Assessment of Clonotypic Rearrangements and Minimal Residual Disease in Lymphoid Malignancies

Abstract

Context.— Measurable (minimal) residual disease (MRD) is an independent prognostic factor for survival outcomes in patients with lymphoid and plasma cell malignancies and has been incorporated into consensus criteria regarding treatment response, strategy, and clinical trial endpoints. clonoSEQ (a next-generation sequencing [NGS]-MRD assay) uses multiplex polymerase chain reaction and NGS to identify clonotypic rearrangements at the immunoglobulin (Ig) H, IgK, IgL, T-cell receptor (TCR)-β, and TCR-γ loci, and translocated B-cell lymphoma 1/IgH and 2/IgH sequences for MRD assessment. Additionally, it can be used to confirm diagnoses of cutaneous T-cell lymphoma (CTCL). Objective.— To review the technical aspects of our experience using the clonoSEQ Assay in routine clinical practice. Design.— In this single-center experience, 390 patients with lymphoid and plasma cell malignancies were assessed with the NGS-MRD Assay at a central laboratory. Results.— Median time from arrival of the shipment to initiation of the assay (defined as captured in Adaptive's secure tracking system) was 2.1 hours. Overall, 317 patients had 1 or more samples submitted for sequence identification. Of these, 290 (91.5%) had trackable sequences identified. The median calibration rate of samples by malignancy (where n ≥ 10 samples, excluding CTCL samples) was 88.1%, across a variety of fresh and archived sample sources (177 of 201 samples). TCR-β and/or TCR-γ clonotypes were identified in 40 of 95 samples (42.1%) from 66 patients with suspected CTCL. Conclusions.— This NGS-MRD Assay is a valuable and sensitive tool for monitoring MRD in patients with plasma cell and lymphoid malignancies and assisting in the diagnosis of CTCL.

Molecular techniques drive cutting edge advancements in management of cutaneous T cell lymphoma

“…In the same study, malignant T cells could be isolated from the blood of patients with new skin lesions but no clinical involvement of the peripheral blood, highlighting that early blood involvement may be detected by HTS. Numerous other studies have demonstrated that HTS has superior sensitivity and/or specificity as compared to PCR for detection of malignant cells in the skin and or blood ( 35 – 38 ).…”

Integrated antibody and cellular immunity monitoring are required for assessment of the long term protection that will be essential for effective next generation vaccine development

“…ScRNA analysis can also be used to identify multi-functional responding cells (35, 36). TCR sequencing focuses on the antigen receptors expressed by T cells and has been used for diagnosis and personalized monitoring of remission and relapse in multiple hematologic cancers (32,37). At the population level, TCR sequencing has been used to determine the clonality and repertoire dynamics of public T cell responses (38, 39).…”

Novel Approaches in Molecular Characterization of Classical Hodgkin Lymphoma

“…Potentially clinically relevant applications of cfDNA involve the detection of minimal residual disease (MRD) to monitor therapy response, as has been applied in different lymphoma types [80,81], including cHL [50,82], and for prognostic stratification [14,52,[82][83][84][85]. This requires sensitive NGS-based assays, such as ClonoSEQ (formerly known as Lym-phoSIGHT) [81,86], CAPP-Seq [87], PhasED-Seq [88], or the EuroClonality-NGS DNA capture (EuroClonality-NDC) assay [89].…”