Generating an in-vitro Model of MPS-IIIA “Sanfilippo Syndrome” via CRISPR/CAS9 and Exploring Potential Therapeutics in a PUI Setting
Document Type
Poster Presentation
Publication Date
4-25-2025
Keywords
fsc2025
Abstract
Autosomal recessive inheritance of a defective SGSH gene coding for the enzyme sulfamidase is responsible for mucopolysaccharidosis (MPS) type IIIA, characterized by the effects of heparan sulfate accumulation in lysosomes. The syndrome presents with coarse facial features, central nervous system dysfunction, behavioral difficulties, intellectual disability, sleep disturbance, hyperactivity, loss of communication skills, and loss of mobility. Failure to develop cost effective research methods and long-term treatment options leaves the average age expectancy at just 10-20 years of age. Gene editing techniques including ZFNs, TALENs, and CRISPR/Cas9 have been explored as both models and potential therapeutic interventions for mucopolysaccharidosis types I, II and III. However, an in-vitro model of MPS IIIA, the most severe type of MPS, has yet to be developed. Therefore, we present a model test an in-vitro human cell line model of MPS IIA using a K562 lymphoblast cell line, CRISPR/Cas9 gene editing, and a heparan sulfate enzyme-linked immunosorbent assay (ELISA). We hypothesized that disruption of SGSH would reduce or eliminate function of the sulfamidase enzyme, resulting in toxic heparan sulfate accumulation in the cell that could be quantified by an ELISA. Furthermore, we hypothesized Fluoxetine, an experimental drug treatment, would reduce heparan sulfate levels in the cell based on previous evidence in an MPS-IIIA murine model. Our work demonstrated the K562 lymphoblast cell line is a cost-effective and viable cell type for quantifying heparan sulfate levels. Preliminary drug treatments also suggest Fluoxetine may be capable of reducing cellular heparan sulfate levels in-vitro. Altogether, this work points to the potential of creating a sustainable cell model of MPS-IIIA, with the possibility of recapitulating specific patient mutations via the CRISPR/Cas9 editing system in the future.
Publication Information
Cooper, Emma and Murphy, Zachary, "Generating an in-vitro Model of MPS-IIIA “Sanfilippo Syndrome” via CRISPR/CAS9 and Exploring Potential Therapeutics in a PUI Setting" (2025). Fisher Showcase 2025. Paper 32.
https://fisherpub.sjf.edu/fsc2025/32
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Comments
Student poster presented at the 2025 Fisher Showcase, St. John Fisher University, April 25, 2025.
Also presented at NCUR 25, Pittsburgh, PA, April 7-9, 2025