Comparison of gene fusion detection methods in salivary gland tumors

• Published in: Human pathology 2022-05, Vol.123, p.1-10
• Main Authors: Sun, Lulu, Petrone, Jessica S., McNulty, Samantha N., Evenson, Michael J., Zhu, Xiaopei, Robinson, Joshua A., Chernock, Rebecca D., Duncavage, Eric J., Pfeifer, John D.
• Format: Article
• Language: English
• Subjects: Adenoma, Pleomorphic - pathology; Cancer; Carcinoma - pathology; Carcinoma, Acinar Cell - pathology; Exocrine glands; Fluorescence in situ hybridization; Gene Fusion; Gene fusions; Genes; Genomics; Humans; In Situ Hybridization, Fluorescence - methods; Laboratories; Molecular pathology; Oral cancer; RNA sequencing; Salivary Gland Neoplasms - diagnosis; Salivary Gland Neoplasms - genetics; Salivary Gland Neoplasms - pathology; Salivary gland tumors; Tumors
• ISSN: 0046-8177; 1532-8392
• DOI: 10.1016/j.humpath.2022.02.002

Salivary gland neoplasms may pose diagnostic difficulties due to overlapping morphologic features. Recently, specific gene fusions have been discovered that correspond to particular tumor types, and can aid in accurate diagnosis. Gene rearrangements are commonly assessed by fluorescence in situ hybridization (FISH), although use of next-generation sequencing is increasing. However, there is no “gold standard” for fusion detection. We determined the concordance between FISH and a targeted RNA sequencing panel in gene fusion detection across twenty-two salivary gland tumors, including five mucoepidermoid carcinomas, four acinic cell carcinomas, four pleomorphic adenomas, two adenoid cystic carcinomas, two NUT carcinomas, and one each of basal cell adenoma, salivary duct carcinoma ex-pleomorphic adenoma, salivary duct carcinoma, clear cell carcinoma, and secretory carcinoma. Directed FISH testing based on the diagnosis was performed on cases that did not already have FISH conducted during clinical workup. Targeted RNA sequencing of 507 genes and their partners (using the Illumina TruSight Fusion Panel) was completed. Six of twenty-two (27.3%) cases had discordant results. In three cases, FISH results were negative while RNA sequencing results found fusion transcripts, which were all confirmed with RT-PCR and Sanger sequencing. In three cases, RNA sequencing results were negative while FISH results were positive for a gene rearrangement. Thus, if fusion analysis results are conflicting with the morphologic impression, a second mode of fusion detection may be warranted. Although both methods have advantages and drawbacks, RNA sequencing provides additional information about novel fusion partners and fusions that may not have been originally considered. •Detection of characteristic gene fusions is useful in the diagnosis of salivary gland tumors.•Targeted RNA sequencing and fluorescence in situ hybridization can be used to identify fusions.•More than a quarter of cases examined had discordant results between methods.•Both methods can provide false negative results.