Date of Completion

2021

Thesis Type

College of Arts and Science Honors

Department

Biomedical and Health Sciences

First Advisor

Paula Deming, Ph.D.

Second Advisor

Matthias Brewer, Ph.D.

Keywords

STK11, Lung Adenocarcinoma, Variants of Unknown Significance, Next Generation Sequencing

Abstract

Lung carcinomas are the leading cause of cancer-related mortality in Vermont, and approximately 30% of non-small cell lung carcinomas (NSCLC) have mutations in the tumor suppressor gene Serine/Threonine Kinase 11 (STK11). Acting as a cellular energy sensor, STK11 regulates cell growth, metabolism, and migration. In KRAS-driven NSCLC, STK11 loss of function mutations correlate with more aggressive lung tumors, metastasis, and poorer response to immunotherapy. STK11 functional status is therefore clinically useful to predict disease prognosis and guide treatment. Mutations in STK11 are functionally characterized as (likely) pathogenic, (likely) benign or a variant of unknown significance (VUS). Clinicians at UVMMC utilized next generation sequencing of patient lung adenocarcinomas to identify genetic mutations in genes known to impact prognosis or treatment, including STK11. Twenty-eight missense alterations in the coding region of the STK11 gene were identified as un- and under-characterized variants. This research presents data that characterizes the function of the twenty-eight STK11 missense VUS using a tissue culture reconstitution model and p53-dependent luciferase reporter assay. When our data was compared to twenty-two different in silico predictive algorithms, there were inconsistent functional predictions for the majority of the variants, which highlights the importance of creating a reliable functional assay that can rapidly asses STK11 missense variants. Additionally, we developed an in vitro radioactive kinase assay to analyze the biochemical kinetics of WT-STK11 and the twenty-eight variants. Our results provide information that can guide clinical utilization of immunotherapy and help predict prognosis for patients with NSCLC.

Available for download on Sunday, May 15, 2022

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