Date of Completion
Spring 5-1-2026
Thesis Advisor(s)
Christopher D. Heinen; Christopher Malinoski
Honors Major
Molecular and Cell Biology
Disciplines
Cancer Biology | Congenital, Hereditary, and Neonatal Diseases and Abnormalities | Medical Genetics | Medical Molecular Biology | Oncology
Abstract
Lynch syndrome is a hereditary disease caused by the inheritance of a mismatch repair gene variant. Individuals with this condition are predisposed to cancer development, most commonly colorectal cancer. Current guidelines for variant classification are based on numerous evidence categories, including functional evidence. However, many novel clinical variants are not well characterized, and evidence is difficult to obtain if functional assays are not calibrated.
To address this, our lab created a calibrated functional assay that calculates an odds of pathogenicity score for MSH2 and MSH6 gene variants that can be used as evidence for classifying variants of uncertain significance. This thesis focuses on the creation of human embryonic stem cell models of clinical variants with known pathogenicity using CRISPR technology, and microsatellite instability analysis as an assay to measure protein function. Methods of designing CRIPSR models and performing off-target analysis as a control step are described, and microsatellite instability was used to determine the corrective ability of mismatch repair machinery in detecting insertions and deletions in simple repeat sequences across the genome.
This calibrated assay provides critical information about variant function, and this statistical model can be used to assess variants that do not have enough supporting evidence to determine a pathogenicity classification. Additionally, Lynch syndrome individuals can better understand their risk of developing cancer and preventative methods that should be used to monitor potential tumorigenesis. For patients presenting with cancer, the pathogenicity of a mismatch repair variant may impact their treatment plan as well as preventative plans for them and their family members, so this functional evidence is critical in personalized medicine.
Recommended Citation
Amodeo, Olivia N., "The Development of a CRISPR-Based Calibrated Functional Assay for Classifying Pathogenicity of MSH2 and MSH6 Variants in Lynch Syndrome" (2026). Honors Scholar Theses. 1199.
https://digitalcommons.lib.uconn.edu/srhonors_theses/1199