The myPath® Melanoma test is intended as an adjunctive diagnostic tool for melanocytic lesions that are difficult to classify by histopathology alone.
Using qRT-PCR technology, the test objectively distinguishes melanoma from benign nevi with greater than 90% accuracy in three independent clinical validations.
In the initial discovery phase of test development, 79 candidate genes were selected based upon published data demonstrating their differential expression in melanoma compared with benign nevi or their increased expression in aggressive tumors.9-17
This panel was refined to a set of the 40 genes that most effectively differentiated benign and malignant melanocytic lesions, and these genes were then further assessed in a training cohort of archival formalin-fixed paraffin-embedded melanocytic lesions (n = 464).
Housekeeping genes included: CLTC, MRFAPI, PPP2CA, PSMA1, RPL13A, RPL8, RPS29, SLC25A3, and TXNLI
Statistical modeling identified a subset of 14 genes grouped into three distinct gene components which provided the greatest sensitivity and specificity. These 14 signature genes are involved in cell differentiation and cellular immune signaling.
Expression of the signature genes is normalized to that of 9 housekeeper genes prior to the application of a weighted algorithm that combines the measurements of all signature genes and generates a single numerical score.
Validation Study 1:
- Retrospective cohort (n=437) representing broad range of clinical and histopathologic subtypes (entirely separate from training cohort)
- Reference standard: Independent concordant diagnosis by 2 expert dermatopathologists
- Sensitivity = 90%; Specificity = 91%15
Validation Study 2:
- Prospective cohort (n=1,172) of cases submitted for testing in the clinical setting
- Reference standard: Independent concordant diagnosis by 3 expert dermatopathologists
- Diagnostic concordance among all 3 in 736 cases
- Sensitivity = 92%; Specificity = 93%17
- Included ambiguous / diagnostically equivocal cases; expert panelists documented uncertainty in >20% of the 736 cases (e.g., “indeterminate case,” “borderline tumor,” “requires ancillary studies,” “differential diagnosis includes nevus and melanoma,” “re-excise to exclude melanoma,” etc.)
Validation Study 3:
- Retrospective cohort (n=182) with clinical outcomes
- 99 melanomas that developed documented distant metastasis after initial biopsy
- 83 nevi with median event-free follow-up > 6 years
- Reference standard: Patient outcomes (distant metastasis or ≥ 5 year event-free follow-up)
- Sensitivity = 94%; Specificity = 96%18
PRAME encodes a cancer-testis protein19 that is aberrantly expressed in melanoma. It appears to contribute to tumorigenesis by functioning as a dominant repressor of retinoic acid receptor signaling20 and / or down-regulation of TRAIL expression.21
The second component contains five genes from the S100A family: S100A7, S100A8, S100A9, S100A12, and PI3. The products of these genes are involved in multiple cellular processes. S100A9 is a calcium binding protein often found in combination with S100A8 as part of an immunogenic protein heterodimer.22 Increased S100A8 and S100A9 levels are detected in many malignant neoplasms,23-25 both within tumor cells and within infiltrating immune cells.
The third component contains 8 genes involved in tumor immune response signaling: CCL5, CD38, CXCL10, CXCL9, IRF1, LCP2, PTPRC, and SELL. Many of these genes produce chemokines or chemokine receptors that regulate leukocyte trafficking. Chemokines can suppress or promote the growth of a neoplasm by acting on cells of the tumor microenvironment, including leukocytes, endothelial cells, and fibroblasts, but they may also affect tumor cells themselves by regulating migration, invasion, proliferation, and resistance to chemotherapy.26
The fourth component is a group of nine housekeeping genes whose measurement allows normalization of the RNA expression for analysis.