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Title:Small molecule inhibitors of lactate dehydrogenase A as an anticancer strategy
Author(s):Calvaresi, Emilia
Director of Research:Hergenrother, Paul J.
Doctoral Committee Chair(s):Hergenrother, Paul J.
Doctoral Committee Member(s):Morrissey, Jim; Shapiro, David J.; Gennis, Bob
Department / Program:Biochemistry
Discipline:Biochemistry
Degree Granting Institution:University of Illinois at Urbana-Champaign
Degree:Ph.D.
Genre:Dissertation
Subject(s):Warburg effect
cancer biology
biochemistry
lactate dehydrogenase
lactate dehydrogenase (LDH)
lactate dehydrogenase A (LDH-A)
cancer metabolism
cancer
glucose transporter-1 (GLUT-1)
glucose transporter 1 (GLUT1)
glucose transporter (GLUT)
glycoconjugation
glucose conjugation
anticancer
anticancer agents
metabolism
small molecules
Cellular Thermal Shift Assay (CETSA)
Drug Affinity Responsive Target Stability (DARTS)
Abstract:Exploiting cancer cell metabolism as an anticancer therapeutic strategy has garnered much attention in recent years. As early as the 1920s, German scientist Otto Warburg observed cancer tissues’ avid glucose consumption and high rates of aerobic glycolysis, a phenomenon now known as the Warburg effect. Today, we understand the Warburg effect is mediated by a number of complex factors, including overexpression of the insulin-independent glucose transporter GLUT-1 and overexpression of various glycolytic enzymes, including lactate dehydrogenase A (LDH-A). As the terminal enzyme of glycolysis, LDH-A catalyzes the reversible conversion of pyruvate to lactate, and in doing so, oxidizes NADH to NAD+. The lactate produced by this reaction is largely excreted into the tumor microenvironment, where it acidifies surrounding tissues and helps the tumor evade destruction by immune cells. The oxidation of NADH to NAD+ allows for continued ATP production through glycolysis by replenishing NAD+ in the absence, or reduced function, of oxidative metabolism. Cell culture and in vivo studies of LDH-A knockdown (using RNA interference) have been shown to lead to substantial decreases in cell and tumor proliferation, thus providing evidence that LDH-A would be a viable anticancer target. While various in vitro LDH-A inhibitors exist, there is a need for a potent and selective small molecule inhibitor that functions both in cells and in vivo. Here, the development and biological assessment of the N-hydroxyindole class of LDH-A inhibitors, including a series of novel dual-Warburg targeting glucose-conjugated LDH-A inhibitors, developed through a collaboration between the Hergenrother and Minutolo laboratories, is reported. The development of novel assays to assess the relative cell uptake, cell lactate production, and competition with 13C glucose for cellular entry, of NHI series compounds are also discussed. Head-to-head cellular assessments of the most promising NHI series compounds alongside literature-reported in vitro inhibitors of LDH-A are reported. Finally, efforts to directly probe the interactions of compounds with LDH-A in cell lysate and whole cells using CETSA and DARTS techniques are discussed.
Issue Date:2015-01-21
URI:http://hdl.handle.net/2142/72952
Rights Information:Copyright 2014 Emilia Calvaresi
Date Available in IDEALS:2015-01-21
Date Deposited:2014-12


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