Vascular endothelium forms a continuous, semipermeable barrier that regulates the transvascular movement of hormones, macromolecules, and other solutes. Here, we describe a novel immediate early gene that is expressed selectively in vascular endothelial cells, verge (vascular early response gene). Verge protein includes an N-terminal region of ∼70 amino acids with modest homology (∼30% identity) to Apolipoprotein L but is otherwise unique. Verge mRNA and protein are induced selectively in the endothelium of adult vasculature by electrical or chemical seizures. Verge expression appears to be responsive to local tissue conditions, because it is induced in the hemisphere ipsilateral to transient focal cerebral ischemia. In contrast to the transient expression in adult, Verge mRNA and protein are constitutively expressed at high levels in the endothelium of developing tissues (particularly heart) in association with angiogenesis. Verge mRNA is induced in cultured endothelial cells by defined growth factors and hypoxia. Verge protein is dramatically increased by cysteine proteinase inhibitors, suggesting rapid turnover, and is localized to focal regions near the periphery of the cells. Endothelial cell lines that stably express Verge form monolayers that show enhanced permeability in response to activation of protein kinase C by phorbol esters. This response is accompanied by reorganization of the actin cytoskeleton and the formation of paracellular gaps. These studies suggest that Verge functions as a dynamic regulator of endothelial cell signaling and vascular function.
Granulocyte-macrophage colony stimulating factor (GM-CSF) is a hematopoietic growth factor involved in the generation of granulocytes, macrophages, and dendritic cells from hematopoietic progenitor cells. We have recently demonstrated that GM-CSF has anti-apoptotic functions on neurons, and is neuroprotective in animal stroke models.The GM-CSF receptor alpha is expressed on adult neural stem cells in the rodent brain, and in culture. Addition of GM-CSF to NSCs in vitro increased neuronal differentiation in a dose-dependent manner as determined by quantitative PCR, reporter gene assays, and FACS analysis.Similar to the hematopoietic factor Granulocyte-colony stimulating factor (G-CSF), GM-CSF stimulates neuronal differentiation of adult NSCs. These data highlight the astonishingly similar functions of major hematopoietic factors in the brain, and raise the clinical attractiveness of GM-CSF as a novel drug for neurological disorders.
Granulocyte-colony stimulating factor (G-CSF) is a potent hematopoietic factor that drives differentiation of neutrophilic granulocytes. We have recently shown that G-CSF also acts as a neuronal growth factor, protects neurons in vitro and in vivo, and has regenerative potential in various neurological disease models. Spinal cord injury (SCI) following trauma or secondary to skeletal instability is a terrible condition with no effective therapies available at present. In this study, we show that the G-CSF receptor is up-regulated upon experimental SCI and that G-CSF improves functional outcome in a partial dissection model of SCI. G-CSF significantly decreases apoptosis in an experimental partial spinal transsection model in the mouse and increases expression of the anti-apoptotic G-CSF target gene Bcl-X(L). In vitro, G-CSF enhances neurite outgrowth and branching capacity of hippocampal neurons. In vivo, G-CSF treatment results in improved functional connectivity of the injured spinal cord as measured by Mn(2+)-enhanced MRI. G-CSF also increased length of the dorsal corticospinal tract and density of serotonergic fibers cranial to the lesion center. Mice treated systemically with G-CSF as well as transgenic mice over-expressing G-CSF in the CNS exhibit a strong improvement in functional outcome as measured by the BBB score and gridwalk analysis. We show that G-CSF improves outcome after experimental SCI by counteracting apoptosis, and enhancing connectivity in the injured spinal cord. We conclude that G-CSF constitutes a promising and feasible new therapy option for SCI.
Eine optimale Darmvorbereitung ist für die Durchführung einer validen koloskopischen Untersuchung essenziell. 2 Liter-PEG + Ascorbat (Moviprep®) wird seit 2006 in Deutschland vertrieben und hat in kontrollierten Studien dem konventionellen 4-Liter-PEG-Regime vergleichbare Ergebnisse gezeigt.
