Targeted genotyping of circulating tumor DNA for classical Hodgkin lymphoma monitoring: a prospective study

Abstract

The relevance of circulating tumor DNA (ctDNA) analysis as a liquid biopsy and minimal residual disease tool in the management of classical Hodgkin Lymphoma (cHL) patients was demonstrated in retrospective settings and remains to be confirmed in a prospective setting. We developed a targeted Next-Generation sequencing (NGS) panel for fast analysis (AmpliSeq® technology) of nine commonly mutated genes in biopies and ctDNA of cHL patients. We then conducted a prospective trial to assess ctDNA follow up at diagnosis and after 2 cycles of chemotherapy (C2). Sixty cHL patients treated by first line conventional chemotherapy (BEACOPPescalated [21.3%], ABVD/ABVD-like [73.5%] and other regimens [5.2%, for elderly patients] were assessed in this non-interventional study. Median age of the patients was 33.5 years (range 20-86). Variants were identified in 42 (70%) patients. Mutations of NFKBIE, TNFAIP3, STAT6,

Correlations between baseline 18F-FDG PET tumour parameters and circulating DNA in diffuse large B cell lymphoma and Hodgkin lymphoma

“…Concerning the burden parameters, TMTV was found to be significantly correlated with [cfDNA] and [ctDNA] levels in both DLBLC and cHL, as already found in previous studies [15,16]. We have also found that TMTS was frequently among the more highly correlated parameters with cfDNA and ctDNA levels (among the three first parameters in 2 cases of 4).…”

“…Thirty patients with diffuse large B cell lymphoma (DLBCL) and 50 patients with classical Hodgkin lymphoma (cHL) were enrolled prospectively. Description of the population with clinical characteristics and outcomes is available in two previous studies [15,16]. All patients had performed a PET/CT on one of 3 different machines (description summarized in Additional file 1: Table 1).…”

“…Circulating cell-free DNA ([cfDNA]) was extracted from 3 mL of plasma aliquots with Amp Circulating Nucleic Acid ® QI Kit (Qiagen, Hilden, Germany) according to the manufacturer's instructions and quantified using QuBit High Sensitivity dsDNA (ThermoFisher Scientific, Illkirch, France). After tumour sequencing adapted to the type of cancer [15,16], the DNA was eluted in 60 to 80 μL of AVE buffer and then stored at − 80 °C. Quantification of the double-stranded DNA was performed by fluorometry on Qubit 2.0 (ThermoFisher Scientific Carlsbad, CA, USA), with Qubit ® dsDNA kit HS Assay (ThermoFisher Scientific, Carlsbad, CA, USA).…”

“…In these previous studies, [ctDNA] was correlated with factors of poor prognosis, notably international pronostic index (IPI) for DLBCL and stage 3-4, anaemia and symptoms B for cHL. Moreover high TMTV and high [ctDNA] level were found to be significantly correlated at the time of diagnosis for both DLBCL and cHL [15,16]. However, these correlations were moderate, suggesting that other factors may influence these two approaches of tumour burden quantification.…”

“…In cancerous patients, plasma ctDNA is released by apoptotic tumour cells, by necrotic tumour cells or is actively secreted by tumour cells [13] and ctDNA level can have a prognostic value at baseline, notably for DLBCL [14]. To quantify the ctDNA level ([ctDNA]), we have recently designed and validated two sets of genes for DLBCL (panel of 34 genes frequently mutated in DLBCL patients, so-called lymphopanel) [15] and for cHL (set of 9 genes) [16]. In these previous studies, [ctDNA] was correlated with factors of poor prognosis, notably international pronostic index (IPI) for DLBCL and stage 3-4, anaemia and symptoms B for cHL.…”