Project description:Background: Epigenetic alteration of the genome has been shown to provide palliative effects in mouse models of certain human autoimmune disorders. We have investigated whether chromatin remodeling could provide protection against autoimmune diabetes in nonobese diabetic (NOD) mice. Treatment of female mice during the transition from prediabetic to diabetic stage (18-24 weeks of age) with the well-characterized histone deacetylase inhibitor, Trichostatin A (TSA) effectively reduced the incidence of diabetes and abrogated the ability of splenocytes to adoptively transfer the disease into immunodeficient NOD.scid mice. Protection against diabetes was accompanied by histone hyperacetylation in pancreas and spleen, increased frequency of CD4+ CD62L+ cells in the spleen, reduction in cellular infiltration of islets, restoration of normoglycemia and glucose-induced insulin release by beta cells. In vitro activation of splenic T lymphocytes derived from protected mice resulted in enhanced expression of IFN-gamma mRNA and protein without altering the expression of Il4, Il17, Il18, Inos, and Tnfa genes nor the secretion of IL-2, IL-4, IL-17 and TNF-alpha proteins. Consistently, expression of the transcription factor involved in Ifng transcription, Tbet/Tbx21 but not Gata3 and Rorgt, respectively required for the transcription of Il4 and Il17, was upregulated in activated splenocytes of protected mice. These data indicate that abrogation of autoimmune diabetes is associated with the selective upregulation of certain inducible genes in T lymphocytes. Microarray analysis was performed to determine the changes in global gene expression underlying abrogation of autoimmune diabetes by TSA-mediated epigenetic modulation and identified a distinct group of genes down-regulated during this process. Female NOD mice were treated subcutaneously with TSA (500 µg/Kg body weight) during the transition from prediabetic to diabetic stage (18-24 weeks of age), at weekly intervals. Since we sought to determine the modulation of global gene expression associated with long-term protection against diabetes, TSA treated mice were killed between 32 and 36 weeks of age and total RNA was extracted from uninduced splenocytes. RNA was also extracted from splenocytes of untreated mice that became diabetic and those that did not develop diabetes till 36 weeks of age. Blood glucose levels were determined weekly to monitor the glycemic condition in untreated and TSA-treated mice. To minimize the variability in gene expression among individual mice, RNA was extracted from splenocytes of 3 to 5 mice per group and pooled. Each sample was hybridized to microchips in duplicate. Female NOD mice were treated with Trichostatin A (500 µg/Kg body weight) between 18-24 weeks of age or left untreated.

Project description:Diabetes is a disease now affecting more than 20 million people in the United States alone. When left untreated it has numerous debilitating effects including diabetic retinopathy, which is characterized by the death of retinal neurons as well as a breakdown of the retinal vascularization. This leads to nearly 24,000 cases of blindness each year among diabetic patients. Studies have shown the importance of both nitric oxide and the gene VEGF in retinopathy, however little is known about the condition's progression. This study would determine which genes are affected early in diabetes and more generally, give insite into how neural degeneration occurs in the retina. We aim to show the gene-level changes in the diabetic mouse retina five weeks after induction of diabetes. Our laboratory's primary focus is on NO production through nNOS, iNOS and endothelial (eNOS). We hope to be able to correlate some of the transcript level changes with genes known to be involved in the NO signalling pathways. Previous data by our lab and others have shown that neuronal nitric oxide synthase (nNOS) and inducible NOS (iNOS) are upregulated under diabetic conditions in the retina. Additional work in our lab has shown that the lack of nitric oxide (NO) produced by nNOS may affect neurotransmitter production, particularly with GABA. We have also shown that mice with induced diabetes show neurochemical changes in the retina after only one to three weeks. Thus, we expect to see gene level changes associated with the retina before psysological changes in newly diabetic mice. Adult CD1 mice of random sexes are injected with the drug streptozotocin on three successive days while fasting 8 hours before the injection. This drug selectivley destroys the beta-islet cells of the pancrease. Their blood sugar levels are then tested after a resting period. Mice with glucose levels above 250mg/dL are considered diabetic from the date of the last injection. Mice are then sacrificed after the waiting period by breif isoflourine inhalation followed by decapitation. Retinas are immediately removed in cold HBSS and RNA is extracted using Trizol extraction. To get adequate amounts of RNA, we will pool four retinas (2 animals) together.

Project description:Diabetes is a disease now affecting more than 20 million people in the United States alone. When left untreated it has numerous debilitating effects including diabetic retinopathy, which is characterized by the death of retinal neurons as well as a breakdown of the retinal vascularization. This leads to nearly 24,000 cases of blindness each year among diabetic patients. Studies have shown the importance of both nitric oxide and the gene VEGF in retinopathy, however little is known about the condition's progression. This study would determine which genes are affected early in diabetes and more generally, give insite into how neural degeneration occurs in the retina. We aim to show the gene-level changes in the diabetic mouse retina five weeks after induction of diabetes. Our laboratory's primary focus is on NO production through nNOS, iNOS and endothelial (eNOS). We hope to be able to correlate some of the transcript level changes with genes known to be involved in the NO signalling pathways. Previous data by our lab and others have shown that neuronal nitric oxide synthase (nNOS) and inducible NOS (iNOS) are upregulated under diabetic conditions in the retina. Additional work in our lab has shown that the lack of nitric oxide (NO) produced by nNOS may affect neurotransmitter production, particularly with GABA. We have also shown that mice with induced diabetes show neurochemical changes in the retina after only one to three weeks. Thus, we expect to see gene level changes associated with the retina before psysological changes in newly diabetic mice. Adult CD1 mice of random sexes are injected with the drug streptozotocin on three successive days while fasting 8 hours before the injection. This drug selectivley destroys the beta-islet cells of the pancrease. Their blood sugar levels are then tested after a resting period. Mice with glucose levels above 250mg/dL are considered diabetic from the date of the last injection. Mice are then sacrificed after the waiting period by breif isoflourine inhalation followed by decapitation. Retinas are immediately removed in cold HBSS and RNA is extracted using Trizol extraction. To get adequate amounts of RNA, we will pool four retinas (2 animals) together. Keywords: dose response

			Action	DRS
	E-MTAB-5086-analysis-methods.tsv	Tabular
	E-MTAB-5086-atlasExperimentSummary.Rdata	Rdata
	E-MTAB-5086-configuration.xml	Xml
	E-MTAB-5086-percentile-ranks.tsv	Tabular
	E-MTAB-5086.condensed-sdrf.tsv	Tabular

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E-MTAB-5086

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