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Related Experiment Video

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Comparative Lesions Analysis Through a Targeted Sequencing Approach
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Comparative Lesions Analysis Through a Targeted Sequencing Approach.

Caterina Vicentini1, Andrea Mafficini1, Michele Simbolo2

  • 1ARC-Net Research Centre, University and Hospital Trust of Verona.

Journal of Visualized Experiments : Jove
|November 26, 2019
PubMed
Summary
This summary is machine-generated.

Understanding intra-tumoral heterogeneity (ITH) is crucial for cancer therapy. This study presents a method combining histology, sequencing, and immunophenotyping to analyze genetic relatedness between tumor lesions, identifying clonal and subclonal populations.

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Area of Science:

  • Oncology
  • Genomics
  • Cancer Research

Background:

  • Intra-tumoral heterogeneity (ITH) significantly impacts targeted therapy efficacy and treatment outcomes.
  • Understanding the genetic landscape of primary and metastatic lesions is critical for advanced-stage disease management.

Purpose of the Study:

  • To develop and validate a method for comparative analysis of distinct tumor lesions within a single patient.
  • To identify clonal and subclonal populations and assess genetic relatedness between primary and metastatic sites.

Main Methods:

  • Integration of histological analysis, high-coverage multi-lesion sequencing (targeting 409 cancer genes), and immunophenotypic analyses.
  • Neoplastic cell enrichment and careful pathological examination to minimize sample processing artifacts.
  • Comparative analysis of somatic alterations (SNVs, CNVs) and immunohistochemistry for functional prediction.

Main Results:

  • The developed method successfully distinguishes clonal from subclonal genetic alterations across different lesions.
  • The approach enables evaluation of heterogeneity in somatic alterations within distinct patient lesions.
  • Immunohistochemistry aids in predicting functional consequences of identified mutations.

Conclusions:

  • This integrated methodology provides a robust framework for assessing ITH and inter-lesion genetic relatedness.
  • The protocol is broadly applicable to various solid tumor types for improved therapeutic strategy design.
  • Accurate characterization of ITH is essential for anticipating therapy failure and optimizing anti-tumor strategies.