A High-Throughput Approach to Characterizing ARV1 on the Regulation of Lipid Homeostasis Uncovers a Novel Interaction with Epidermal Growth Factor Receptor

Author(s)

Nicholas Anthony Wachowski, Rowan University

Date Approved

4-2021

Document Type

Thesis

Degree Name

Master of Science in Molecular Pathology and Immunology

Department

Molecular Biology

College

Graduate School of Biomedical Sciences

Sponsor

Genesis Biotechnology Group, Oncoveda, Institute of Metabolic Disorders

First Advisor

Salvatore Caradonna, PhD

Committee Member 1

Scott Gygax, PhD

Committee Member 2

Joseph Nickels, PhD

Subject(s)

Sterol O-Acyltransferase, Non-alcoholic Fatty Liver Disease, Fatty Acids, Hep G2 Cells, Phosphorylation, ErbB Receptors, Transcriptome, Coenzyme A

Disciplines

Cancer Biology | Cell Biology | Immunopathology | Laboratory and Basic Science Research | Medical Cell Biology | Medical Immunology | Medical Molecular Biology | Medicine and Health Sciences | Molecular Biology | Molecular Genetics | Nutritional and Metabolic Diseases | Pathological Conditions, Signs and Symptoms

Abstract

Acyl-CoA cholesterol acyl transferase related enzyme-2 required for viability 1 (ARV1) was first recognized in Saccharomyces cerevisiae in a study done in 2000 by Tinkelenberg et al. In yeast, the deletion of ARV1 results in numerous defects including abnormal sterol trafficking [1], the reduction of sphingolipid metabolism [2], synthesis of glycosylphosphatidylinositol (GPI) anchor [3], ER stress [4], and hypersensitivity of fatty acids leading to lipoapoptosis [5]. Arv1 germline deletion in mice displayed a lean phenotype with increased energy [6]. In humans, ARV1 mutations lead to epileptic encephalopathy [7].

Non-alcoholic fatty liver disease (NAFLD) consists of simple steatosis to non-alcoholic steatohepatitis (NASH), fibrosis, cirrhosis, and hepatocellular carcinoma [8]. NAFLD is the most common liver disease worldwide affecting 25% of the global population and 33% of the population in the United States [8]. NAFLD is on the rise due to irresponsible dietary and sedentary lifestyles. Currently, there is not a pharmacological treatment for NAFLD.

The specific aims include the generation of ARV1 over expression and ARV1 knockout in HepG2 cells, to investigate gene expression profiles in each respective ARV1 cell line using Qiagen RT2 Profiler arrays and to identify novel pathways to validate Arv1 function in HepG2 cells. We found ARV1 over expression cells to be lipotoxic in HepG2 cells. We showcased EGFR phosphorylation of Arv1. We observed two different EGFR inhibitors where both “rescued” toxicity in Arv1 over expression cells.

Recommended Citation

Wachowski, Nicholas Anthony, "A High-Throughput Approach to Characterizing ARV1 on the Regulation of Lipid Homeostasis Uncovers a Novel Interaction with Epidermal Growth Factor Receptor" (2021). Graduate School of Biomedical Sciences Theses and Dissertations. 30.
https://rdw.rowan.edu/gsbs_etd/30