PMCID string | Title string | Sentences string |
|---|---|---|
PMC12317575 | Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study | Within 24 h, cellular fractions were stored in Trizol (Invitrogen, Carlsbad, CA) at −80 °C (the Netherlands) or transferred to PAXgene blood RNA tubes (QIAGEN, Venlo, the Netherlands) and stored at −20 °C (Austria). |
PMC12317575 | Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study | RNA isolation and RT-qPCR were performed in Amsterdam, the Netherlands, while AIPF analysis took place in Vienna, Austria. |
PMC12317575 | Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study | Bone marrow histology and cytomorphology were centrally reviewed. |
PMC12317575 | Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study | Cytomorphological (CM) examination of Wright-Giemsa-stained smears was performed independently at St. Anna Children’s Hospital (Vienna, 10% cutoff) and the Princes Maxima Centre (Utrecht, the Netherlands, 1% cutoff), following respective study protocols. |
PMC12317575 | Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study | CM and histology were reported as either “positive”, “negative”, “inconclusive”, “dubious”, or “suspicious”. |
PMC12317575 | Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study | In this study, inconclusive and dubious samples were classified as negative, while suspicious samples were considered positive. |
PMC12317575 | Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study | mRNA extraction, complementary DNA (cDNA) synthesis and RT-qPCR followed established protocols . |
PMC12317575 | Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study | mRNA was isolated from PAXgene blood RNA tubes (QIAGEN) using the PAXgene Blood RNA Kit (QIAGEN), or from cellular fractions stored in Trizol (Invitrogen), per manufacturer instructions. |
PMC12317575 | Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study | RNA concentration and quality were measured using an ND-1000 spectrophotometer (Nanodrop). |
PMC12317575 | Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study | cDNA synthesis was done using the High-Capacity RNA-to-cDNA™ Kit (Applied Biosystems, Foster City, CA, USA) . |
PMC12317575 | Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study | RT-qPCR was performed on the Viia7 (Applied Biosystems, Carlsbad, CA, USA) and analyzed using QuantStudio software v1.6 (Applied Biosystems). |
PMC12317575 | Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study | The housekeeping gene glucuronidase beta (GUSB) and the neuroblastoma-specific mRNA marker PHOX2B were used as single markers . |
PMC12317575 | Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study | ADRN markers (TH, CHRNA3 and GAP43), and MES markers (POSTN, PRRX1 and flavin-containing monooxygenase 3 (FMO3)), were performed in multiplex panels . |
PMC12317575 | Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study | mRNA expression was normalized to GUSB (ΔCt = Ctmarker – CtGUSB). |
PMC12317575 | Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study | Samples with both GUSB Ct > 25 and PHOX2B negativity were excluded. |
PMC12317575 | Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study | A sample was scored positive if at least one of the markers scored positive, according to previously published thresholds . |
PMC12317575 | Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study | Level of mRNA detection (‘infiltration’) was calculated relative to neuroblastoma cell line IMR32 (2^ΔΔCT (ΔCTsample – ΔCTIMR32) * 100%). |
PMC12317575 | Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study | Median relative expression of positive markers was used to calculate infiltration level. |
PMC12317575 | Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study | Due to the calculations made relative to the expression in IMR32, calculated infiltrations can exceed 100%. |
PMC12317575 | Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study | For bilateral samples, the highest infiltration level was used. |
PMC12317575 | Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study | MES markers POSTN and PRRX1 were scored based on previously published thresholds relative to both GUSB and FMO3 expression . |
PMC12317575 | Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study | High expression of POSTN or PRRX1 and low expression of FMO3 identify MES neuroblastoma cells in bone marrow samples. |
PMC12317575 | Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study | Bone marrow aspirates were collected in EDTA tubes and subjected to density gradient centrifugation to isolate the mononuclear fraction, which also contains disseminated neuroblastoma cells. |
PMC12317575 | Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study | Cytospins (0.8–1.5 × 10 mononuclear cells (MNCs) per slide) were prepared, fixated and immunocytologically stained with labeled antibodies for GD2 and CD56 (Anti-GD2-FITC, Anti-CD56-Biotin, Mouse-Anti-Biotin-Cy3) (Supplemental Table 1A). |
PMC12317575 | Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study | Automated imaging (Axioplan 2, Zeiss, TRITC, FITC and DAPI filter) and analysis (Metafer 4, RCDetect classifier, Metasystems) identified DTCs . |
PMC12317575 | Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study | The total bone marrow MNC count per slide was extrapolated from representative measurements and GD2/CD56-positive cells were counted as DTCs. |
