Presentation Title

Characterizing STK11 Variants of Unknown Significance Identified in Vermont Patients

Project Collaborators

David J Seward (Collaborating Mentor)

Abstract

Lung cancer accounts for 15% of newly diagnosed cancer and is the leading cause of mortality due to cancer in Vermont. Serine/threonine kinase 11 (STK11) is a tumor suppressor that is mutated in approximately 30% of patients with non-small cell lung carcinoma. Pathogenic, loss of function mutations in the STK11 gene corresponds with poorer survival and response to immunotherapy, the first line treatment for advanced-stage lung cancer patients. Clinicians use next generation sequencing (NGS) to determine the molecular patterns that accrue during tumor development and identify commonly mutated genes, allowing physicians to more effectively treat patients. In addition to known pathogenic STK11 mutations, variants of unknown significance (VUS) are frequently identified in lung adenocarcinomas, where the impact on STK11 kinase activity and function are unknown. Next generation sequencing from clinical lung adenocarcinoma samples at the University of Vermont Medical Center identified 28 VUS in the coding region of the STK11 gene that result in a single amino acid substitution. To determine whether these genetic variants impact the function of the STK11 gene, we utilized molecular cloning techniques to recreate the VUS isolated at the University of Vermont Medical Center in a mammalian expression vector for in vitro experiments. We transiently transfected these variants into an STK11 null lung adenocarcinoma cell line and utilized a p53 transcriptional assay to determine the enzymatic activity of VUS. Wild-type STK11 activates p53 and results in increased p53 driven transcription, whereas kinase dead (KD) STK11 does not induce p53 activity. Using this system, we were able to determine whether the genetic alterations in STK11 isolated from UVMMC patients result in a loss of function, which can help physicians create more personalized treatment plans for their patients.

Primary Faculty Mentor Name

Paula B Deming

Status

Undergraduate

Student College

College of Agriculture and Life Sciences

Program/Major

Microbiology

Primary Research Category

Biological Sciences

Second College (optional)

College of Nursing and Health Sciences

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Characterizing STK11 Variants of Unknown Significance Identified in Vermont Patients

Lung cancer accounts for 15% of newly diagnosed cancer and is the leading cause of mortality due to cancer in Vermont. Serine/threonine kinase 11 (STK11) is a tumor suppressor that is mutated in approximately 30% of patients with non-small cell lung carcinoma. Pathogenic, loss of function mutations in the STK11 gene corresponds with poorer survival and response to immunotherapy, the first line treatment for advanced-stage lung cancer patients. Clinicians use next generation sequencing (NGS) to determine the molecular patterns that accrue during tumor development and identify commonly mutated genes, allowing physicians to more effectively treat patients. In addition to known pathogenic STK11 mutations, variants of unknown significance (VUS) are frequently identified in lung adenocarcinomas, where the impact on STK11 kinase activity and function are unknown. Next generation sequencing from clinical lung adenocarcinoma samples at the University of Vermont Medical Center identified 28 VUS in the coding region of the STK11 gene that result in a single amino acid substitution. To determine whether these genetic variants impact the function of the STK11 gene, we utilized molecular cloning techniques to recreate the VUS isolated at the University of Vermont Medical Center in a mammalian expression vector for in vitro experiments. We transiently transfected these variants into an STK11 null lung adenocarcinoma cell line and utilized a p53 transcriptional assay to determine the enzymatic activity of VUS. Wild-type STK11 activates p53 and results in increased p53 driven transcription, whereas kinase dead (KD) STK11 does not induce p53 activity. Using this system, we were able to determine whether the genetic alterations in STK11 isolated from UVMMC patients result in a loss of function, which can help physicians create more personalized treatment plans for their patients.