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

<p>Isocitrate dehydrogenase 1 (<em>IDH1</em>) 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 <em>IDH1</em> 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 <em>IDH1 </em>mutational status. The fluorescent biosensor quantifies D-2-Hydroxyglutarate (D-2-HG), an oncometabolite and surrogate marker of <em>IDH1</em> mutations, while the genotyping panel identifies the specific <em>IDH1</em>-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 <em>IDH1</em> wildtype and <em>IDH1</em> 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.</p>

Name thesis:degree_name

Master of Science

Level thesis:degree_level

Thesis

Discipline thesis:degree_discipline

Biology

Year dc:date.available

2024

Author dc:creator

• Choate, Kristian A

Contributors dc:contributor

• Dr. Matthew Jennings

• Glioma
• IDH1
• Oncometabolite
• D-2-Hydroxyglutarate
• D-2-HG
• Loop-mediated isothermal amplification
• Peptide nucleic acids
• Förster Resonance Energy Transfer
• Locked nucleic acid
• Vorasidenib
• Diagnosis

Repository record dc:identifier

https://commons.nmu.edu/theses/837

OAI identifier oai:identifier

oai:commons.nmu.edu:theses-1823