Targeted deep sequencing of CD34+ cells from peripheral blood can reproduce bone marrow molecular profile in myelodysplastic syndromes

Martin, Roman; Acha, Pamela; Ganster, Christina; Palomo, Laura; Dierks, Sascha; Fuster‐Tormo, Francisco; Mallo, Mar; Ademà, Vera; Gomez-Marzo, Paula; Haro, Nuri De; Solanes, Neus; Zamora, Lurdes; Xicoy, Blanca; Shirneshan, Katayoon; Flach, Johanna; Braulke, Friederike; Schanz, Julie; Kominowski, Arkadiusz; Stromburg, Martin; Brockmann, Alina; Trümper, Lorenz; Solé, Françesc; Haase, Detlef

doi:10.1002/ajh.25089

Cited by 13 publications

(8 citation statements)

References 5 publications

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“… 4,23 The potential use of PB for the reproduction of BM molecular profiles was previously demonstrated by the application of NGS- or fluorescence in situ hybridization–based analysis on immunomagnetic enriched CD34 + cells from PB. 24,25 In line with previous observations, 11 our data indicate that the sensitivity for MRD detection (using both DC and NGS) in CD34 + -sorted PB samples is at least equal to or even higher than that in CD34 + cells from BM aspirates. This finding may indicate that a higher proportion of malignant CD34 + cells (vs normal CD34 + cells) are released into PB circulation from the BM environment, and/or a delayed clearance of leukemic blasts in peripheral circulation occurs after treatment.…”

Section: Discussionsupporting

confidence: 92%

Deep sequencing in CD34+ cells from peripheral blood enables sensitive detection of measurable residual disease in AML

et al. 2022

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Monitoring of measurable residual disease (MRD) in patients with acute myleoid leukemia (AML) is predictive for disease recurrence and may identify patients who benefit from treatment intensification. Current MRD techniques rely on multicolor flow cytometry or molecular methods, but are limited in applicability or sensitivity. We evaluated the feasibility of a novel approach for MRD detection in peripheral blood (PB), which combines immunomagnetic preenrichment and fluorescence-activated cell sorting (FACS) for isolation of CD34+ cells with error-reduced targeted next-generation sequencing (NGS). For clinical validation, we retrospectively analyzed 429 PB and 55 bone marrow (BM) samples of 40 AML and high-risk MDS patients, with/without molecular relapse based on CD34+ donor chimerism (DC), in complete remission after alloHSCT. Enrichment of CD34+ cells for NGS increased the detection of mutant alleles in PB ~1000-fold (median VAF 1.27% vs 0.0046% in unsorted PB; P<0.0001). Although a strong correlation was observed for the parallel analysis of CD34+ PB cells with NGS and DC (r=0.8601), the combination of FACS and NGS improved sensitivity for MRD detection in dilution experiments ~10-fold to levels of 10-6. In both assays, MRD detection was superior using PB versus BM for CD34+ enrichment. Importantly, NGS on CD34+ PB cells enabled prediction of molecular relapse with high sensitivity (100%) and specificity (91%), and significantly earlier (median 48 days, range 0-281; P=0.0011) than by CD34+ DC or NGS of unsorted PB, providing additional time for therapeutic intervention. Moreover, panel sequencing in CD34+ cells allowed the early assessment of clonal trajectories in hematological complete remission.

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Section: Discussionsupporting

confidence: 92%

Deep sequencing in CD34+ cells from peripheral blood enables sensitive detection of measurable residual disease in AML

et al. 2022

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“…Our data demonstrate that the analysis of cfDNA represents a novel strategy that would be useful for routine testing, as cfDNA is obtained fast and easily from blood plasma, when compared with BM aspirates or purified CD34 + cells. 19 …”

Section: Discussionmentioning

confidence: 99%

Molecular and cytogenetic characterization of myelodysplastic syndromes in cell-free DNA

et al. 2022

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Molecular and cytogenetic studies are essential in patients with myelodysplastic syndromes (MDS) for diagnosis and prognosis. Cell-free DNA (cfDNA) analysis has been reported as a reliable non-invasive approach for detecting molecular abnormalities in MDS, however, there is limited information about cytogenetic alterations and monitoring in cfDNA. We have assessed the molecular and cytogenetic profile of a cohort of 70 patients with MDS by next-generation sequencing (NGS) using cfDNA and compared the results to paired bone marrow (BM) DNA. Sequencing of BM DNA and cfDNA showed a comparable mutational profile (92.1% concordance) and variant allele frequencies (VAF) strongly correlated between both sample types. Of note, SF3B1 mutations were detected with significantly higher VAF in cfDNA than in BM DNA. NGS and microarrays were highly concordant to detect chromosomal alterations although with lower sensitivity than karyotype/FISH. Nevertheless, all cytogenetic aberrations detected by NGS in BM DNA were also detected in cfDNA. Additionally, molecular and cytogenetic alterations were monitored and we observed an excellent correlation between the VAF of mutations in BM DNA and cfDNA across multiple matched time points. A decrease in the cfDNA VAF was detected in patients responding to therapy, but not in non-responding patients. Of note, cfDNA analysis also showed cytogenetic evolution in 2 cases not responding to treatment. In conclusion, although further studies with larger cohorts are required, our results support the analysis of cfDNA as a promising strategy for performing molecular characterization, detection of chromosomal aberrations and monitoring of MDS patients.

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“…In addition, Mohamedali et al used SNP‐a and targeted gene sequencing of 24 genes methods to investigate the BM and PB concordance and their results showed 95% similarity. Martin et al compared the genetic results of bone marrow and immunomagnetically enriched CD34+ peripheral blood cells using targeted deep sequencing (TDS) method in MDS patients. According to their data, enrichment of CD34+ cells in peripheral blood can improve the detection of genetic abnormality up to 97% of blood samples.…”

Section: Discussionmentioning

confidence: 99%

Evaluation of the utility of peripheral blood vs bone marrow in karyotype and fluorescence in situ hybridization for myelodysplastic syndrome diagnosis

Fakhr

Mehrzad

Izaditabar³

et al. 2018

Clinical Laboratory Analysis

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Targeted deep sequencing of CD34+ cells from peripheral blood can reproduce bone marrow molecular profile in myelodysplastic syndromes

Cited by 13 publications

References 5 publications

Deep sequencing in CD34+ cells from peripheral blood enables sensitive detection of measurable residual disease in AML

Deep sequencing in CD34+ cells from peripheral blood enables sensitive detection of measurable residual disease in AML

Molecular and cytogenetic characterization of myelodysplastic syndromes in cell-free DNA

Evaluation of the utility of peripheral blood vs bone marrow in karyotype and fluorescence in situ hybridization for myelodysplastic syndrome diagnosis

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