Date on Master's Thesis/Doctoral Dissertation

5-2022

Document Type

Doctoral Dissertation

Degree Name

Ph. D.

Department

Biology

Degree Program

Biology, PhD

Committee Chair

Menze, Michael

Committee Co-Chair (if applicable)

Perlin, Michael

Committee Member

Perlin, Michael

Committee Member

Schultz, David

Committee Member

Konkle, Mary

Committee Member

Geldenhuys, Werner

Author's Keywords

Mitochondria; MitoNEET; Galactose; Thiazolidinediones; HepG2

Abstract

This dissertation explores the relationship between mitochondrial physiology and development of therapeutics. Mitochondrial dysfunction is associated with both acute and chronic forms of pathophysiology. This work aims to address development efforts at the cell culture and drug-target levels with respect to mitochondria. At the cell culture level, I characterize an approach that has been shown to improve the physiological dependency on mitochondria in tumor-derived cells. I demonstrate that prolonged replacement of glucose with galactose in culture medium induces a global metabolic shift in hepatocellular carcinoma (HepG2) cells to closer reflect a primary hepatocyte phenotype (Chapter 2). I characterize this shift by performing metabolomics and transcriptomics on HepG2 cells adapted to either glucose or galactose-based media for several weeks. At the drug development level, I explore the relationship between the thiazolidinedione (TZD) class of drugs and mitochondrial physiology. I characterize the effects of a classical TZD (pioglitazone) and two experimental TZDs (NL-1 and NL-2) on mitochondrial bioenergetics in cells derived from mammalian liver, muscle, and adipose tissues (Chapter 3). At the target development level, I examine biochemical activities that may regulate the function of a mitochondrial [2Fe-2S] protein which has been shown to bind TZDs. I provide evidence that purified MitoNEET is controlled at multiple biochemical levels by small molecular weight biological thiols and lipid-derived electrophiles (Chapter 4). Finally, I examine the role of both TZDs and NEET proteins in the regulation of cell death. I specifically demonstrate a unique role for NL-1 in protecting HepG2 cells against ferroptotic cell death (Chapter 5). Taken together, these studies provide insights that can leverage mitochondrial physiology to develop novel and effective treatments for a range of pathophysiological conditions.

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