Enter Your Search:
Results 1-2 (2)
Go to page number:
Select a Filter Below
Journal of immunology (Baltimore, Md. : 1950) (1)
PLoS ONE (1)
Hart, Traver (2)
Bolusani, Swetha (1)
Bridger, Joanna Mary (1)
Campbell, Daniel (1)
Chen, Caifu (1)
Garg, Ankit (1)
Grigoryev, Yevgeniy A. (1)
Head, Steven R. (1)
Kurian, Sunil M. (1)
Marcotte, Edward M. (1)
Nakorchevsky, Aleksey A. (1)
Salomon, Daniel. R (1)
Yates, John R. (1)
Zhao, Alice (1)
Year of Publication
microRNA regulation of molecular networks mapped by global microRNA, mRNA, and protein expression in activated T-lymphocytes
Grigoryev, Yevgeniy A.
Kurian, Sunil M.
Nakorchevsky, Aleksey A.
Head, Steven R.
Yates, John R.
Salomon, Daniel. R
Journal of immunology (Baltimore, Md. : 1950)
MicroRNAs (miRNAs) regulate specific immune mechanisms but their genome-wide regulation of T-lymphocyte activation is largely unknown. We performed a multidimensional functional genomics analysis to integrate genome-wide differential mRNA, miRNA, and protein expression as a function of human T-lymphocyte activation and time. We surveyed expression of 420 human miRNAs in parallel with genome-wide mRNA expression. We identified a unique signature of 71 differentially expressed miRNAs, 57 of which were previously not known as regulators of immune activation. The majority of miRNAs are upregulated, mRNA expression of these target genes is downregulated and this is a function of binding multiple miRNAs (combinatorial targeting). Our data reveal that consideration of this complex signature, rather than single miRNAs, is necessary to construct a full picture of miRNA-mediated regulation. Molecular network mapping of miRNA targets revealed the regulation of activation-induced immune signaling. In contrast, pathways populated by genes that are not miRNA targets are enriched for metabolism and biosynthesis. Finally, we specifically validated miR-155 (known) and miR-221 (novel in T-lymphocytes) using locked nucleic acid inhibitors. Inhibition of these 2 highly upregulated miRNAs in CD4+ T cells were shown to increase proliferation by removing suppression of 4 target genes linked to proliferation and survival. Thus, multiple lines of evidence link top functional networks directly to T-lymphocyte immunity underlining the value of mapping global gene, protein and miRNA expression.
Adaptive immunity; lymphocyte activation; functional genomics; microRNA; high-throughput genomics; molecular networks; T-lymphocytes; immune regulation
Human Cell Chips: Adapting DNA Microarray Spotting Technology to Cell-Based Imaging Assays
Marcotte, Edward M.
Bridger, Joanna Mary
Here we describe human spotted cell chips, a technology for determining cellular state across arrays of cells subjected to chemical or genetic perturbation. Cells are grown and treated under standard tissue culture conditions before being fixed and printed onto replicate glass slides, effectively decoupling the experimental conditions from the assay technique. Each slide is then probed using immunofluorescence or other optical reporter and assayed by automated microscopy. We show potential applications of the cell chip by assaying HeLa and A549 samples for changes in target protein abundance (of the dsRNA-activated protein kinase PKR), subcellular localization (nuclear translocation of NFκB) and activation state (phosphorylation of STAT1 and of the p38 and JNK stress kinases) in response to treatment by several chemical effectors (anisomycin, TNFα, and interferon), and we demonstrate scalability by printing a chip with ∼4,700 discrete samples of HeLa cells. Coupling this technology to high-throughput methods for culturing and treating cell lines could enable researchers to examine the impact of exogenous effectors on the same population of experimentally treated cells across multiple reporter targets potentially representing a variety of molecular systems, thus producing a highly multiplexed dataset with minimized experimental variance and at reduced reagent cost compared to alternative techniques. The ability to prepare and store chips also allows researchers to follow up on observations gleaned from initial screens with maximal repeatability.
Results 1-2 (2)
Go to page number:
Remove citation from clipboard
Add citation to clipboard
This will clear all selections from your clipboard. Do you wish proceed?
Clipboard is full! Please remove an item and try again.
PubMed Central Canada is a service of the
Canadian Institutes of Health Research
(CIHR) working in partnership with the National Research Council's
Canada Institute for Scientific and Technical Information
in cooperation with the
National Center for Biotechnology Information
U.S. National Library of Medicine
(NCBI/NLM). It includes content provided to the
PubMed Central International archive
by participating publishers.