Internalization of anti-GD2 antibodies as a key component of the antibody-induced cell death mechanism in pediatric neuroblastoma
Abstract/Contents
- Abstract
- Neuroblastoma is one of the most common solid cancers diagnosed in young children. Despite advancements in modern therapies, many high-risk patients experience recurrence and eventually die of their disease. Currently, front-line treatment therapy includes treatment with monoclonal antibodies recognizing the disialoganglioside GD2, a type of cell surface lipid. Neuroblastoma is characterized by the presence of GD2 on cancer cells, but recent data has shown that GD2 levels can be heterogeneous. In addition, other gangliosides such as GD3 and GM2 are present at varying levels and may serve as potential targets. It has been shown in the literature that anti-GD2 antibodies are able to cause cell death in an immune independent way that differs from classical antibody-induced cell death pathways such as antibody-dependent cell-mediated cytotoxicity and antibody-dependent cellular phagocytosis, though no consensus regarding the specific mechanism has been reached. This research aims to identify key genes in this cell death pathway and their effects on neuroblastoma and ganglioside levels. A CRISPR screen for genes that protect neuroblastoma cells from death due to anti-ganglioside antibodies yielded the candidate genes AP2M1 and AP2S1. The genes were knocked out to create cell lines for cell killing assays and assessed for protection against the anti-GD2 antibody dinutuximab. The levels of GD2 and other gangliosides in cell lines were monitored via flow cytometry. Cell-killing assays using dinutuximab showed decreased cell death in AP2M1 and AP2S1 knockout lines. Since AP2M1 and AP2S1 are genes involved in clathrin-mediated endocytosis, antibody internalization was monitored via time-lapse fluorescence microscopy for disruptions in the knockout cell lines. Internalization assays suggested that antibody internalization was necessary but not sufficient for neuroblastoma cell death. Furthermore, increased cell death and internalization was achieved in wildtype neuroblastoma cells when anti-GD2 antibodies were crosslinked. Altogether, these results provide more insight into antibody-ganglioside interactions in neuroblastoma and the internal cellular events that occur after antibody binding. There is great therapeutic potential in targeting internalization, offering a novel treatment applicable not only to pediatric neuroblastoma but also potentially to other types of cancers with similar ganglioside expression levels.
Description
| Type of resource | text |
|---|---|
| Publication date | May 4, 2023 |
Creators/Contributors
| Author | Wang, Alice |
|---|---|
| Thesis advisor | Majzner, Robbie |
| Thesis advisor | Dixon, Scott |
| Thesis advisor | Long, Sharon |
| Degree granting institution | Stanford University, Department of Biology |
Subjects
| Subject | Neuroblastoma |
|---|---|
| Subject | GD2 |
| Subject | Gangliosides |
| Subject | Monoclonal antibodies |
| Subject | dinutuximab |
| Subject | Cell death |
| Subject | internalization |
| Subject | AP2M1 |
| Subject | AP2S1 |
| Subject | clathrin-mediated endocytosis |
| Genre | Text |
| Genre | Thesis |
Bibliographic information
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- This work is licensed under a Creative Commons Attribution Non Commercial 4.0 International license (CC BY-NC).
Preferred citation
- Preferred citation
- Wang, A. and Majzner, R. (2023). Internalization of anti-GD2 antibodies as a key component of the antibody-induced cell death mechanism in pediatric neuroblastoma . Stanford Digital Repository. Available at https://purl.stanford.edu/dt072cf2338. https://doi.org/10.25740/dt072cf2338.
Collection
Undergraduate Theses, Department of Biology, 2022-2023
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- alice27@stanford.edu
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