P450 Induction in Cryopreserved Hepatocytes from PXR and CAR Nuclear Receptor Knock-out Rats
Poster Jun 19, 2018
Kevin P. Forbes1, Kirsten Amaral2 and Albert P. Li2
Here, we report the isolation and preliminary characterization of cryopreserved hepatocytes from male PXR and CAR knockout (SD) rats and PXR/CAR double knockout (SD) rats (Horizon Discovery - SAGE Labs). We were successful in the isolation and cryopreservation of the hepatocytes from the three knockout models, yielding hepatocytes with high (>80%) viability and plateability. The cryopreserved hepatocytes from wildtype and knockout rats were cultured for the evaluation of gene expression in the presence and absence of PXR and CAR ligands. The cryopreserved hepatocytes were recovered using UCRM (IVAL Inc.) and plated in 24-well collagen-coated plates. The hepatocytes formed near confluent monolayer cultures with epithelial morphology typical of primary cultured rat hepatocytes. After culturing overnight, medium was changed to protein free induction medium for rat hepatocytes (RHIM, IVAL Inc.). The hepatocytes were treated the next day with the PXR-ligand pregnenlone-16α-carbonitrile (PCN) and the CAR-ligand 3,3ʹ,5,5ʹ-tetrachloro-1,4-bis(pyridyloxy) benzene (TCPOBOP). Our results show that PXR was required for PCN-activation of Cyp2b2, Cyp3a23/3a1, Cyp3a18, Cyp2c11 and Slco1a2 (Oatp1a2) gene expression, and that CAR was required for TCPOBOP-activation of Cyp2b2 and Cyp3a23/3a1 gene expression, in good agreement with data obtained from in vivo rat knockout models1, human primary hepatocytes, nuclear receptor knock-out cell lines and mouse knock-out models. Hepatocytes from the PXR and CAR knockout rats therefore represent a useful ex vivo complement to the in vivo models and tools for drug development, especially for functional studies on metabolic pathways involving nuclear receptors.
Characterization of a Type 2 diabetes-associated islet-specific enhancer cluster in STARD10 by genome editing of EndoC-βH1 cellsPoster
Genome-wide association studies (GWAS) have identified more than 100 genetic loci associated with type 2 diabetes. The majority of these are located in the intergenic or intragenic regions suggesting that the implicated variants may alter chromatin conformation. This, in turn, is likely to influence the expression of nearby or more remotely located genes to alter beta cell function. At present, however, detailed molecular and functional analyses are still lacking for most of these variants. We recently analysed one of these loci and mapped five causal variants in an islet-specific enhancer cluster within the STARD10 gene locus. Here, we aimed to understand how these causal variants influence b-cell function by alteration of the chromatin structure of enhancer clusterREAD MORE
Psychiatric Risk Gene Cacna1c and Early Life Stress: Potential Gene-Environment interactions?Poster
Early life stress (ELS) is highly associated with development of psychopathology
and mood disorders in adulthood. Genetic studies have identified variation in the gene calcium voltage-gated channel subunit alpha1C (CACNA1C) to increase risk for several psychiatric disorders. This poster assessed the expression of Cacna1c following prepubertal stress.
Treatment Options for Chronic Parvovirus Viremia in Pediatric Heart Transplant Patients in a Tertiary Care CenterPoster
This abstract discusses three cases of pediatric heart transplant patients who suffered from parvovirus (B19) infection. Of these patients, two ( B & C) responded well to standard intravenous Ig therapy. Patient A however, did not respond to standard treatment and was begun on subcutaneous Ig, which effectively diminished his viral load. Thus, subcutaneous Ig infusions might serve as a second line treatment for transplant patients with parvovirus who do not respond well to the standard approach.READ MORE
3rd International Conference on Diagnostic Microbiology and Infectious Diseases
Sep 24 - Sep 25, 2018
5th edition of the International Conference Clinical Oncology and Molecular Diagnostics
Jun 17 - Jun 18, 2019