Sven Wellmann

Hypoxia upregulates the histone demethylase JMJD1A via HIF-1

The histone demethylase Jumonji domain containing 1A (JMJD1A) demethylates H3K9 residues and thereby transactivates distinct target genes. Investigating the effect of hypoxia on JMJD1A expression, we found increased JMJD1A mRNA in different organs of rats exposed to normobaric hypoxia (8% O(2)). Compared to adult samples, JMJD1A was increased in most tissues of human fetuses in whom oxygen supply is low compared to postnatal levels. Upregulation of JMJD1A mRNA and protein in cultured human cells exposed to hypoxia or iron scavengers in vitro was abrogated when hypoxia-inducible factor-1 (HIF-1) signaling was blocked by siRNAs. A single pivotal hypoxia responsive element (HRE) in the promoter of the human JMJD1A gene was identified that mediates JMJD1A upregulation by hypoxia, iron scavengers, and HIF-1. These findings demonstrate that JMJD1A can be stimulated by hypoxia both in vitro and in vivo involving binding of HIF-1 to a specific HRE in the JMJD1A promoter.

Subfield-specific neurovascular remodeling in the entorhino-hippocampal-organotypic slice culture as a response to oxygen-glucose deprivation and excitotoxic cell death

Transient ischemia causes delayed neurodegeneration in selective brain areas, particularly in the CA1 field of the hippocampus. This is accompanied by neurovascular impairment. It is unknown whether neurodegeneration is the cause or consequence of vascular changes. In an entorhino-hippocampal-organotypic slice culture system with well-preserved blood vessels, we studied the interplay between neurodegeneration and neurovasculature. Short-term oxygen and glucose deprivation (OGD) resulted in upregulation of hypoxic markers and with a delay of 24 to 48 hours in selective nerve cell death in CA1. In parallel, local vessel density decreased as detected by markers of endothelial cells and of the extracellular matrix. Claudin-5, a tight junction protein and marker of the blood–brain barrier was reduced. Preventing neuronal death with tetrodotoxin or 6-cyano-7-nitroquinoxaline-2,3-dione rescued blood vessels, suggesting that vessel loss is not due to OGD per se but a consequence of neuronal death. Induction of excitotoxic neuronal death with AMPA caused widespread neurodegeneration, but vessel reduction was confined to CA1. In dentate gyrus without neuronal loss, vessel density increased. We propose that neuronal stress and death influence maintenance, loss and remodeling of the neurovasculature and that the type of vascular response is in addition determined by local factors within the hippocampus.

Determinants of plasma copeptin: A systematic investigation in a pediatric mechanical ventilation model

Copeptin, the C-terminal part of the arginine vasopressin precursor peptide, holds promise as a diagnostic and prognostic plasma biomarker in various acute clinical conditions. Factors influencing copeptin response in the critical care setting are only partially established and have not been investigated systematically. Using an in vivo infant ventilation model (Wistar rats, 14 days old), we studied the influence of commonly occurring stressors in critically ill children. In unstressed ventilated rats basal median copeptin concentration was 22pmol/L. In response to respiratory alkalosis copeptin increased 5-fold, while exposure to hypoxemia, high PEEP, hemorrhage, and psycho-emotional stress produced a more than 10-fold increase. Additionally, we did not find a direct association between copeptin and acidosis, hypercapnia, and hyperthermia. Clinicians working in the acute critical care setting should be aware of factors influencing copeptin plasma concentrations. Moreover, our results do have implications for animal studies in the field of stress research.

Focal necrosis and disturbed myelination in the white matter of newborn infants: a tale of too much or too little oxygen.

White matter disease in preterm infants comes along with focal destructions or with diffuse myelination disturbance. Recent experimental work with transgenic mice paves the way for a unifying molecular model for both types of brain injury, placing oxygen sensing by oligodendrocyte precursor cells (OPCs) at the center stage. Mice genetically altered to mimic high local oxygen tension in oligodendroglia lineage cells (via deletion of hypoxia-inducible factor, HIF) develop white matter disease resembling cystic periventricular leukomalacia within the first 7 days of life. Mice in which local hypoxia is mimicked in oligodendroglial cells (via genetic inhibition of HIF decay) display arrested OPC maturation and subsequent hypomyelination, reminiscent of the diffuse white matter disease observed in preterm infants and infants with congenital heart disease. These recent experimental findings on oxygen sensing and myelination are awaiting integration into a clinical framework. Gene regulation in response to hyperoxia or hypoxia, rather than oxidative stress, may be an important mechanism underlying neonatal white matter disease.

