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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 Representative case (patient 166) showing MES marker positivity prior to relapse.
PMC12317575
Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study
At diagnosis adrenal tumor with MYCN amplification and multiple bone metastases and bone marrow infiltration; patient suffered a relapse one month after completion of SIOPEN/HR-NBL1 treatment and died 20 months later from disease.
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 samples assessed by RT-qPCR-ADRN and RT-qPCR-MES (normalized mRNA expression left y-axis) and AIPF (given as % DTCs, right y-axis) per timepoint (x-axis).
PMC12317575
Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study
DE = diagnosis, RE1 = mid-induction chemotherapy, RE2 = end of induction therapy, RE3 = surgery, RE4 = before stem cell transplantation, RE5 = before immunotherapy, RES6 = mid-immunotherapy, RE7 = at end of immunotherapy Mesenchymal markers identify MRD in adrenergic negative and AIPF negative bone marrow liquid biopsies prior to relapse.
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 Contribution of mesenchymal mRNA RT-qPCR-markers (RT-qPCR-MES),RT-qPCR-ADRN, AIPF 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 techniques; N = 273 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
B MES-positive samples by RT-qPCR-MES.
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 infiltration according to AIPF (y-axis; given as % DTCs) and RT-qPCR-ARDN (x-axis; given as % relative to neuroblastoma cell lines IMR32) (n = 107; Spearman correlation = 0.59, 95% CI 0.33–0.77; p < 0.001 of samples positive for both techniques (n = 38).
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 Level of infiltration by AIPF (y-axis; given as % DTCs) versus MES-positivity (x-axis).
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
Heatmap represents number of negative and positive cases. *
PMC12317575
Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study
= 0.0377.
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 Level of infiltration by RT-qPCR-ADRN (y-axis; given as % relative to neuroblastoma cell line IMR32) versus MES-positivity (x-axis).
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
Heatmap represents number of negative and positive cases. **
PMC12317575
Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study
= 0.0029.
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 Representative case (patient 166) showing MES marker positivity prior to relapse.
PMC12317575
Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study
At diagnosis adrenal tumor with MYCN amplification and multiple bone metastases and bone marrow infiltration; patient suffered a relapse one month after completion of SIOPEN/HR-NBL1 treatment and died 20 months later from disease.
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 samples assessed by RT-qPCR-ADRN and RT-qPCR-MES (normalized mRNA expression left y-axis) and AIPF (given as % DTCs, right y-axis) per timepoint (x-axis).
PMC12317575
Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study
DE = diagnosis, RE1 = mid-induction chemotherapy, RE2 = end of induction therapy, RE3 = surgery, RE4 = before stem cell transplantation, RE5 = before immunotherapy, RES6 = mid-immunotherapy, RE7 = at end of immunotherapy When considering only discordant pairs, the number of samples positive for the combination but CM/histology was 10.7 times higher (95% CI, 6.3–18.2) than the number of samples positive by CM/histology alone (Table 3), demonstrating that the combined test is superior to the current standard-of-care, 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
Of the 37 samples exclusively positive for MES mRNAs and negative by the other techniques, fourteen cases were from Dutch patients for which 12/14 plasma samples were available.
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 all twelve samples we detected hypermethylated RASSF1A DNA, a marker for neuroblastoma for which our group previously designed a droplet digital PCR assay .
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 further investigated eight samples from patients carrying a MYCN amplification in tumor cells which were RT-qPCR-MESAIPF (five RT-qPCR-ADRN, three -ADRN); however, using MYCN iFISH, no cells with MYCN amplification were detectable.
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 38 cases positive for AIPF, RT-qPCR-MES and -ADRN, tumor cell infiltration covered a range of 1 DTC in 5.30 million to 2.23 million DTCs in 2.48 million MNCs according to AIPF and 0.0001–341% (relative to IMR32) according to RT-qPCR, and correlated well (Fig. 4B).
