2023 ACMG Annual Clinical Genetics Meeting Digital Edition: Poster Gallery-Establishment of a novel functional assay to test ATM Variants of Uncertain Significance

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The document "Establishment of a novel functional assay to test ATM Variants of Uncertain Significance" discusses the development of a functional assay to evaluate the impact of uncertain ATM variants on cancer risk. The goal of this study is to establish accurate and reliable multiplexed assays to evaluate these variants. The researchers created a cell-based system to assess ATM variants and evaluated different functional assays to identify the best readout for ATM function.<br /><br />The researchers used a PROTAC-based approach to rapidly and controlled degrade ATM in cells. They generated HEK293 cell lines expressing ATM fused with FKBP12F36V using CRISPR-CAS9 and introduced ATM variants to test the system. The assay output was not influenced by cellular adaptation and provided a quantitative multifaceted readout.<br /><br />The document also provides background information on the increase in variants of unknown significance (VUS) due to increased genetic testing to identify cancer-associated mutations. The ATM gene, a moderate cancer risk gene, encodes for a kinase critical for DNA double-strand break response, cell-cycle checkpoint, and apoptosis. Working with ATM is challenging due to its complex structure and large gene size with highly conserved domains.<br /><br />The document includes images and results from the study, including the characterization of cell lines expressing ATM-FKBP12F36V-HA protein, ATM degradation and recovery after washout, and phosphorylation events after irradiating cells treated with dTAGv1.<br /><br />In summary, this study aims to establish a functional assay to evaluate uncertain ATM variants and provides insights into the development of a cell-based system using a PROTAC-based approach. The results demonstrate the potential of this assay to determine the functional impact of ATM variants and improve understanding of their cancer risk association and clinical management.

Asset Subtitle

Presenting Author - Ishor IT. Thapa, MS; Co-Author - Kathleen Clark, PhD; Co-Author - Sean V. Tavtigian, PhD;

Meta Tag

Genomic Methodologies

Genotype-Phenotype Correlations

Phenotype

Co-Author Kathleen Clark, PhD

Co-Author Sean V. Tavtigian, PhD

Presenting Author Ishor IT. Thapa, MS

Keywords

ATM variants

functional assay

cancer risk

multiplexed assays

cell-based system

PROTAC-based approach

HEK293 cell lines

CRISPR-CAS9

variants of unknown significance

ATM gene