Genomics Learning Center

Somatic Variant Analysis & Tumor Profiling

A deep-dive guide to somatic variant detection, AMP-guided classification, and the evolving landscape of precision oncology diagnostics.

Precision medicine aims to transform healthcare by utilizing an individual's genetic makeup and clinical presentation to aid in diagnosis, treatment, and prevention. In the cancer space, data derived from next-generation sequencing (NGS) is used to diagnose and prognose diseases, select targeted therapies, and evaluate a patient's suitability for clinical trials.

This growth creates a significant challenge: labor-intensive diagnostic processes that require deep expertise are meeting rapidly increasing demand. Clinical laboratories can expect to multiply their workload within a few years while facing a shortage of clinical genetics experts with NGS-specific training. Automation of variant interpretation and reporting is the only viable path forward.

Somatic vs. Germline Cancer Variants

Cancer can be triggered by both germline and somatic variants. Germline variants occur in germ cells (egg or sperm) and are hereditary, such as mutations in BRCA1 and BRCA2. Somatic variants occur in any other cell type after birth, driven by environmental effects or DNA replication errors.

Both types can be activating (conferring new or increased cell activity that promotes tumor development) or inactivating (causing loss of function that inhibits tumor suppression). For example, mutated BRCA1 switches off a tumor-suppressor gene, and the homozygous loss of TP53 is frequently observed in colon, breast, and lung cancers.

Clinically Relevant Cancer Variant Types
SNVs
Single-Nucleotide Variants
e.g., BRAF V600E in melanoma
Indels
Small Insertions or Deletions
e.g., frameshift mutations in BRCA2
CNVs
Copy Number Variants
e.g., ERBB2 amplification; RB1 loss in retinoblastoma
Fusions
Gene Fusions
e.g., BCR-ABL1 in chronic myeloid leukemia
Splice
Splice Site Variants
e.g., MET exon 14 skipping in NSCLC

The AMP/ASCO/CAP Guidelines

The key standards for somatic variant interpretation were published by Li et al. (2017) as a joint consensus of the Association for Molecular Pathology (AMP), American Society of Clinical Oncology (ASCO), and College of American Pathologists (CAP). Unlike germline interpretation (which focuses on pathogenicity), somatic variant assessment evaluates drug sensitivity, drug resistance, and potential toxicity.

Evidence Levels

A
FDA-Approved / Professional Guidelines
Establishes tumor response or resistance for a specific tumor type
B
Strong Evidence / Expert Consensus
Large studies with expert consensus on clinical interpretation
C
Different Tumor Type Evidence
Used for off-label recommendations or clinical trial enrollment
D
Pre-Clinical / Small-Scale
Pre-clinical studies or smaller-scale evidence

Tier Classification

I
Strong Clinical Significance
Level A/B evidence — e.g., BRAF V600E predicting vemurafenib response
II
Potential Clinical Significance
Level C/D evidence — off-label use or clinical trial inclusion
III
Unknown Clinical Significance
Rare variants with potentially damaging impact but no conclusive association
IV
Benign
Common variants ruled out by population allele frequency

Immunotherapy Biomarkers

Modern oncology testing goes beyond individual variants. Three key biomarkers have become standard in tumor profiling for identifying patients eligible for immunotherapy and targeted treatments.

Tumor Mutational Burden (TMB)

The number of somatic mutations per megabase. High TMB correlates with increased neoantigens and more robust T-cell activation. Pembrolizumab received FDA approval for advanced solid tumors with TMB >10 mut/Mb.

Microsatellite Instability (MSI)

Results from failure of mismatch repair proteins to fix DNA replication errors, leading to frameshift mutations. Associated with colon, gastric, endometrial, and ovarian cancers. MSI-high tumors respond to checkpoint immunotherapy.

Homologous Recombination Deficiency (HRD)

A deficiency in the HRR DNA repair pathway associated with breast, ovarian, prostate, and pancreatic cancers. HRD makes tumors more sensitive to platinum-based therapies and PARP inhibitors.

The Somatic Analysis Workflow

Somatic variant analysis requires distinguishing true oncogenic drivers from germline background and sequencing artifacts. A structured environment for high-sensitivity detection and evidence-based classification is essential.

1

Detection & Sensitive Filtering

Calculate allelic ratios and VAFs to identify low-frequency mutations. Filter against population databases (gnomAD, ExAC) to exclude rare germline variants and apply lab-specific blacklists to remove recurring artifacts.

2

Oncogenicity Scoring

Prioritize driver mutations using scoring algorithms that integrate COSMIC recurrence, in-silico functional predictions (SIFT/PolyPhen/CADD), and splice-site disruption analysis.

3

AMP Tier Classification

Transition from analysis to interpretation using AMP/ASCO/CAP guidelines. Automate the assignment of Tiers I–IV based on evidence levels A–D, connecting variants to professional guidelines and therapeutic insights.

4

Clinical Reporting

Generate structured reports with curated cancer interpretations, FDA-approved therapy matches, and active clinical trial recommendations based on the patient's genomic profile.

AMP Tier Classification
Sample ID: TUM-4421
BRAF p.V600E
VAF: 42.4% | Reads: 1,120x
Tier I — Strong
Evidence Category
Level A (Therapeutic)
Guideline Recommendation

FDA-approved therapy (Vemurafenib) indicated for patients with BRAF V600E mutation in melanoma.

Oncogenicity
0.99
Hotspot Status
Positive

Key Annotation Sources & Databases

Clinical cancer analysis relies on integrating multiple categories of databases. Interpretation is only as good as the underlying data.

Population Databases

gnomAD, dbSNP, ExAC, 1000 Genomes — used to exclude common germline variants from somatic analysis.

Cancer Databases

COSMIC, CIViC, Cancer Hotspots, TCGA — cataloging somatic mutations with tumor histology and clinical associations.

Therapy Databases

DrugBank, NCI Thesaurus, ClinicalTrials.gov — linking biomarkers to FDA-approved therapies and active clinical trials.

Golden Helix CancerKB™

Professionally curated knowledgebase with report-ready interpretations for thousands of somatic biomarkers.

Challenges in Cancer Genomics

While the field has advanced rapidly, several key challenges remain for clinical laboratories performing somatic variant analysis.

Detection Sensitivity

Somatic mutations may occur in only 1–5% of sequence reads due to tumor heterogeneity and biopsy composition. Sensitivity down to 10% of tumor cells is desirable.

Tumor-Only Testing

Many tests sequence only tumor tissue without matched normal, limiting the ability to distinguish somatic mutations from rare germline variants at novel sites.

Reporting Inconsistency

Surveys show substantial variation in what is reported across labs — only 35–37% report CNVs and fusions, and labs use different classification schemes.

Rapidly Evolving Knowledge

New cancer-gene associations, treatments, and clinical trials are published daily. Tier classifications are snapshots in time — Tier II variants may become Tier I as evidence grows.

Cancer Genomics Insights & Webcasts

Expert insights into AMP guidelines, tumor profiling, and the evolving landscape of cancer genomics.

Ready to Master Somatic Analysis?

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AMP Classification
Tumor-Normal & Tumor-Only
Somatic CNV Detection