PMC12317575
Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study
There was a slightly higher tumor cell infiltration detectable in bone marrow-positive compared to those negative for MES mRNAs, as determined by AIPF (the number of investigated MNCs did not differ between MES and MES) and RT-qPCR-ADRN (Fig. 4C & D and Supplemental Fig. 4C).
PMC12317575
Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study
This indicates that RT-qPCR-MES shows lower sensitivity, yet might identify MRD in an additional set of 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
Interestingly, in patients with serially collected samples along multi-modal treatment and follow-up, signals often alternated between different 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
This is exemplified by patient 166 and others, who showed an initial increase of MES mRNAs and delayed clearance upon induction chemotherapy, with an early increase of MES mRNA POSTN prior to relapse (Fig. 4E and Supplemental Fig. 5A-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
Sensitive bone marrow MRD detection is crucial for evaluating therapy response, monitoring disease progression, and early relapse detection, helping to identify patients who may benefit from alternative treatments.
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, assessing tumor cell state plasticity and the abundance of immunotherapy targets is essential for accurately evaluating immunotherapy efficacy.
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 prospective study, we address these needs using multi-modal, high-sensitivity MRD techniques in 509 samples obtained from 108 patients.
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 show that (1) performing all three high-sensitivity analyses, i.e. AIPF, RT-qPCR-ADRN and -MES on the same bone marrow specimen, alongside standard-of-care assessments, is feasible in an international multicenter setting; (2) multi-modal analysis is more effective at detecting bone marrow MRD than the current standard of care, with low tumor cell burden detected by RT-qPCR-ADRN more frequently and less frequently by AIPF; (3) bone marrow disease is present on both sides when DTC infiltration is high (e.g., at diagnosis).
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 MRD settings during and post-treatment, more frequently only one side is affected; (4) AIPF and RT-qPCR are complementary approaches for high-sensitivity MRD detection and characterizing of tumor cell phenotypes and therapeutic targets; and (5) RT-qPCR-MES shows lower sensitivity but may identify MRD in an additional subset of 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
Notably, in a patient with initially high MES markers, these markers showed delayed clearance post-induction chemotherapy and an early increase before relapse.
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 have previously demonstrated the superior sensitivity of mRNA- and DTC-based bone marrow MRD detection 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
RT-qPCR-based testing of bone marrow and peripheral blood neuroblastoma patients is a robust and highly sensitive method for detecting residual disease [19, 31–34, 36–40, 54, 55].
PMC12317575
Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study
Stutterheim, et al. reported RT-qPCR positivity in 31% (5/16) of bone marrow GD2-immunocytology samples and Hartomo, et al. reported RT-qPCR positivity in 48% (30/63) of bone marrow-cytology 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
We have further shown in 345 patients that molecular detection of MRD by RT-qPCR was more sensitive and had higher prognostic value than GD2-immunocytology for neuroblastoma BM infiltration .
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 contrast, AIPF can detect up to one tumor cell per million cells and provides a more automated and objective alternative to manual CM or immunocytology assessment based solely on GD2 .
PMC12317575
Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study
AIPF has been compared against flow cytometry and cytomorphology in a study by Schriegel, et al. involving 369 bone marrow aspirates .
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 latter study, AIPF was demonstrated to be the single most sensitive method for the detection of bone marrow involvement in neuroblastoma .
PMC12317575
Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study
Furthermore, at end of induction chemotherapy, DTC-positivity by AIPF was significantly associated with event-free survival and cumulative incidence of relapse in a study of 180 patients (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
In the prospective study described here, we combined these complementary techniques, i.e. RT-qPCR-ADRN, AIPF and RT-qPCR-MES, to evaluate their effectiveness in detecting MRD across 509 bone marrow aspirates from 108 patients in a multicenter 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
Our findings indicate that multicenter exchange is feasible within a clinically relevant six-day turnaround time.
