Endocrine Abstracts

Background: Pheochromocytomas are rare tumours arising from adrenal medulla. Recent findings show that about 30–40% of pheochromocytomas are caused by germline mutations in one of the ten hereto known susceptibility genes: SDHA, SDHB, SDHC, SDHD, SDHAF2, RET, VHL, NF1, TMEM127 and MAX. This list of genes is constantly growing. These ten genes together consist of 128 exons and a genetic screening test is both extensive time-consuming and expensive. We introduce utilizing Next generation sequencing as a fast and cost effective method.

Methods: DNA was extracted from pheochromocytoma lesions and were subjected to whole genome sequencing utilizing Illumina Hi seq platform, performed at university core facility. Reads from pair-end fragments were mapped (GRCh37) and variation calling was performed using a commercially available software. Identified mutations were verified by automated Sanger sequencing.

Results: We have identified two point mutations, in RET and SDHC, in two different pheochromocytoma lesions, spending less than two hours hands on, in house, time. Mutations were verified by traditional Sanger sequencing of a single fragment for each gene.

Conclusions: Utilizing next generation sequencing is a fast and cost effective method for clinical genetic screening in pheochromocytomas. Using this workflow, the analysis can be performed without expert skills in bioinformatics.

Declaration of interest: The authors declare that there is no conflict of interest that could be perceived as prejudicing the impartiality of the research project.

Funding: This work was supported, however funding details unavailable.