Building a B Cell Differentiation Model for X-Linked Agammaglobulinemia Using Pluripotent Stem Cells
Abstract/Contents
- Abstract
- X-Linked Agammaglobulinemia (XLA) is the most common primary immunodeficiency in males in the United States. It is characterized by the inability to produce mature B cells due to mutations in the BTK gene. Current treatments involve repeated injection of immunoglobulins, but these have limitations, as patients can have adverse reactions and this treatment does not replace the entire immune response. As a result, gene correction is considered a viable therapy for XLA; however, a robust B cell differentiation model to study the effect of using gene therapy to treat XLA has not been established. Although cytokines are not sufficient to induce differentiation, studies have shown that transcription factor-driven differentiation systems have been proven to be successful in producing differentiated cells from pluripotent stem cells (PSCs). Therefore, this project aimed to study and replicate B cell development in vitro with the purpose of designing a new differentiation model using B cell-specific transcription factors (TFs), EBF1 and PAX5. This was approached by using the gene editing tool clustered regularly interspaced short palindromic repeats-Cas9 (CRISPR-Cas9) and adeno-associated virus type 6 (AAV6) on human PSCs. First, assessment of wild type PSC potential to differentiate into B cells using cytokines confirmed inefficient B cell development. Then, an in-depth study of BTK mutations led to the generation of three pluripotent XLA-knockout cell lines that imitate the disease phenotype. The design and integration of GFP reporter constructs into the B-cell specific marker CD20 is still being studied. Finally, TF constructs were designed with the purpose of enhancing a cytokine-based B cell differentiation. These results support the development of PSC lines with transgenic B cell differentiation TFs as a model for XLA and other B cell-related malignancies, while eliminating the need for patients to provide bone marrow or hematopoietic stem cell samples.
Description
| Type of resource | text |
|---|---|
| Publication date | May 5, 2023 |
Creators/Contributors
| Author | Hernández González, Elaine |
|---|---|
| Thesis advisor | Porteus, Matthew |
| Thesis advisor | Ghanim, Hana |
| Thesis advisor | Dixon, Scott |
| Degree granting institution | Stanford University, Department of Biology |
Subjects
| Subject | B cell |
|---|---|
| Subject | Differentiation |
| Subject | Transcription factors |
| Subject | Pluripotent stem cells |
| Subject | X-Linked Agammaglobulinemia |
| Genre | Text |
| Genre | Thesis |
Bibliographic information
Access conditions
- License
- This work is licensed under a Creative Commons Attribution Non Commercial 4.0 International license (CC BY-NC).
Preferred citation
- Preferred citation
- Hernández González, E. and Porteus, M. (2024). Building a B Cell Differentiation Model for X-Linked Agammaglobulinemia Using Pluripotent Stem Cells. Stanford Digital Repository. Available at https://purl.stanford.edu/nf436hf9370. https://doi.org/10.25740/nf436hf9370.
Collection
Undergraduate Theses, Department of Biology, 2022-2023
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- Contact
- elainehg@stanford.edu
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