PMC12317575 | Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study | If fewer than four DTCs or ambiguous cells were detected, iFISH was performed using FISH probes (Metasystems) for primary tumor-specific aberrations (e.g., MYCN amplification, 1p deletion, 17q gain). |
PMC12317575 | Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study | Each analysis included positive and negative controls. |
PMC12317575 | Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study | If a sample was negative for both GD2 and CD56, no GD2- and/or CD56-positive tumor cells were detected. |
PMC12317575 | Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study | Infiltration was reported as the fraction of DTCs within MNC count . |
PMC12317575 | Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study | Detailed protocols are provided in Supplemental Data. |
PMC12317575 | Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study | Archived bone marrow cytopins from patient 166 with metastatic neuroblastoma were analyzed using imaging mass cytometry for twelve tumor-specific markers (antibodies against GD2, CD56/NCAM, CHGA, S100B, CXCR4, ELAVL4, GATA3, PRPH, SOX10, Vimentin, CD44, CD24) (Supplemental Table 1B), markers for nucleus detection and Ki-67 as a proliferation marker, as described in Lazic et al. (personal communication). |
PMC12317575 | Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study | Patient 166 was chosen due to the availability of multiple sequential timepoints in our study. |
PMC12317575 | Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study | Samples were thawed for 15 min at RT, fixed in 4% PFA (Carl Roth) at 4 °C for 30 min, washed twice in TBS and then blocked with 2% BSA (Carl Roth) and 0.1% Tween-20 (Merck) in TBS at RT for 1 h. Meanwhile, antibody cocktails were retrieved from −80 °C and thawed for 15 min at 4 °C before staining overnight at 4 °C. |
PMC12317575 | Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study | The following day, slides were washed twice (5 min per wash) in TBS, stained with the DNA intercalator Iridium (Fluidigm) and then dried with compressed air before IMC measurement with the Hyperion imaging system (Fluidigm) and CyTOF Software v7. |
PMC12317575 | Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study | Acquired images were background-corrected using spillover compensation with R CATALYST v.1.20.1 package , hot pixel removal through IMC-Denoise and semi-automated background correction . |
PMC12317575 | Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study | Nuclei were segmented using a Cellpose model fine-tuned in a human-in-the-loop approach with Cellpose 2.0 . |
PMC12317575 | Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study | Single-cell mean marker expressions were subsequently exported based on inferred segmentation masks. |
PMC12317575 | Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study | Statistical analysis was performed using SPSS v29 (RRID:SCR_002865; IBM Corporation, Armonk, NY, USA). |
PMC12317575 | Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study | Continuous variables were reported as median (10-90th percentiles), mean ± standard deviation, or mean (quartiles). |
PMC12317575 | Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study | Spearman correlations (95% confidence interval (CI)) assessed AIPF and RT-qPCR infiltration levels in paired samples. |
PMC12317575 | Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study | Independent samples t-test or Mann–Whitney U tests (for non-normal distributions) compared continuous variables. |
PMC12317575 | Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study | Graphs were generated using GraphPad Prism v9.3.1 (RRID:SCR_002798; GraphPad Software, Boston, Massachusetts, USA). |
PMC12317575 | Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study | One hundred eight (108) children with high-risk neuroblastoma, with a median age of 36 months at diagnosis (range 3–274 months) were included in this study (Table 1). |
PMC12317575 | Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study | Five hundred nine (509) bone marrow samples were collected (Fig. 1A & B), shipped and analyzed in specialized laboratories for AIPF and RT-qPCR. |
PMC12317575 | Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study | The current standard-of-care diagnostic tests, i.e. CM or histology, were performed locally. |
PMC12317575 | Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study | To investigate the feasibility of performing multi-modal bone marrow disease analysis on the same sample, we first evaluated how often each technique was successfully performed in our cohort (Table 2). |
PMC12317575 | Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study | Due to low cellularity in 96 samples (19%), no cytospins could be made and thus AIPF was performed on 413 samples. |
PMC12317575 | Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study | Twelve (3%) were not evaluable by AIPF due to high autofluorescence or low cell counts. |
PMC12317575 | Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study | Sequential iFISH analysis visualizing tumor-typical cytogenetic aberrations was performed in forty ambiguously positive samples. |
PMC12317575 | Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study | In 25/40 (63%), iFISH confirmed the same number of positive cells as with automated immunofluorescence alone. |
