Targeted sequencing panel for canine lymphoma

Targeted next-generation sequencing (NGS) enables the identification of genomic variants in cancer patients with high sensitivity at relatively low costs, and has thus opened the era to personalized human oncology. Veterinary medicine tends to adopt new technologies at a slower pace compared to human medicine due to lower funding, nonetheless it embraces technological advancements over time. Hence, it is reasonable to assume that targeted NGS will be incorporated into routine veterinary practice in the foreseeable future. Here, we present the TiHoCL targeted NGS panel, a molecular tool for characterizing and stratifying canine lymphoma (CL) patients. CL is a disease developing from uncontrolled and pathologic clonal expansion of lymphoid B- or T-cells. CL resembles human non-Hodgkin lymphoma (NHL) in many aspects. However, in contrast to NHL, CL is considered incurable: although over 80% of CL patients achieve complete remission after initial chemotherapy, most relapse within 12 months. Also, while various single nucleotide polymorphisms (SNPs) have been associated with high risk of developing NHL, poor prognosis and resistance to treatment in NHL patients, little is known about the genetics of CL. Accordingly, the molecular CL landscape is currently not efficiently utilized for advanced patient stratification. Still, because of the similarities between the two malignancies, it is conceivable to use knowledge about NHL to design a panel that could help guide treatment decisions in CL patients. Thus, the approximately 100 SNPs included in the TiHoCL targeted NGS panel were selected based on comparative genomics and network-based approaches, starting from a literature and database search for SNPs reported to be associated with both human and canine hematologic malignancies. The TiHoCL targeted NGS panel was technically and functionally evaluated by sequencing DNA samples isolated from blood of 29 lymphoma dogs using an Ion Torrent PGM System achieving good sequencing run metrics. Our design framework holds new possibilities for the design of similar molecular tools applied to other diseases for which limited knowledge is available. It serves as a proof of principle for target sequencing panels for veterinary medicine developed from human knowledge and will improve drug target discovery and patient care.