Table 4 Performance of MRD monitoring by IGH/K rearrangement in different B cell malignancies

Genuardi et al. [96]

MCL(20)

BM(10) or

PB(10)

Phase III MCL0208

95% (19/20)

Sanger sequencing: 75% (15/20)

87% (13/15)

Not available

Not available

Not available

NGS-based IGH screening might have the ability to track major clones in MRD monitoring

Ladetto et al. [91]

ALL(15), MCL(30), MM(10)

BM(218) or

PB(160)

Prospective clinical trials

ALL: 100%(15/15)

MCL: 86%(26/30)

MM: 80%(8/10)

ASO-PCR:

ALL:100%(15/15)

MCL: 73%(22/30)

MM: 80%(8/10)

95.5%(41/43)

Not available

ASO-PCR, not available

Fully concordant: 79.6% (211/265)

Discordant: 20.4%(54/265), with 1.5% (4/265) major qualitative discordance, 5.3%(36/265) borderline qualitative discordance and 5.3%(14/265) quantitative discordance

NGS used in the identification of IGH clonotypes provides results that are at least comparable to ASO-PCR

Pulsipher et al. [122]

ALL(56)

BM

(41 for pre-HCT analysis, 125 for post-HCT MRD)

Trial ASCT0431

100% (41/41)

Not available

Not available

Relapse probability is 0% (0/22) and 53% (9/19) for pre-HCT NGS-MRD- and pre-HCT NGS-MRD + patients, respectively

Relapse probability is 25% and 67% for post-HCT NGS-MRD- and post-HCT NGS-MRD + , respectively

FC: Relapse probability is 16% and 46% for pre-HCT MFC-MRD- and pre-HCT MFC-MRD + patients

11 patients with post-HCT NGS-MRD + and post-HCT MFC-MRD- relapsed; none of patients with post-HCT NGS-MRD- and post-HCT MFC-MRD + relapsed

IGH V(D)J NGS-MRD predicted relapse and survival more accurately than FC-MRD

Ho et al. [19]

MM(251)

BM(438)

Treated at MSKCC

93.6% (235/251)

EC and Sanger sequencing: 93.6%

100%

78.6% (147/187) of the MRD samples with an IG NGS-MRD + status

81.8% (153/187) of the MRD samples with an hsFC-MRD + status

concordance of 92.9% (170/183) in MRD status detected by NGS and hsFC

NGS and hsFC performed similarly, showing a high concordance rate

Medina et al. [21]

MM(106)

BM()

Spanish GEM2012 clinical trial

Not available

Not available

Not available

50% (53/106) of patients with an IG NGS-MRD- status

54.7% (58/106) of patients with an NGS-MRD- status

Good correlation between the two methods (r = 0.951, R² = 0.905) with 15 discordant cases (5NGF + /NGS-; 10 NGF-/NGS +)

NGS has the excellent applicability and comparable results to NGF

Avet-Loiseau et al. [22]

MM(1085)

BM

Phase 3 CASSIOPEIA study

Not available

Not available

Not available

344 patients achieved an IG NGS-MRD- status

582 patients achieved a MFC-MRD- status

Good overall agreement was achieved in 83.5% of 733 patients evaluated by both NGS and MFC

NGS and NGF perform similarly in evaluating MRD regardless of response and CR status

Li et al. [121]

ALL(258)

BM or PB (258)

Ma-Spore ALL 2003 and ALL 2010 studies

497 disease clones in 90.3% (233/258) patients

Sanger Sequencing: 348 disease clones in patients

90.8% of clones detected by Sanger sequencing were identified by IG NGS

78% (54/69) of samples with quantifiable MRD detected by IG NGS

58% (40/69) of samples with quantifiable MRD detected by RQ-PCR

40/69 of samples with quantifiable MRD detected by both IG NGS and RQ-PCR, 15/69 of samples with negative MRD detected by both methods

Sub-clonal disease can be uncovered by IGH NGS compared with Sanger sequencing; IGH NGS shows improved sensitivity compared with RQ-PCR

Kriegsmann et al. [16]

MM(125)

BM(125 pairs)

Multi-centre prospective phase III HD6 trial

Not available

Not available

Not available

74.4% (93/125) of patients had an IG NGS- MRD + status

48% (60/125) of patients had a FC-MRD + status

68% (85/125) cases exhibited concordant MRD status detected by IG NGS and MFC

There exists good concordance between NGS and FC at a threshold of 10^–5

Langerhorst et al. [116]

MM(41)

BM(NGS, 81

Or

PB(MS, 82)

IFM-2009 clinical trial

Not available

Not available

Not available

18.5%(15/81) of samples were IG NGS-MRD-

21% (17/82) of samples were MS-MRD-

79% (64/81) of paired samples showed concordant MRD status detected by IG NGS and MS

MS is at least as sensitive to detect MRD compared with NGS and is alternative to NGS-MRD

Takamatsu et al. [12]

MM(125)

BM(125)

High-dose melphalan plus ASCT

An overall clone identification rate of 90% (113/125) by IG NGS method

An overall clone identification rate of 66% (75/113) by ASO-PCR method

Not available

Not available

ASO-PCR, not available

35 samples are NGS-MRD + /ASO-PCR-MRD- status;

Patients with IG NGS-MRD + /ASO PCR-MRD- status (11) showed worse PFS than patients with IG NGS-MRD- status (7)

Low level MRD detected by NGS but not ASO-PCR has significant prognostic value

Yao et al. [23]

MM(4)

BM(11)

VTD/PAD induction + ASCT + thalidomide maintenance

Disease clones were detected by IG NGS in 100% (4/4) of diagnostic samples

Disease clones were detected by ASO-PCR and Sanger sequencing in 100% (4/4) of diagnostic samples

Disease clones detected by the two methods were 100% same

5 samples achieved MRD + status and

2 samples achieved MRD- status by IG NGS

5 samples achieved MRD + status and

2 samples achieved MRD- status by ASO-PCR

100% of the 7 follow-up samples achieved a concordant MRD status detected by IG NGS and ASO-PCR method

NGS yields MRD measurements concordant and comparable to ASO-PCR; NGS shows improved sensitivity

Medina et al. [24]

MM(101)

BM

GEM2012 MENOS65 clinical trail

Clonality was confirmed in 100% (101/101) of cases with IG NGS

Clonality was confirmed in 99% (100/101) of cases with Sanger sequencing

97.9% (93/95) of the disease clones detected by IG NGS and Sanger sequencing were concordant

Not available

NGF, not available

High correlation (R² > 0.8) was maintained between NGF and NGS performed in each center,

Only 14% (13/93) of cases were discordant: 4 NGS-MRD- and NGF-MRD + cases, 9 NGS-MRD + and NGF-MRD- cases

NGS is a suitable strategy for clonality and MRD detection with results comparable to gold standards (NGF and Sanger sequencing)