PMC12317575 | Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study | In 15/40 (38%), the number of positive cells detected by AIPF differed from that detected by automated immunofluorescence. |
PMC12317575 | Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study | However, only in 7/15 (47%), iFISH-result classified samples as negative or led to inconclusive results. |
PMC12317575 | Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study | Interestingly, 5 of these 7 (71%) were RT-qPCR positive. |
PMC12317575 | Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study | Of the 509 samples collected, neuroblastoma mRNA detection failed in nine (2%) because of low cDNA concentration. |
PMC12317575 | Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study | However, in eight of these cases, the paired bone marrow sample collected from the contralateral side yielded sufficient cDNA and could be included in the analyses (yielding 508). |
PMC12317575 | Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study | Standard-of-care diagnostics (CM or histology) was performed in 505 samples of which 1/505 (0.2%) was not evaluable by either, CM or histology (118 CM only, 2 histology only, 384 CM and histology) (Fig. 1B and Table 2). |
PMC12317575 | Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study | Fig. 1Study design, cohort and benchmarking against standard of care bone marrow assessment. |
PMC12317575 | Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study | A Outline of study cohort and analytical workflows for automated immunofluorescence plus fluorescence in situ hybridization (AIPF), RT-qPCR and cytomorphology/histology. |
PMC12317575 | Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study | B Consort diagram depicting sample inclusion and exclusion. |
PMC12317575 | Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study | C Contribution of AIPF, RT-qPCR (adrenergic (ADRN)-mRNA markers) and cytomorphology (CM)/histology. |
PMC12317575 | Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study | Venn diagram shows samples positive for at least one technology. |
PMC12317575 | Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study | Each circle represents positive results of one technique. |
PMC12317575 | Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study | N = 395 samples analyzed by all three techniques. |
PMC12317575 | Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study | N = 236 samples positive by ≥ 1 technique. |
PMC12317575 | Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study | D Representative microscopy image for GD2, CD56/NCAM and DAPI for a bone marrow specimen that was identified as positive only by AIPF, and negative by RT-qPCR and CM/histology. |
PMC12317575 | Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study | Scale bar represents 20 µm. |
PMC12317575 | Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study | E Level of tumor cell infiltration according to AIPF (y-axis; give n as % DTCs detected by AIPF) in specimens with single or combined positivity for AIPF, RT-qPCR and CM/histology (x-axis; + positive,—negative). |
PMC12317575 | Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study | Box plots represent 10–90 percentiles, line shows median. ** |
PMC12317575 | Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study | = 0.0023; *** = 0.0003. |
PMC12317575 | Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study | F Level of tumor cell infiltration according to RT-qPCR-ADRN (y-axis; given as % relative to neuroblastoma cell line IMR32) in specimen with single or combined positivity for AIPF, RT-qPCR-ADRN and CM/histology (x-axis; + positive,—negative). |
PMC12317575 | Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study | Box plots represent 10–90 percentiles, line shows median. * |
PMC12317575 | Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study | = 0.0135; ** = 0.0007; *** < 0.0001 Study design, cohort and benchmarking against standard of care bone marrow assessment. |
PMC12317575 | Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study | A Outline of study cohort and analytical workflows for automated immunofluorescence plus fluorescence in situ hybridization (AIPF), RT-qPCR and cytomorphology/histology. |
PMC12317575 | Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study | B Consort diagram depicting sample inclusion and exclusion. |
PMC12317575 | Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study | C Contribution of AIPF, RT-qPCR (adrenergic (ADRN)-mRNA markers) and cytomorphology (CM)/histology. |
PMC12317575 | Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study | Venn diagram shows samples positive for at least one technology. |
PMC12317575 | Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study | Each circle represents positive results of one technique. |
PMC12317575 | Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study | N = 395 samples analyzed by all three techniques. |
PMC12317575 | Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study | N = 236 samples positive by ≥ 1 technique. |
PMC12317575 | Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study | D Representative microscopy image for GD2, CD56/NCAM and DAPI for a bone marrow specimen that was identified as positive only by AIPF, and negative by RT-qPCR and CM/histology. |