PMC12317575
Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study
Despite differences in treatment protocols (NCT01704716, NCT01704716) and cut-offs for CM assessment , and differences in mean cell counts (3.80 million (range 130,000–7.97 million) for Austrian and 1.95 million (range 237,000–5.00 million) for Dutch samples) no major differences in results were observed between Austrian and Dutch samples compared to aggregated 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
This underlines the ability to perform high-sensitivity analyses from small bone marrow volumes in an international setting and demonstrates the robustness of the techniques, paving the way for standardized MRD detection across clinical settings.
PMC12317575
Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study
This standardization is important for the ongoing European SIOPEN High-Risk Neuroblastoma 2 (HR-NBL2) trial (NCT04221035), ensuring accurate and timely diagnosis.
PMC12317575
Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study
Moreover, in our study, RT-qPCR-ADRN emerges as the most sensitive method for MRD detection.
PMC12317575
Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study
It is important to emphasize that AIPF uniquely identified certain samples not detected by RT-qPCR, suggesting that AIPF provides complementary information by enabling direct visualization and genetic verification of disseminated tumor cells, thereby offering insights into tumor cell morphology and immunotherapy marker expression that molecular methods alone cannot provide.
PMC12317575
Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study
As AIPF quantifies actual cells while RT-qPCR measures transcripts, a strong correlation between the techniques is evident when tumor cell infiltration is high, i.e., at diagnosis and at an event.
PMC12317575
Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study
Our findings confirm previous observations that PHOX2B sensitivity is most evident in samples with low tumor infiltration, such as bone marrow samples during treatment .
PMC12317575
Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study
Notably, bone marrow infiltration levels detected by AIPF and RT-qPCR decreased during treatment and increased at relapse, reinforcing the potential of these methods response monitoring and early relapse, as well as highlighting the importance of serial sampling.
PMC12317575
Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study
Future studies should explore their association in treatment response as predictive value for survival.
PMC12317575
Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study
While our study did not recommend a reduction of bilateral BM sampling, the sensitivity of AIPF and RT-qPCR suggests unilateral sampling at diagnosis may suffice for initial bone marrow disease 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
This could reduce patient burden and costs, with bilateral sampling reserved for follow-up when tumor cell infiltration is lower.
PMC12317575
Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study
Future studies should explore this approach.
PMC12317575
Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study
Our results also facilitate implementation as clinical decision-making tools for detecting molecular relapse, crucial for timely intervention.
PMC12317575
Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study
Prospective trials are currently underway to validate the use of RT-qPCR-ADRN as a diagnostic tool in blood liquid biopsies to determine imminent molecular relapse early.
PMC12317575
Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study
Beyond MRD detection, understanding therapy suitability is essential.
PMC12317575
Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study
Recent studies highlight distinct vulnerabilities of MES and ADRN neuroblastoma phenotypes.
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 cells exhibit in vitro, more resistant to chemotherapeutics, retinoic acid, and ALK inhibition , drugs that are applied in current first- and second-line treatment protocols for patients with neuroblastoma .
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 contrast, NOTCH pathway downregulation adversely affects MES cell survival in vitro and in vivo .
PMC12317575
Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study
Our study shows that MES and ADRN phenotypes fluctuate during therapy, with MES-related mRNAs increasing before relapse in a patient with metastatic disease and slow induction therapy response.
PMC12317575
Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study
This corroborates previous work by our group in a retrospective cohort (95 bone marrow samples from 38 patients), where MES mRNA-expression increased in 18/27 patients during induction, while ADRN mRNA-expression initially decreased but was elevated at relapse in 9/12 patients .
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, MES-related mRNAs were detected in the absence of detectable ADRN mRNAs or DTCs in patients who did not relapse.
PMC12317575
Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study
Future research should determine whether these findings indicate MES tumor cells or other mesenchymal-like processes are triggered or induced by therapy or fibrosis which can result in tumor cell senescence .
PMC12317575
Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study
Monitoring the appearance of MES cells can provide insights into treatment response and potential resistance.
PMC12317575
Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study
Tools that accurately detect and characterize these phenotypes are useful for tailoring treatment plans to individual patients.
