Date of Award
5-2024
Degree Type
Thesis
Degree Name
Master of Science
Department
Biology
Program
Integrated Biosciences (MS)
First Advisor/Chairperson
Dr. Matthew Jennings
Abstract
Isocitrate dehydrogenase 1 (IDH1) is a biomarker in gliomas which provides valuable information on patient prognosis and treatment. Targeted therapies including maximal surgical resection and pharmaceutical mutant IDH inhibitors offer life extending benefits; however, currently implemented assays for the detection of IDH1 mutations do not function in a timely manner and cannot offer convenient monitoring of mutational status during treatment. Here, we describe the development and characterization of a novel dual diagnostic system consisting of a fluorescent biosensor and genotyping panel, which may collectively allow for the preoperative, intraoperative, and/or postoperative elucidation of IDH1 mutational status. The fluorescent biosensor quantifies D-2-Hydroxyglutarate (D-2-HG), an oncometabolite and surrogate marker of IDH1 mutations, while the genotyping panel identifies the specific IDH1-R132 single nucleotide variant (SNV) present. The biosensor’s robust function in biological fluids suggests its powerful utility in liquid biopsies and monitoring remission vs. recurrence, while the genotyping panel aids in the genetic detection of oncogenic missense mutations. In our hands, both assays have demonstrated 100% accuracy in discriminating between IDH1 wildtype and IDH1 mutant glioma tumors, offering exciting preliminary findings that support the implementation of these rapid diagnostic tools. In addition, the fluorescent biosensor detects depletion of D-2-HG resulting from mutant IDH inhibitors, suggesting its value in the future development of novel pharmaceutical treatments targeting mutant IDH.
Recommended Citation
Choate, Kristian A., "Förster Resonance Energy Transfer-Based Quantification of D-2-Hydroxyglutarate & Locked Nucleic Acid Loop-Mediated Isothermal Amplification Mediated Detection of IDH1-R132 Single Nucleotide Variants in Glioma" (2024). All NMU Master's Theses. 837.
https://commons.nmu.edu/theses/837
Access Type
Open Access
Justification for Restricting Access
We are currently in the process of publishing our work with FRET in addition to obtaining patents for LNA-LAMP and D-2-HG FRET.