Graduation Date

5-2014

Document Type

Master's Thesis

Degree Name

Master of Science

Department or Program

Biological Sciences

Department or Program Chair

Kiowa Bower, Ph.D.

First Reader

Arvind Ramanathan, PhD

Second Reader

Dorn Carranza, PhD

Abstract

Naturally occurring small molecules (metabolites, signaling intermediates) are a critical component of the information flow in biology, along with DNA, RNA, and proteins. Metabolomics is an analytical approach that seeks to comprehensively analyze naturally occurring small molecules and quantify their dynamic changes in biological systems. In recent years metabolomics has begun to provide understanding of the metabolic basis of different diseases, such as heart disease, cancer, and diabetes. Our lab built a High-Performance Liquid-Chromatography Mass-Spectrometry (HPLC-MS) based metabolomics platform to analyze metabolites from mammalian cells, spent cellular media, and model organisms such as C. elegans. We used C. elegans to elucidate the metabolic changes seen after treatment with Metformin, which is a known activator of the AMPK pathway. Cancer cells exhibit high levels of glycolysis producing large amounts of lactate; circumventing the mitochondrial pathway. This phenomenon is known as the Warburg effect. We hypothesize that cancer metabolism is epigenetically regulated. Epigenetics refers to inheritable traits that are not due to alterations in the primary DNA sequence. DNA methylation is an important epigenetic modification. DNA methylation mainly occurs in the CpG islands of the promoter region of genes. It is believed that during cancer development de novo DNA methyltransferases methylate tumor-suppressing genes allowing cancer cells to proliferate uninhibited. There are two de novo DNMTs, DNMT3A, and DNMT3B. These DNMTs establish the pattern of methylation. We examined de novo DNMT-mediated control of cellular metabolism, identifying global changes in metabolism, as well as differential sensitivity towards glycolytic and mitochondrial inhibitors.

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