdissected for RNA extraction and hybridization to Affymetrix microarrays.
Project description:We analysed gene expression in normal breast blood vessels and breast invasive ductal carinoma blood vessels. Normal breast sections and IDC breast sections were immunostained for endothelial-specific CD31. Vessels were dissected and captured with laser capture microscopy, and RNA was extracted and analysed on an Affymetrx array chip. In total, gene expression in blood vessels from 3 normal breast and 3 IDC breast was analysed.
Project description:Cortical interneurons originating from the medial ganglionic eminence (MGE) are among the most diverse cells within the CNS. Different pools of proliferating progenitor cells are thought to exist in the ventricular zone of the MGE, but whether the underlying subventricular and mantle regions of the MGE are spatially patterned has not yet been addressed. Here, we combined laser-capture microdissection and multiplex RNA-sequencing to map the transcriptome of MGE cells at a spatial resolution of 50 M-BM-5m. Distinct groups of progenitor cells showing different stages of interneuron maturation were identified and topographically mapped based on their genome-wide transcriptional pattern. One 50 M-BM-5m coronal section from the MGE was taken from each of two wildtype and one GFRa1 mutant E12.5 C57bl6/J mouse. Each section was laser microdissected into approximately 100 cubes, covering the whole MGE, and each cube was further processed for RNA-seq analysis.
Project description:Tumor-associated breast stroma was laser-capture microdissected from IDC breast cancer cases. The goal of the study was to characterize the heterogeneity of breast tumor-assocaited stroma and identify gene expression signatures predictive of clinical outcome. Experiment Overall Design: Common reference design, 53 samples, with dye-swap replicates. Some samples replicated three or four times, a total of 111 arrays.