PMC12317575 | Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study | Scale bar represents 20 µm. |
PMC12317575 | Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study | E Level of tumor cell infiltration according to AIPF (y-axis; give n as % DTCs detected by AIPF) in specimens with single or combined positivity for AIPF, RT-qPCR and CM/histology (x-axis; + positive,—negative). |
PMC12317575 | Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study | Box plots represent 10–90 percentiles, line shows median. ** |
PMC12317575 | Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study | = 0.0023; *** = 0.0003. |
PMC12317575 | Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study | F Level of tumor cell infiltration according to RT-qPCR-ADRN (y-axis; given as % relative to neuroblastoma cell line IMR32) in specimen with single or combined positivity for AIPF, RT-qPCR-ADRN and CM/histology (x-axis; + positive,—negative). |
PMC12317575 | Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study | Box plots represent 10–90 percentiles, line shows median. * |
PMC12317575 | Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study | = 0.0135; ** = 0.0007; *** < 0.0001 Table 1Patient characteristicsNumber of patients108Country Austria39 Netherlands69Stage (INSS) 32 4105 Unknown1Age at diagnosis (months) < 1825 ≥ 1883 Median36 Range3–274Sex Male65 Female43MYCN amplification status Amplified38 Non-amplified69 Unknown1INSS International Neuroblastoma Staging System Patient characteristics INSS International Neuroblastoma Staging System Table 2Technique performance on bone marrow liquid biopsy samples from patients with high-risk neuroblastomaSamples analyzed[number]Analysis successful[number]Analysis failed [number (%)]Samples not analyzed [number (%)]AIPF*41340112 (3%)96 (19%)Cell numbersiFISHperformed<1 million3229 131-2 million104104130≥2 million264264250Unknown134 19RT-qPCR*5095081 (0.2%)GUS Ct >25Not applicableOne side only: 8Both sides: 1CM*503435Suspicious: 268 (13.5%)Not representative: 36Inconclusive: 63Dubious: 2Histology*387333Suspicious: 1254 (14%)Not representative: 5122Inconclusive: 49 Total number of samples collected n 509 Technique performance on bone marrow liquid biopsy samples from patients with high-risk neuroblastoma iFISH performed Total number of samples collected n 509 Of the 413 samples exchanged, 184 (45%) were from Austrian and 229 (55%) from Dutch patients (Supplemental Fig. 1A). |
PMC12317575 | Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study | For > 95% of these, it was possible to perform AIPF and RT-qPCR, as well as cytomorphological examination of bone marrow smears and/or histological examination of bone trephines (Supplemental Fig. 1B). |
PMC12317575 | Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study | In total 395 samples were analyzed by AIPF, RT-qPCR and CM/histology (Fig. 1B). |
PMC12317575 | Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study | Sample preparation time (i.e. time from sampling to cytospin preparation), was shorter for Austrian bone marrow aspirates (Supplemental Fig. 1C). |
PMC12317575 | Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study | Mean cell counts were 3.80 million (range 130,000–7.97 million) for Austrian and 1.95 million (237,000–5.00 million) for Dutch samples (Supplemental Fig. 1D). |
PMC12317575 | Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study | The median turnaround time (sample collection to clinical report) for AIPF was 6 days (range 1 to 29 days), for the Austrian samples, where the results are used clinically. |
PMC12317575 | Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study | Two hundred thirty-six (236) bone marrows (60%) were positive for one or more techniques (Fig. 1B). |
PMC12317575 | Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study | We conclude that performing all three analyses on the same bone marrow specimen is feasible in an international multi-center setting. |
PMC12317575 | Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study | In the cohort of 395 multi-modal bone marrow analyses, we next determined the contribution of each technique to MRD detection and the link of DTC infiltration to uni- versus multi-modal assessment. |
PMC12317575 | Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study | In the 395 samples analyzed by all three techniques—AIPF, RT-qPCR-ADRN and CM/histology—64 (16%) were positive by all techniques and 159 (40%) negative. |
PMC12317575 | Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study | Among the 236 bone marrow specimens (60%) that showed positivity for at least one of the techniques, RT-qPCR-ADRN detected bone marrow disease more frequently than AIPF or CM/histology (216 RT-qPCR-ADRN vs. 108 AIPF vs. 112 CM/histology). |
PMC12317575 | Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study | Additionally, RT-qPCR-ADRN showed exclusive positivity in eighty, AIPF in four and CM/histology in sixteen cases (Fig. 1C & D & Supplemental Table 2). |
PMC12317575 | Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study | Interestingly, in CM/histology samples (n = 283), MRD-detection by AIPF was positive in 16% (n = 44) and RT-qPCR-ADRN was positive in 42% (n = 120); two of these cases were taken at diagnosis: both were RT-qPCR and one AIPF. |
Subsets and Splits
No community queries yet
The top public SQL queries from the community will appear here once available.