Salivary and hair glucocorticoids and sleep in very preterm children during school age

Very preterm birth involves increased stress for the child, which may lead to programming of the hypothalamic-pituitary-adrenal (HPA) axis activity and poor sleep in later life. Moreover, there is evidence for a relationship between HPA axis activity and sleep. However, research with objective sleep measures in very preterm children during school-age is rare. Eighty-five healthy children born very preterm (<32nd gestational week) and 91 full-term children aged 7-12 years were recruited for the present study. To assess HPA axis activity, salivary cortisol was measured at awakening, 10, 20, and 30min later. In addition, hair cortisol and cortisone concentrations were quantified using liquid chromatography tandem mass spectrometry to assess cumulative endocrine activity over the preceding months. One night of in-home polysomnographic sleep assessment was conducted to assess sleep duration, sleep continuity, and sleep architecture. Children born very preterm showed significantly lower levels of cortisol at awakening and lower overall post-awakening cortisol secretion, lower cortisone in hair, and earlier sleep onset than full-term children. Across the whole sample, overall post-awakening cortisol secretion was positively related to sleep onset time and negatively to sleep duration. The association between prematurity status and post-awakening cortisol secretion was partially mediated by earlier sleep onset time. In conclusion, this study provides evidence for a possible down-regulation of the HPA axis activity and slightly earlier sleep phase in very preterm children during school age.

Down-regulation of the inhibitor of growth 1 (ING1) tumor suppressor sensitizes p53-deficient glioblastoma cells to cisplatin-induced cell death

Impaired tumor suppressor functions, such as deficient p53, are characteristic for glioblastoma multiforme (GBM) and can cause resistance to DNA-damaging agents like cisplatin. We have recently shown that the INhibitor of Growth 1 (ING1) tumor suppressor is down-regulated in malignant gliomas and that the decrease of ING1 expression correlates with histological grade of malignancy, suggesting a role for ING1 in the pathogenesis and progression of malignant gliomas. Based on this background, the purpose of our current study was to examine the potential impact of ING1 protein levels on DNA-damage response in GBM. Using LN229 GBM cells, which express ING1 proteins and harbor mutant TP53, we are the first to show that DNA damage by cisplatin or ionizing radiation differentially induced the two major ING1 splicing isoforms. The p47ING1a isoform, that promotes deacetylation of histones, thus formation of heterochromatic regions of DNA, which are less susceptible to DNA damage, was preferentially induced by >50-fold. This might represent a response to protect DNA from damage. Also, ING1 knockdown by siRNA accelerated transit of cells through G1 phase, consistent with ING1 serving a tumor suppressor function, and caused cells to enter apoptosis more rapidly in response to cisplatin. Our results indicate that malignant gliomas may down-regulate ING1 to allow more efficient tumor growth and progression. Also, ING1 down-regulation may sensitize GBM cells with deficient p53 to treatment with cisplatin.

Who Plays the Strings in Newborn Analgesia at Birth, Vasopressin or Oxytocin?

For many years, oxytocin has been viewed as the primary hormone edging endocrinology, behavior, and pain in mothers and infants around parturition. Very recent work puts the vasopressin receptor 1A in the focus of peripheral analgesia and pain relief in respect to circulating vasopressin and oxytocin. Here, we present a concise overview on these new findings, will discuss them in context of parturition, and will outline new avenues.