Background: Imaging based selection of acute ischemic stroke patients may improve clinical outcomes after endovascular therapy. DEFUSE 2 was a prospective cohort study of ischemic stroke patients who underwent a baseline MRI scan before endovascular therapy. In DEFUSE 2 reperfusion was associated with increased good functional outcome in patients with the target mismatch (TMM) profile, but not in patients without TMM. AXIS 2 was a randomized, controlled, MRI based trial of IV GM-CSF versus placebo. This study, in which endovascular therapy was not permitted, did not show improved functional outcomes between GM-CSF and placebo. We performed an indirect comparison between the outcomes seen in DEFUSE 2 and a control group from AXIS 2. Methods: Patients from AXIS 2 with a confirmed large vessel occlusion of the MCA or ICA on MR angiography, obtained within 9 hours after symptom onset, were selected as a control-group for patients from DEFUSE 2. The primary endpoint was good functional outcome at day 90 defined as a modified Rankin scale score of 0-2. Reperfusion status could only be assessed in the DEFUSE 2 cohort. We performed a stratified analysis based on the presence of TMM for both studies and reperfusion status in DEFUSE 2. Results: We compared good functional outcome in 102 patients (TMM in n=47, 49%) from AXIS 2 and 98 patients (TMM in n=78, 80%) from DEFUSE 2. After correction for differences in age, NIHSS and DWI volume, rates of good functional outcome were similar, 43% in DEFUSE 2 compared to 30 % in AXIS 2 (OR 0.9, 95%CI 0.5-1.7). In DEFUSE 2 patients with the TMM profile in whom reperfusion was achieved, the rate of good functional outcome was increased compared to TMM patients in AXIS 2, 64% versus 32% (OR 3.2, 95%CI 1.2-8.4). In TMM patients from DEFUSE 2 without reperfusion, the rate of good functional outcome was similar (31%), to the TMM patients in AXIS 2 (OR 1.0, 95%CI 0.4-2.3). No difference in outcomes was observed regardless of reperfusion status when a TMM was not present. Conclusion: This indirect, controlled comparison suggests that endovascular treatment resulting in reperfusion may lead to improved outcomes in patients with the TMM profile.
The jimpy rumpshaker (jprsh) mutation is an amino acid substitution in exon 4 (Ile186-->Thr) of the proteolipid protein (PLP) gene on the X chromosome. Affected mice show moderate hypomyelination of the central nervous system (CNS) with increased numbers of oligodendrocytes in the white matter of the spinal cord, a feature distinguishing them from other PLP mutations such as jp, in which premature cell death occurs with reduced numbers of oligodendrocytes. Myelin sheaths of jprsh immunostain for myelin basic protein (MBP) and DM-20, but very few contain PLP. This study examines the differentiation of oligodendrocytes cultured from the spinal cords of young mutant and wild type mice using various surface and cytoplasmic antigenic markers to define the stage of development. The majority of oligodendrocytes from mutant mice progress normally to express MBP; approximately 30%, relative to wild type, contain DM-20 at the in vivo age of 16 days, but very few immunostain for PLP or the O10 and O11 markers. The morphology of mutant cells in respect to membrane sheets and processes appears similar to normal. The jprsh oligodendrocyte is, therefore, characterized by a failure to express the markers indicative of the most mature cell; however, it is probably able to achieve a normal period of survival. These data, taken in conjunction with previous results, suggest that the PLP gene has at least two functions; one, probably involving PLP, is concerned with a structural role in normal myelin compaction; the other, perhaps related to DM-20 (or another lower molecular weight proteolipid), is essential for cell survival. The mutation in jprsh at residue 186 suggests that this region, which is common to PLP and DM-20, is not critical for this latter function.
OBJECTIVE: Accelerated treatment development for ALS patients needs validated biomarkers. G-CSF seems to compensate rapid neuronal loss in active neurodegeneration. Compassionate use of G-CSF in ALS patients may assist in long-term follow up of patients and potential biomarkers. BACKGROUND: With wide pathogenic heterogeneity of ALS, measurement of pyramidal tract integrity by DWI-MRI (Fractional Anisotropy, FAI) and of motor neuron integrity by neurophysiology (MUNE / MUNIX, e.g. hypothenar muscle) as downstream markers of potential therapy effects would be desirable biomarkers. MUNIX correlates well with disease progression in ALS, being more sensitive than changes of ALS-FRS-r. DWI-MRI studies in ALS patients have already demonstrated sensitivity to disease progression. Hematopoietic bone marrow stem cell differentiation markers may serve as additional biomarkers and help understand G-CSF`s mode of action. DESIGN/METHODS: 23 ALS patients (15 male, 8 female, mean age 51, 4 yrs.) received individual G-CSF treatments in monthly outpatient regimens and exams for ALS-FRS-r (mean at start 36.75 (± 6.84)), hematology and clinical chemistry. As reporting biomarkers we assessed (1) upper motor neuron integrity (DWI-MRI) and (2) lower motor neuron function (i-MUNIX), every 3 months during long-term (> 4 yrs.) treatment. RESULTS: Disease progression correlated significantly (p<0,0001) with loss of lower motor neurons (i-MUNIX); there was a trend in correlating FAI with ALS-FRS-r decline. i-MUNIX was significantly more sensitive for disease progression than ALS-FRS-r. During disease progression patients with lower ALS-FRS-r mobilized less monocytes and CD34+38- stem cells, but more eosinophils. Colony Forming Units for bone marrow differentiation lines are further analyzed. CONCLUSIONS: i-MUNIX, DWI-MRI, and bone marrow differentiation parameters are very promising biomarkers for ALS-treatment development. Validating these biomarkers for disease progression and potential sensitivity of treatment efficacy estimation is ongoing.
