Aileen Healy

Mice deficient in PKC theta demonstrate impaired in vivo T cell activation and protection from T cell-mediated inflammatory diseases.

In the present study we have characterized T cell-driven immune function in mice that are genetically deficient in PKC theta. In response to simple immunologic stimulation invoked by in vivo T cell receptor (TCR) cross-linking, these mice showed significantly depressed plasma cytokine levels for IL-2, IL-4, IFNγ, and TNFα compared to wild-type (WT) mice. In parallel, spleen mRNA levels for these cytokines were reduced, and NF-κB activation was also reduced in PKC theta knockouts (KO). Injection of allogeneic cells into the footpad of PKC theta deficient mice provoked a significantly diminished local T cell response compared to WT mice similarly challenged. Unlike comparable cells from wild type mice, CD45RBhi T cells harvested from PKC theta deficient mice failed to induce colitis in the SCID-CD45RB cell transfer model of IBD. In another T cell-dependent model of inflammatory disease, PKC theta deficient animals developed far less severe neurologic signs and reduced spinal cord inflammatory cell infiltrate compared to WT controls in the MOG-induced EAE model. A fundamental role for PKC theta in T cell activation and in the development of T cell-mediated inflammatory diseases is indicated by these results.

Fragile X syndrome: an update on developing treatment modalities.

Intellectual disability (ID; mental retardation) is considered an immutable condition. Current medical practices are aimed at relieving symptoms and not at altering the underlying cognitive deficits. Scientific advancements from the past decade have led to the exciting possibility that ID may now be treatable. Moreover, pharmaceutical therapies targeting the most common form of inherited ID, Fragile X syndrome (FXS), may become the new benchmark for central nervous system (CNS) drug discovery: seeking cures for neurodevelopmental disorders.

Quantitation of peripheral blood markers of rat experimental autoimmune encephalomyelitis.

Identification and quantitation of peripheral blood non-invasive, cell-surface markers of EAE disease activity and drug response would facilitate the preclinical development of potential therapeutics. Towards this end, we characterized the influx of immune mediators into spinal cords of diseased rats to establish the kinetics of T cell and monocyte-mediated inflammation. We then examined the periphery for regulation of T cell and monocyte activation. We report increased CD80 and VLA-4 expression on peripheral blood monocytes (PBM) during the onset and peak of experimental disease scores. Increased CD4+, CD62L − and CD4+, CD134+ T cells were detected only at disease peak, not during disease onset. PBM CD80 expression was significantly inhibited in CSA-treated animals, but increased in Dex-treated animals. PBM VLA-4 expression was unaffected by drug treatment. Both CSA and Dex inhibited CD62L shedding and CD134 expression on peripheral CD4+ T cells. These results identify quantitative, peripheral markers of disease activity and drug response.

Identificação de genes envolvidos na síntese de proteínas de células musculares lisas com expressão aumentada em placas ateromatosas associados a hiperplasia neointimal após implante de stents não-farmacológicos: estudo GENESIS-R

BACKGROUND: Coronary restenosis is a poorly understood phenomenon that remains a challenge even in the drug-eluting stent era. This study is aimed at identifying the genes involved in structural and functional protein synthesis of smooth muscle cells with increased expression in human atheromatous plaques associated to neointimal hyperplasia after bare-metal stent implantation. METHODS: Atheromatous plaques were obtained by directional atherectomy prior to stenting. Gene expression analysis was performed using the Affymetrix GeneChip system. Patients were submitted to intravascular ultrasound 6 months after the procedure for in-stent volumetric analysis. We evaluated the correlation between gene expression in atheromatous plaques and the percentage of in-stent intimal hyperplasia. RESULTS: Most patients were male (85.7%), with 60.2 ± 11.4 years of age, 35.7% were diabetic and the percentage of in-stent intimal hyperplasia was 29.9 ± 18.7%. There was no change in the percentage of in-stent intimal hyperplasia in patients with or without diabetes (29.5% vs. 30.7%; P = 0.89). There was no correlation between stent length and the percentage of in-stent intimal hyperplasia (r = -0.26; P = 0.26) or between stent diameter and the percentage of in-stent intimal hyperplasia (r = 0.14; P = 0.56). Eight genes related to smooth muscle cell structural and functional protein synthesis had a positive correlation with the percentage of in-stent intimal hyperplasia. CONCLUSIONS: De novo coronary lesions show increased expression of genes related to smooth muscle cell structural and functional protein synthesis associated to future significant in-stent neointimal hyperplasia, emerging as novel therapeutic targets.

Response to Letter Regarding Article, “Platelet Expression Profiling and Clinical Validation of Myeloid-Related Protein-14 as a Novel Determinant of Cardiovascular Events”

BACKGROUND Platelets participate in events that immediately precede acute myocardial infarction. Because platelets lack nuclear DNA but retain megakaryocyte-derived mRNAs, the platelet transcriptome provides a novel window on gene expression preceding acute coronary events. METHODS AND RESULTS We profiled platelet mRNA from patients with acute ST-segment-elevation myocardial infarction (STEMI, n=16) or stable coronary artery disease (n=44). The platelet transcriptomes were analyzed and single-gene models constructed to identify candidate genes with differential expression. We validated 1 candidate gene product by performing a prospective, nested case-control study (n=255 case-control pairs) among apparently healthy women to assess the risk of future cardiovascular events (nonfatal myocardial infarction, nonfatal stroke, and cardiovascular death) associated with baseline plasma levels of the candidate protein. Platelets isolated from STEMI and coronary artery disease patients contained 54 differentially expressed transcripts. The strongest discriminators of STEMI in the microarrays were CD69 (odds ratio 6.2, P<0.001) and myeloid-related protein-14 (MRP-14; odds ratio 3.3, P=0.002). Plasma levels of MRP-8/14 heterodimer were higher in STEMI patients (17.0 versus 8.0 microg/mL, P<0.001). In the validation study, the risk of a first cardiovascular event increased with each increasing quartile of MRP-8/14 (Ptrend<0.001) such that women with the highest levels had a 3.8-fold increase in risk of any vascular event (P<0.001). Risks were independent of standard risk factors and C-reactive protein. CONCLUSIONS The platelet transcriptome reveals quantitative differences between acute and stable coronary artery disease. MRP-14 expression increases before STEMI, and increasing plasma concentrations of MRP-8/14 among healthy individuals predict the risk of future cardiovascular events.