Defining accuracy and sensitivity of T-cell receptor repertoire profiling for monitoring recurrent/persistent disease in patients with mature T-cell neoplasms (TECH1P.857)

Accurate and sensitive identification of recurrent or persistent disease in T-cell neoplasms is important for proper patient care. We developed a method that amplifies rearranged TCR CDR3 sequences and uses high-throughput sequencing (HTS) to sequence tens of thousands of chains simultaneously. Because the technology utilizes gDNA the frequency of sequenced CDR3 chains is representative of the relative frequency of each CDR3 sequence in the sample population of T cells. Thus these assays can describe both the breadth of the T-cell receptor repertoire and quantify individual clones; enabling tracking the presence and frequency of neoplastic clones. To demonstrate the potential of this technology, we first test the assay’s sensitivity and accuracy (1 lymphocyte in 1M total cells). Then, to show utility, we apply the technology to monitor persistent/recurrent disease in a set of mature T-cell lymphoma samples. To identify the neoplasm’s CDR3 chain, we sequence the TRB and TRG repertoire of 35 index samples. Then, to diagnose persistent disease, we sequence the TCR repertoire of follow-up samples and search for the neoplastic sequence. We then define persistent disease based on the sensitivity and accuracy of the assay. Concurrently, as part of standard clinical practice, a number of other methods were used to identify persistent disease. We compare these two methods to demonstrate that HTS technology is a viable alternative to traditional methods to detect and monitor lymphoma.