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A Multi-Color Flow Cytometric Assay for Quantifying Dinutuximab Binding to Neuroblastoma Cells in Tumor, Bone Marrow,
Michelle E Keyel1, Kathryn L Furr1, Min H Kang2
1Department of Pediatrics, School of Medicine, Texas Tech University Health Sciences Center, Lubbock, TX 79430, USA.
Abstract:
GD2, a disialoganglioside, is present on the surface of most neuroblastomas, as well as on some other cancers, such as melanoma and osteogenic sarcoma. The anti-GD2 antibody ch14.18 (dinutuximab) has an FDA-registered indication for use as maintenance therapy for high-risk neuroblastoma with cytokines and 13-cis-retinoic acid after myeloablative therapy. Recent studies using immunohistochemistry of tumor or tumor cells in marrow have shown that some neuroblastomas are negative for GD2. Dinutuximab and other anti-GD2 antibodies are increasingly used in combination with cytotoxic chemotherapy for treating relapsed neuroblastoma, so it is important to be able to identify patients with tumor cells with low GD2 expression, as such patients may experience toxicity but not benefit from the antibody therapy. As the most common clinical samples available for relapsed neuroblastoma are bone marrow aspirates, we developed a method to quantify dinutuximab binding density and the frequency of neuroblastoma cells positive for the antibody in bone marrow aspirates. Here, we describe a multi-color flow cytometry assay that employs non-GD2 antibodies to identify neuroblastoma cells in a mixed population (tumor, bone marrow, or blood) and an anti-GD2 antibody to quantify both the frequency and density of GD2 expression on neuroblastoma cells.
Insights
A new flow cytometry assay quantifies GD2 expression on neuroblastoma cells in bone marrow. This method helps identify patients unlikely to benefit from anti-GD2 antibody therapy, optimizing treatment for relapsed neuroblastoma.
Area of Science:
- Oncology
- Immunology
- Biotechnology
Background:
- GD2 is a key target antigen on neuroblastoma cells.
- The anti-GD2 antibody dinutuximab is FDA-approved for high-risk neuroblastoma maintenance therapy.
- Some neuroblastomas exhibit low or negative GD2 expression, potentially limiting treatment efficacy and increasing toxicity.
Purpose of the Study:
- To develop a method for quantifying GD2 expression on neuroblastoma cells in bone marrow aspirates.
- To identify patients with low GD2 expression who may not benefit from anti-GD2 antibody therapy.
- To improve risk stratification and treatment selection for relapsed neuroblastoma.
Main Methods:
- Development of a multi-color flow cytometry assay.
- Utilized non-GD2 antibodies to identify neuroblastoma cells within mixed populations (bone marrow, blood).
- Quantified dinutuximab binding density and frequency of GD2-positive cells using an anti-GD2 antibody.
Main Results:
- Successfully developed and validated a flow cytometry assay for GD2 quantification in bone marrow.
- The assay can determine both the frequency and density of GD2 expression on neuroblastoma cells.
- This method allows for the identification of neuroblastoma patients with low GD2 expression.
Conclusions:
- The described flow cytometry assay is a valuable tool for assessing GD2 expression in relapsed neuroblastoma.
- Accurate GD2 quantification can guide therapeutic decisions, potentially avoiding ineffective treatments and associated toxicities.
- This assay supports personalized medicine approaches in neuroblastoma treatment.
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