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.
","abstract_html":"<p>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 <em>de novo</em> and salvage pathways found in cells across the phylogenetic tree. The substrates of enzymes within <em>de novo</em> purine synthesis are known to influence other processes within the cell, such as energy homeostasis. In humans, <em>de novo</em> 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 <em>de novo</em> purine deficiency in three CRISPR generated HeLa cell lines. I examine the <em>de novo</em>purine synthesis enzymes ADSL, GART, and ATIC. Processes identified influenced by <em>de novo</em>purine 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 <em>de novo</em> purine synthesis enzymes influence cellular processes responsible for cellular and organismal function and represent novel avenues of continued research.</p>","abstract_has_math":false,"creators":["Mazzarino, Randall Craig"],"institution":null,"degree_name":"Ph.D.","degree_level":"Dissertation","degree_discipline":null,"degree_department":null,"school":null,"contributors":["David Patterson","Daniel Linseman","Gareth Eaton","Joseph K. Angleson","Dinah Loerke","Ali Azadani"],"advisors":[],"committee_chairs":[],"committee_members":[],"year":2020,"date_issued":"2020-01-01T08:00:00Z","date_published":"2020-01-01T08:00:00Z","updated_at":"2026-07-24T02:02:19Z","subjects":["De novo purine synthesis","Metabolomics","Purine","Transcriptomics","Biochemistry, Biophysics, and Structural Biology","Biology","Biophysics","Cell and Developmental Biology","Cell Biology"],"languages":["en"],"rights":["Copyright is held by the author. User is responsible for all copyright compliance.
"],"rights_urls":[],"identifier_entries":[]},"links":{"outbound_url":"https://digitalcommons.du.edu/etd/1807","outbound_label":"Repository record","outbound_source":"dc:identifier"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor","label":"Contributor","values":["David Patterson","Daniel Linseman","Gareth Eaton","Joseph K. Angleson","Dinah Loerke","Ali Azadani"]},{"key":"dc:creator","label":"Author","values":["Mazzarino, Randall Craig"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date.available","label":"Dc Date Available","values":["2021-02-25T08:00:00Z"]},{"key":"thesis:degree_level","label":"Degree Level","values":["Dissertation"]},{"key":"thesis:degree_name","label":"Degree Name","values":["Ph.D."]}]},{"id":"subjects_keywords","label":"Subjects and Keywords","entries":[{"key":"dc:subject","label":"Dc Subject","values":["De novo purine synthesis","Metabolomics","Purine","Transcriptomics","Biochemistry, Biophysics, and Structural Biology","Biology","Biophysics","Cell and Developmental Biology","Cell Biology"]}]},{"id":"language_rights","label":"Language and Rights","entries":[{"key":"dc:language","label":"Dc Language","values":["en"]},{"key":"dc:rights","label":"Dc Rights","values":["Copyright is held by the author. User is responsible for all copyright compliance.
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"]},{"key":"dc:format","label":"Dc Format","values":["application/pdf"]},{"key":"dc:title","label":"Title","values":["Cellular and Organismal Ramifications of de novo Purine Synthesis Dysregulation"]}]}],"canonical_facts":{"dc:contributor":["David Patterson","Daniel Linseman","Gareth Eaton","Joseph K. Angleson","Dinah Loerke","Ali Azadani"],"dc:creator":["Mazzarino, Randall Craig"],"dc:date.available":["2021-02-25T08:00:00Z"],"dc:description.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.
"],"dc:format":["application/pdf"],"dc:identifier":["https://digitalcommons.du.edu/etd/1807"],"dc:language":["en"],"dc:rights":["Copyright is held by the author. User is responsible for all copyright compliance.
"],"dc:subject":["De novo purine synthesis","Metabolomics","Purine","Transcriptomics","Biochemistry, Biophysics, and Structural Biology","Biology","Biophysics","Cell and Developmental Biology","Cell Biology"],"dc:title":["Cellular and Organismal Ramifications of de novo Purine Synthesis Dysregulation"],"thesis:degree_level":["Dissertation"],"thesis:degree_name":["Ph.D."]},"updated_at":"2026-07-24T02:02:19Z"}