PMC12317575
Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study
Therapies targeting MES-neuroblastoma may benefit from such monitoring, although wider prospective studies are needed to validate these findings.
PMC12317575
Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study
Anti-GD2 immunotherapy is a critical treatment for high-risk neuroblastoma and other cancers .
PMC12317575
Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study
Our study shows that GD2 and other immunotherapy targets can be monitored and quantified using AIPF and advanced image-based multiplexed approaches.
PMC12317575
Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study
Anti-GD2 therapy, after successful trials in relapsed/refractory patients , is transitioning to frontline treatment as part of chemo-immunotherapy.
PMC12317575
Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study
Flow cytometry has been successfully used in 41 bone marrow specimens from 25 patients to determine GD2 levels on DTCs at diagnosis and relapse when tumor cells are abundant .
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 demonstrate that GD2 can be detected even when tumor cells are extremely rare, e.g. during treatment.
PMC12317575
Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study
Measuring GD2 and other immunotherapy targets (e.g., NCAM, L1CAM, GPC-2) in therapy-resistant cells is essential for assessing treatment efficacy and making informed clinical decisions.
PMC12317575
Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study
AIPF, as one of the few ISO-certified tests for GD2 detection, will be integral in multi-modal diagnostic workflows as antibody and CAR-T cell-based immunotherapies gain traction in neuroblastoma [25, 26, 28, 66–69] and other cancers, such as Ewing sarcoma and melanoma , highlighting the broader applicability of these monitoring 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
Limitations of this study include the following aspects: First, this study is well-powered for technical comparison and proof-of-principle demonstration, however, clinical relevance of the combined test in detecting treatment response, early relapse, or evaluating its predictive or prognostic power cannot be determined.
PMC12317575
Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study
Its clinical relevance is currently being assessed prospectively in the SIOPEN frontline trial HR-NBL2 (NCT04221035) and other trials enrolling patients with high-risk neuroblastoma.
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 second limitation is the current clinical practice for collecting bone marrow aspirates, which mandates using the first drop of aspirates for smear preparation to assess cytomorphology .
PMC12317575
Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study
Consequently, bone marrow samples used for RT-qPCR and AIPF are more diluted with blood, potentially explaining why sixteen samples were positive by CM/histology only.
PMC12317575
Sensitive detection of minimal residual disease and immunotherapy targets by multi-modal bone marrow analysis in high-risk neuroblastoma – a multi-center study
Lastly, AIPF is highly robust in detecting GD2 and CD56 on tumor cells, particularly when combined with iFISH, which includes adequate positive and negative controls in each analysis run.
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, detecting tumor cells negative for both markers remains challenging and will require techniques with carefully designed, highly multiplexed marker panels, such as imaging mass cytometry or flow cytometry.
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 clinical application it will be important to carefully design such marker panels and develop in future prospective studies cost-effective lab tests for highly sensitive methods such as RT-qPCR, AIPF, and multiparameter flow cytometry.
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 conclusion, our study highlights the importance of advanced multi-modal MRD detection techniques, monitoring tumor phenotype switches, and assessing immunotherapy targets to improve patient outcomes in neuroblastoma and other cancers.
PMC12602814
LPS-Induced Neuroinflammation Disrupts Brain-Derived Neurotrophic Factor and Kinase Pathways in Alzheimer's Disease Cell Models
Alzheimer's disease (AD) leads to a progressive loss of cognitive abilities and memory.
PMC12602814
LPS-Induced Neuroinflammation Disrupts Brain-Derived Neurotrophic Factor and Kinase Pathways in Alzheimer's Disease Cell Models
A critical factor now recognized as driving AD pathology is neuroinflammation—inflammation occurring in the nervous system, which contributes to neuronal harm and communication breakdowns.
PMC12602814
LPS-Induced Neuroinflammation Disrupts Brain-Derived Neurotrophic Factor and Kinase Pathways in Alzheimer's Disease Cell Models
our research investigated the specific effects of neuroinflammation on neuronal signaling pathways.
