Sepsis remains the health-care problem associated with a high mortality despite advances in drug treatment. There is considerable interest in genetic differences between patients, which may modulate the risk of severe sepsis development and outcome. A number of genetic associations have been found in genes involved in the immune and inflammatory pathways. However it remains unclear if the disease-associated genetic polymorphisms are themselves functionally important or serving as a marker for co-inherited regulatory variants on the same haplotype. To identify genetic variation modulating gene expression present on sepsis-associated haplotypes, we propose to study candidate disease-associated genes in a model system and in patients.
A cell line model will be used comprising a panel of immortalised human B cells established from different individuals with defined genetic make-up. This will allow us to use a set of haplotypes for expression studies containing disease-associated polymorphisms. A number of stimuli will be used to resolve if haplotype-specific differences in gene expression are present in cell lines. Then candidate functional haplotypes in primary human cells from patients with severe sepsis will be investigated. For disease-associated haplotypes showing differences in expression in both the cell lines and on primary cell analysis we will investigate the underlying functional mechanisms using assays of transcription such as differential protein-DNA binding in vitro and in vivo.
This proposal will advance our understanding of how genetic differences between patients can be used to improve outcome of sepsis. It is timely due to the wealth of genetic data becoming available to resolve how genetic variation is co-inherited and will take advantage of state-of -the-art experimental approaches available in the proposed Host Centre to allow resolution of functional effects of variation on gene expression.
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