Effectiveness and Impact of Transcript Analysis in Clinical Genetics Daily Practice

Area of Science:

• Clinical Genetics and Genomics
• Molecular Diagnostics
• RNA Splicing Analysis

Background:

• Broad genetic testing frequently identifies variants of uncertain significance (VUS).
• Interpreting VUS that potentially affect RNA splicing is a significant diagnostic challenge.
• Accurate VUS classification is crucial for patient diagnosis and management.

Purpose of the Study:

• To develop and validate a method for classifying splicing-associated VUS.
• To integrate peripheral blood mRNA transcript analysis into routine clinical diagnostics.
• To assess the clinical impact of reclassifying VUS through transcript analysis.

Main Methods:

• Collected peripheral blood samples from patients with VUS in specific genes.
• Performed mRNA transcript analysis to detect splicing alterations.
• Integrated transcript data with existing genetic and phenotypic information for VUS classification.

Main Results:

• Identified splicing alterations caused by VUS in DICER1, MSH2, MLH1, DYNC1H1, RPS6KA3, and SCN9A.
• Reclassified identified VUS as Pathogenic or Likely Pathogenic based on splicing impact.
• Demonstrated significant clinical utility for diagnosing hereditary tumor, neurological, and congenital disorders.

Conclusions:

• Transcript analysis is a valuable tool for evaluating splicing-associated VUS.
• Incorporating RNA analysis into diagnostic workflows enables timely and accurate VUS classification.
• This approach improves clinical decision-making and patient care for genetic disorders.