Effects of Altered TPP-1 Expression in Human Neural Progenitor Cells
Abstract
Batten’s Disease or neuronal ceroid lipofuscinosis type 2 (CLN2) is a pediatric autosomal recessive, neurodegenerative lysosomal storage disorder due to a deficit of the lysosomal protease tripeptidyl peptidase (TPP-1). The lack of TPP-1 leads to an accumulation of lysosomal waste and cell death. CLN2 is characterized by language delays, seizures, cognitive and motor decline, blindness, and early death. Currently, a clinical trial of the experimental treatment Brineura® (Cerilponase Alfa) is the only approved treatment for CLN2. The clinical trial involves infusion of human recombinant TPP-1 (rhTPP1) into the ventricles of the brain. Diffusion models suggest the protein will spread along a concentration gradient through the brain. However, it is unclear how altered TPP-1 concentrations will affect ongoing neurogenesis in the subventricular zone. This study aims to examine proliferation, cell cycle kinetics, differentiation, and cell death in cultured human neural progenitor cells (hNPCs) following rhTPP-1 exposure. Altered levels of TPP-1 will be assessed using western blot analysis. Proliferation and differentiation were assessed using immunostaining, fluorescent microscopy, and CellProfiler™ analysis. Preliminary data on cell cycle kinetics was assessed using EdU-incorporation assays and suggest no changes in cell proliferation. Induced cell death indicated by apoptosis was assessed using ApopTag (TUNEL) assay. Results demonstrate increased levels of rhTPP-1 induced a significantly higher rate of cell death in healthy hNPCs; revealed through a one-way ANOVA: F (4,34) = 6.87, p = .0003, attributing increased protein concentration to increased cell death. These results may have implications in clinical trials using intracerebroventricular infusion for enzyme replacement therapies.
Faculty Sponsors
Dr. James Munoz
Project Type
Event
Location
Alvin Sherman Library
Start Date
4-3-2024 12:30 PM
End Date
4-4-2024 1:30 PM
Effects of Altered TPP-1 Expression in Human Neural Progenitor Cells
Alvin Sherman Library
Batten’s Disease or neuronal ceroid lipofuscinosis type 2 (CLN2) is a pediatric autosomal recessive, neurodegenerative lysosomal storage disorder due to a deficit of the lysosomal protease tripeptidyl peptidase (TPP-1). The lack of TPP-1 leads to an accumulation of lysosomal waste and cell death. CLN2 is characterized by language delays, seizures, cognitive and motor decline, blindness, and early death. Currently, a clinical trial of the experimental treatment Brineura® (Cerilponase Alfa) is the only approved treatment for CLN2. The clinical trial involves infusion of human recombinant TPP-1 (rhTPP1) into the ventricles of the brain. Diffusion models suggest the protein will spread along a concentration gradient through the brain. However, it is unclear how altered TPP-1 concentrations will affect ongoing neurogenesis in the subventricular zone. This study aims to examine proliferation, cell cycle kinetics, differentiation, and cell death in cultured human neural progenitor cells (hNPCs) following rhTPP-1 exposure. Altered levels of TPP-1 will be assessed using western blot analysis. Proliferation and differentiation were assessed using immunostaining, fluorescent microscopy, and CellProfiler™ analysis. Preliminary data on cell cycle kinetics was assessed using EdU-incorporation assays and suggest no changes in cell proliferation. Induced cell death indicated by apoptosis was assessed using ApopTag (TUNEL) assay. Results demonstrate increased levels of rhTPP-1 induced a significantly higher rate of cell death in healthy hNPCs; revealed through a one-way ANOVA: F (4,34) = 6.87, p = .0003, attributing increased protein concentration to increased cell death. These results may have implications in clinical trials using intracerebroventricular infusion for enzyme replacement therapies.
