Event Title

Session 3E: LKB1 in Pathogenesis of Multiple Sclerosis and Experimental Autoimmune Encephalomyelitis (Mouse Model of MS)

Session Number

Session 3E: 2nd Presentation

Advisor(s)

Douglas Feinstein, University of Illinois at Chicago

Location

Room A113

Start Date

28-4-2017 1:15 PM

End Date

28-4-2017 2:30 PM

Abstract

Multiple Sclerosis (MS) is the most common neurodegenerative disease that targets people from 10-80 years of age. Our lab previously showed that a mutation (variant) in the gene STK11 increased the risk of getting MS. However, the mechanisms of action of the SKT11 gene and its role in MS is not clearly understood. We demonstrate here that knockout (KO) of STK11 from cerebral cortex microglial cells (SIM-A9) increases the inflammatory responses of the cells. To do this, cultures of SIM-A9 cells were treated with targeting and non-targeting siRNA (silencing RNA) for one day to KO the STK11 mRNA from the cells. The cells were then treated with LPS (lipopolysaccharide, a strong stimulator) and TII (a combination of cytokines). One day later we prepared mRNA from the cells and then we used quantitative PCR to observe differences between the STK11 mRNA levels in KO and control cells. Analysis of nitrite production (an inflammatory marker) in cells with STK11 present versus KO identified an increase in inflammatory response in the KO cells after LPS + TII treatment. In the second part of the study, a similar procedure was used on primary microglia, which gave similar results. qPCR was also used to test if other mRNAs are affected by the STK11 KO, and showed that some mRNA were also increased, including the mRNA that codes for NOS2, the enzyme that makes nitrites. Collectively, these data indicate that KO of STK11 results in an upregulation of inflammatory responses in the SIM-A9 microglial cell line and in primary microglial cells, suggesting that mutations in STK11 which lower STK11 mRNA levels could lead to worse inflammation in MS patients.

Comments

Additional team members: Dr. Sergey Kalinin

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Apr 28th, 1:15 PM Apr 28th, 2:30 PM

Session 3E: LKB1 in Pathogenesis of Multiple Sclerosis and Experimental Autoimmune Encephalomyelitis (Mouse Model of MS)

Room A113

Multiple Sclerosis (MS) is the most common neurodegenerative disease that targets people from 10-80 years of age. Our lab previously showed that a mutation (variant) in the gene STK11 increased the risk of getting MS. However, the mechanisms of action of the SKT11 gene and its role in MS is not clearly understood. We demonstrate here that knockout (KO) of STK11 from cerebral cortex microglial cells (SIM-A9) increases the inflammatory responses of the cells. To do this, cultures of SIM-A9 cells were treated with targeting and non-targeting siRNA (silencing RNA) for one day to KO the STK11 mRNA from the cells. The cells were then treated with LPS (lipopolysaccharide, a strong stimulator) and TII (a combination of cytokines). One day later we prepared mRNA from the cells and then we used quantitative PCR to observe differences between the STK11 mRNA levels in KO and control cells. Analysis of nitrite production (an inflammatory marker) in cells with STK11 present versus KO identified an increase in inflammatory response in the KO cells after LPS + TII treatment. In the second part of the study, a similar procedure was used on primary microglia, which gave similar results. qPCR was also used to test if other mRNAs are affected by the STK11 KO, and showed that some mRNA were also increased, including the mRNA that codes for NOS2, the enzyme that makes nitrites. Collectively, these data indicate that KO of STK11 results in an upregulation of inflammatory responses in the SIM-A9 microglial cell line and in primary microglial cells, suggesting that mutations in STK11 which lower STK11 mRNA levels could lead to worse inflammation in MS patients.