ABSTRACT
MeDALL (Mechanisms of the Development of ALLergy; EU FP7-CP-IP; Project No: 261357; 2010-2015) has proposed an innovative approach to develop early indicators for the prediction, diagnosis, prevention and targets for therapy. MeDALL has linked epidemiological, clinical and basic research using a stepwise, large-scale and integrative approach: MeDALL data of precisely phenotyped children followed in 14 birth cohorts spread across Europe were combined with systems biology (omics, IgE measurement using microarrays) and environmental data. Multimorbidity in the same child is more common than expected by chance alone, suggesting that these diseases share causal mechanisms irrespective of IgE sensitization. IgE sensitization should be considered differently in monosensitized and polysensitized individuals. Allergic multimorbidities and IgE polysensitization are often associated with the persistence or severity of allergic diseases. Environmental exposures are relevant for the development of allergy-related diseases. To complement the population-based studies in children, MeDALL included mechanistic experimental animal studies and in vitro studies in humans. The integration of multimorbidities and polysensitization has resulted in a new classification framework of allergic diseases that could help to improve the understanding of genetic and epigenetic mechanisms of allergy as well as to better manage allergic diseases. Ethics and gender were considered. MeDALL has deployed translational activities within the EU agenda.
Subject(s)
Hypersensitivity/diagnosis , Hypersensitivity/therapy , Precision Medicine/methods , Systems Biology/methods , Disease Management , European Union , Health Policy , Humans , Hypersensitivity/etiology , Hypersensitivity/prevention & control , Immunization , Immunoglobulin E/immunology , Inventions , Prognosis , World Health OrganizationABSTRACT
Whey Acidic Protein (WAP) gene expression is restricted to the pregnant and lactating mammary gland. We have recently defined a negative regulatory element (NRE) in the WAP promoter which interacts with a factor (NBF) present in all nonWAP expressing cells (Kolb et al., 1994; J. Cell. Biochem. 56:245-261). Here we characterise this factor and show that although it is not related to a number of known transcription factors, including AP-1, NF-1 and SP-1, it may also be involved in controlling the expression from the mouse mammary tumour virus promoter. Three proteins that bind to the WAP-NRE have been identified, one of which is a 53kDa nuclear protein. This protein is present in nonWAP expressing cells, suggesting that it is responsible for limiting WAP expression to the pregnant and lactating mammary gland. This protein has been partially purified and its binding to the WAP-NRE is not appreciably affected by high salt concentrations.
Subject(s)
DNA-Binding Proteins/metabolism , Mammary Glands, Animal/metabolism , Milk Proteins/genetics , Nuclear Proteins/metabolism , Promoter Regions, Genetic , Repressor Proteins/metabolism , Animals , Cell Nucleus/metabolism , DNA/metabolism , Female , Gene Expression Regulation , Mice , Molecular Weight , Pregnancy , Protein Binding , RNA, Messenger/geneticsABSTRACT
Expression of the whey acidic protein (WAP) gene is tightly regulated in a tissue and developmental stage specific manner, in that the WAP gene is exclusively expressed in the mammary gland during pregnancy and lactation. Using both deletion and competition analyses, evidence is provided for the existence of a negative regulatory element (NRE) in the WAP promoter located between -413 and -93 with respect to the WAP transcriptional initiation site. This NRE dramatically decreases transcription from linked heterologous promoter-reporter gene constructs. The activity of NRE requires WAP promoter sequences that are 230 bp apart since subfragments of the NRE fail to inhibit transcription of adjoining reporter genes. Nuclear extracts from different cell types, in which the WAP gene is not active, contain a protein or complex that specifically interacts with the entire NRE but not with subfragments of it. The contact points between this protein (NRE binding factor [NBF]) and the NRE element have been partially determined. Mutation of the implicated nucleotides severely reduces the ability of NBF to bind, and such mutated promoter fragments fail to alleviate transcriptional repression in competition experiments. This suggests that NBF binding to the NRE is at least in part responsible for the negative regulation of the WAP promoter. Since NBF is not detectable in the lactating mammary gland, where the WAP gene is expressed, we speculate that it may be a determinant of the expression spectrum of the WAP gene.