PMC12602814
LPS-Induced Neuroinflammation Disrupts Brain-Derived Neurotrophic Factor and Kinase Pathways in Alzheimer's Disease Cell Models
In this study, we primarily employed the SH-SY5Y neuroblastoma cell line as an in vitro neuronal model to investigate inflammatory responses relevant to AD etiology, alongside supplementary observations in primary neurons and 3D spheroids for comparative analysis.
PMC12602814
LPS-Induced Neuroinflammation Disrupts Brain-Derived Neurotrophic Factor and Kinase Pathways in Alzheimer's Disease Cell Models
Our analysis focused on modifications of key molecules, including the neuroprotective protein Brain-Derived Neurotrophic Factor (BDNF), pro-inflammatory cytokines such as IL-6 and TNF-α, and crucial regulatory kinases.
PMC12602814
LPS-Induced Neuroinflammation Disrupts Brain-Derived Neurotrophic Factor and Kinase Pathways in Alzheimer's Disease Cell Models
Our results demonstrated that LPS treatment dramatically lowered the vitality and decreased BDNF levels in the SH-SY5Y cells.
PMC12602814
LPS-Induced Neuroinflammation Disrupts Brain-Derived Neurotrophic Factor and Kinase Pathways in Alzheimer's Disease Cell Models
Furthermore, we observed a considerable elevation in the pro-inflammatory cytokines IL-6 and TNF-α, coupled with elevated levels of COX-2 and iNOS.
PMC12602814
LPS-Induced Neuroinflammation Disrupts Brain-Derived Neurotrophic Factor and Kinase Pathways in Alzheimer's Disease Cell Models
Gene expression data validated that LPS treatment altered the expression of essential signaling kinases (Protein Kinase A (PKA), Protein Kinase B (AKT), and Mitogen-Activated Protein Kinase (MAPK)).
PMC12602814
LPS-Induced Neuroinflammation Disrupts Brain-Derived Neurotrophic Factor and Kinase Pathways in Alzheimer's Disease Cell Models
Our first comparative analysis revealed that 3D spheroid cultures may elicit more pronounced inflammatory responses than standard 2D cultures; nevertheless, our detailed investigation primarily focused on the SH-SY5Y model.
PMC12602814
LPS-Induced Neuroinflammation Disrupts Brain-Derived Neurotrophic Factor and Kinase Pathways in Alzheimer's Disease Cell Models
This study revealed that LPS-induced neuroinflammation affects neuronal signaling in vitro, thereby revealing a relationship between inflammation and neuronal dysfunction in cellular models of neuroinflammation.
PMC12602814
LPS-Induced Neuroinflammation Disrupts Brain-Derived Neurotrophic Factor and Kinase Pathways in Alzheimer's Disease Cell Models
These findings highlight pathways that may be relevant to AD pathophysiology; however, further in vivo studies are necessary to demonstrate their translational relevance to humans.
PMC12602814
LPS-Induced Neuroinflammation Disrupts Brain-Derived Neurotrophic Factor and Kinase Pathways in Alzheimer's Disease Cell Models
This graphical abstract illustrates how LPS-induced neuroinflammation disrupts neurotrophic signaling and kinase activity in Alzheimer's models.
PMC12602814
LPS-Induced Neuroinflammation Disrupts Brain-Derived Neurotrophic Factor and Kinase Pathways in Alzheimer's Disease Cell Models
It highlights the involvement of SH-SY5Y cells, primary neurons, and 3D spheroids in inflammatory cascades that lead to oxidative stress, BDNF suppression, and synaptic dysfunction, offering insights into potential therapeutic targets.
PMC12602814
LPS-Induced Neuroinflammation Disrupts Brain-Derived Neurotrophic Factor and Kinase Pathways in Alzheimer's Disease Cell Models
AD is a progressive neurodegenerative disorder characterized by cognitive decline, memory loss, and synaptic malfunction.