Copeptin: a marker for stress reaction in fetuses with intrauterine growth restriction

OBJECTIVE To compare venous cord plasma concentrations of 4 vasoactive peptide precursors: carboxy-terminal proarginine vasopressin, CT-prondothelin (ET)-1, midregional proadrenomedullin, and MR-proatrial natriuretic peptide, between fetuses with intrauterine growth restriction and appropriate for gestational age controls. STUDY DESIGN Matched-pair analysis of 12 fetuses with significant intrauterine growth restriction and 42 healthy appropriate for gestational age control fetuses. All infants were singletons, delivered by elective section after 34 weeks and without chromosomal abnormalities. RESULTS Umbilical cord plasma copeptin levels (median [range]) were 4-fold higher in intrauterine growth restriction infants than in matched appropriate for gestational age controls: 23.2 (6.7-449) vs 5.1 (2.5-53) pmol/L (P < .001). Multivariate regression analysis revealed an association between copeptin and umbilical artery resistance index z-score (P = .034). The 3 other precursor peptides showed no changes. CONCLUSION High copeptin concentrations in the cord blood of intrauterine growth restriction newborns reflect a fetal stress response and support the fetal programming hypothesis.

Erythropoietin and ischemic conditioning – why two good things may be bad

The production of red blood cells is under the tight control of erythropoietin (Epo), and administration of recombinant human Epo (rhEpo) and its derivatives has become a well-established way to treat and prevent anaemia in various patients, including preterm newborn infants (1). Beyond treating anaemia, significant interest in Epo, as a novel cytoprotective agent in both neuronal and vascular systems, has been fuelled by the discovery that Epo and its receptor play a significant biological role in tissues outside the haematopoietic system. Epo has been identified as a neuroprotective agent in a wide variety of experimental models, from neuronal cell culture to in vivo brain injury. These studies suggest that Epo may protect cells against damage caused by such diverse pathological conditions as stroke, cortical trauma, inflammation, demyelinating conditions, radiation, cytostatic agents, toxin-induced epileptic seizures and retinal ischemia. Despite the widespread use of rhEpo for treatment and prevention of anaemia (rhEpo actually constitutes the fourth top-selling therapeutic category behind statins, protonpump inhibitors and antipsychotics (2)) and the wealth of experimental data demonstrating the potent neuroprotective properties of Epo, astonishingly few clinical observations demonstrate any direct effect of Epo in human neurological diseases. In very low birth weight preterm infants, administration of rhEpo in a variety of dosages up to 2100 U/kg/week intravenously has been tested in several randomized controlled trials aimed at the prevention of anaemia. As expected, rhEpo significantly reduced the total volume of blood transfused and the number of transfusions per infant, but had no effect on neurodevelopmental outcome and increased the rate of severe retinopathy of prematurity (1). In adults, to date only one randomized controlled clinical trial has shown some neuroprotective effects of rhEpo in adults suffering from stroke (3). De-

Plasma Levels of Macrophage Migration Inhibitory Factor and d-Dopachrome Tautomerase Show a Highly Specific Profile in Early Life.

Macrophage migration inhibitory factor (MIF) is a pleiotropic, constitutively expressed, pro-inflammatory cytokine and an important regulator of immune responses. d-dopachrome tautomerase (DDT), a newly described member of the MIF protein superfamily, shares sequence homology and biological activities with MIF. We recently reported that high expression levels of MIF sustain innate immune responses in newborns. Here, we elected to further characterize age-dependent MIF expression and to define whether DDT shares a similar expression profile with MIF. Therefore, we delineated the circulating concentrations of MIF and DDT throughout life using a large cohort of 307 subjects including fetuses, newborns, infants, children, and adults. Compared to levels measured in healthy adults (median: 5.7 ng/ml for MIF and 16.8 ng/ml for DDT), MIF and DDT plasma concentrations were higher in fetuses (median: 48.9 and 29.6 ng/ml), increased further at birth (median: 82.6 and 52.0 ng/ml), reached strikingly elevated levels on postnatal day 4 (median: 109.5 and 121.6 ng/ml), and decreased to adult levels during the first months of life. A strong correlation was observed between MIF and DDT concentrations in all age groups (R = 0.91, P < 0.0001). MIF and DDT levels correlated with concentrations of vascular endothelial growth factor, a protein upregulated under low oxygen tension and implicated in vascular and lung development (R = 0.70, P < 0.0001 for MIF and R = 0.65, P < 0.0001 for DDT). In very preterm infants, lower levels of MIF and DDT on postnatal day 6 were associated with an increased risk of developing bronchopulmonary dysplasia and late-onset neonatal sepsis. Thus, MIF and DDT plasma levels show a highly specific developmental profile in early life, supporting an important role for these cytokines during the neonatal period.