Brahmpreet Saini
Pronouns: he/him
Research Mentor(s): Elizabeth Speliotes
Co-Presenter:
Research Mentor School/College/Department: Internal Medicine / Medicine
Presentation Date: April 20
Presentation Type: Poster
Session: Session 2 – 11am – 11:50am
Room: League Ballroom
Authors: Brahmpreet Saini, Yue Chen, Elizabeth Speilotes
Presenter: 56
Abstract
Functional Genetic Studies on GCKR Variants in Human Nonalcoholic Fatty Liver Disease Background: Glucokinase regulatory protein (GKRP) is a protein that is mainly expressed in the liver and interacts with glucokinase (GCK) as means to regulate the glucose disposal and storage in both liver and pancreatic beta-cells in response to changes in glucose concentrations [1].Prior Studies have shown that GCKR also acts as a post-translational stabilizer of cellular GCK and indicates that the level of GCKR is important for regulating the level and subcellular localization of GCK [1]. However, the effect of different GCKR variants on lipid accumulation, and different expression levels on lipid accumulation, along with a molecular mechanism for GCKR regulation on GCK expression and relocalization are yet to be determined Methods and Materials: To examine the roles of GCKR in regulating the level and subcellular localization of GCK protein in different doses of glucose, Huh-7 cell lines overexpressing GCKR wildtype and mutant P446L gene and GCKR knockdown cells were used as the cellular models. These were then seeded into 24 and 96 wells in a low glucose medium (5.5 mM) and treated with varying doses of glucose (0,2, 5, and 25mM). Following an incubation period (4 – 8 hours), the cells were collected for a triglyceride assay and western blot analysis. The cells in the 96-well plate were immunofluorescence stained with anti-GCK and anti-GCKR primary antibodies. Glucose treated GCKR overexpression/knockdown HuH-7 cell lines were analyzed using high content fluorescent microscopy screening and CellProfiler software analysis. Goals and Expectations: The goal of the study is to find the effect of different GCKR variants on binding to GCK, and gain knowledge on the interaction and co-localization of GCKR, and GCK proteins in nuclei and cytoplasm of glucose treated GCKR overexpression/knockdown HuH-7 cell line. Being able to gain insight into these elements would improve our understanding of glucose and lipid physiology and the development of targeted molecular therapeutics for nonalcoholic fatty liver disease and obesity related diseases. References [1] Int. J. Mol. Sci. 2015, 16 7379
Biomedical Sciences, Interdisciplinary
