Homo sapiens
Ontology highlight
ABSTRACT:
PROVIDER: PRJNA137073 | ENA |
REPOSITORIES: ENA
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Project description:Laron Syndrome (LS) is a rare hereditary disease in humans caused by a mutation in the growth hormone receptor gene which leads to high levels of circulating GH [1]. The disease is characterized by short statue, obesity, transient hypoglycemia during youth and low incidences of cancer and diabetes [2]. Thus making it interesting to study the underlying mechanisms. Recently a porcine model to mimic human LS was developed [3]. Growth hormone receptors are expressed on various cells of the immune system yet the extent to which dysregulation of GH might affect immune function in LS is poorly understood [4]. Therefore we decided to characterize the influence of growth hormone receptor knockout on the immune function of these animals by analyzing the proteome of their CD4+ and CD4- peripheral blood mononuclear cells (PBMC). 1. Werner, H.; Sarfstein, R.; et al. Laron Syndrome Research Paves the Way for New Insights in Oncological Investigation. Cells 2020, 9, doi:10.3390/cells9112446. 2. Werner, H.; Lapkina-Gendler, L.; et al. Genome-Wide Profiling of Laron Syndrome Patients Identifies Novel Cancer Protection Pathways. Cells 2019, 8, doi:10.3390/cells8060596. 3. Hinrichs, A.; Kessler, B.; et al. Growth hormone receptor-deficient pigs resemble the pathophysiology of human Laron syndrome and reveal altered activation of signaling cascades in the liver. Mol Metab 2018, 11, 113-128, doi:10.1016/j.molmet.2018.03.006. 4. Hattori, N. Expression, regulation and biological actions of growth hormone (GH) and ghrelin in the immune system. Growth Horm IGF Res 2009, 19, 187-197, doi:10.1016/j.ghir.2008.12.001.
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Project description:Biomarkers of responsiveness to treatment with recombinant human GH (rhGH) have only poorly been described. We aimed at identifying biomarkes of rhGH treatment in children. Our aim was to identify the genes in response to rhGH in human PBMC, that can be used as Biomarkes of growth prediction. Blood was obtained from pediatric patients before and 4 days after the growth hormone therapy. RNA was isolated from the PBMC and subsequently used to perform the whole genome cDNA microarrays. We used the type-1 microarray strategy (RNA isolated before the hormone treatment as a control, and RNA isolated after rhGH treatment). Microarrays represent PBMC RNA obtained from growth hormone treated patients (green channel = baseline, red channel = GH tretmanet for 4 days). Stanford cDNA microarrays were used. Only genes corresponding to spots with fluorescence signals of 1.5-fold over array background in both, the experimental channel (Cy5) and the reference channel (Cy3) and only genes with at least 80% interpretable data in the 9 microarray experiments were considered. Of note, the Cy5/Cy3 ratios on the arrays correspond directly to the fold up-or down-regulation of gene transcription following rhGH treatment. Subsequently, the Cy5/Cy3 ratios on arrays were log2 transformed for further analyses and the dataset was further reduced by considering only those genes with at least a 2-fold up- or down- regulation of transcription in least 3 of the 9 microarray experiments. Microarray experiments and genes were then organized by unsupervised hierarchical clustering using the Pearson correlation metric and average linkage clustering.
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