Date of Award
2020
Document Type
Dissertation
Degree Name
Ph.D.
Organizational Unit
College of Natual Science and Mathematics, Biological Sciences
First Advisor
David Patterson
Second Advisor
Daniel Linseman
Third Advisor
Gareth Eaton
Fourth Advisor
Joseph K. Angleson
Fifth Advisor
Dinah Loerke
Sixth Advisor
Ali Azadani
Keywords
De novo purine synthesis, Metabolomics, Purine, Transcriptomics
Abstract
Purines are a class of nitrogenous bases and are essential small molecules to life. Purines are used within the cell as genetic information carriers, energy currency, signaling molecules, and cofactors for multiple processes. They are formed through de novo and salvage pathways found in cells across the phylogenetic tree. The substrates of enzymes within de novo purine synthesis are known to influence other processes within the cell, such as energy homeostasis. In humans, de novo purine synthesis disorders are rare, with around 100 people identified. These patients exhibit a range of phenotypes, with varying degrees of mental retardation, seizure activity, facial and body dysmorphic features, autistic features, respiratory failure, and congenital blindness. To date, the explanation of phenotypes associated with these disorders remains elusive and as such, no effective therapeutic has been identified. Rare disorders are often caused by a single genetic mutation and studying rare disorders can providing key insight into processes regulated by that specific enzyme. In this body of work, I use transcriptomic profiling techniques to provide cellular and organismal process characterization of a novel cellular model of de novo purine deficiency in three CRISPR generated HeLa cell lines. I examine the de novopurine synthesis enzymes ADSL, GART, and ATIC. Processes identified influenced by de novopurine dysregulation identified are focused around neural, embryonic, organ, and placental development, epithelial to mesenchymal transition, fatty acid and inflammatory response, muscle function, tumorigenesis, oxidative stress responses, as well as TGFβ/SMAD signaling among others. Metabolomic profiling was employed to bolster transcriptomic findings, with aberrations of metabolic pathways involved in energy production, vitamin B6 and B5 metabolism, oxidative stress responses, lipids, amino acids, among others. My findings highlight areas in which de novo purine synthesis enzymes influence cellular processes responsible for cellular and organismal function and represent novel avenues of continued research.
Publication Statement
Copyright is held by the author. User is responsible for all copyright compliance.
Rights Holder
Randall Craig Mazzarino
Provenance
Received from ProQuest
File Format
application/pdf
Language
en
File Size
237 p.
Recommended Citation
Mazzarino, Randall Craig, "Cellular and Organismal Ramifications of de novo Purine Synthesis Dysregulation" (2020). Electronic Theses and Dissertations. 1807.
https://digitalcommons.du.edu/etd/1807
Copyright date
2020
Discipline
Cellular biology, Biology, Biophysics