Title

Novel Delivery of Mtor Inhibitor to Glioblastoma Cells

School Name

South Carolina Governor's School for Science & Mathematics

Grade Level

12th Grade

Presentation Topic

Cell and Molecular Biology

Presentation Type

Mentored

Abstract

Glioblastoma Multiforme (GBM) is an incurable brain cancer. Limited chemotherapeutic options are available to treat GBM. mTOR inhibitors could reduce GBM growth. Our research shows that GBM cells treated with Rapamycin, an mTOR inhibitor, experienced increased cell cytotoxicity as measured by an MTT cell viability assay. The brain cancer cells were plated in triplicate in 96-well plates and treated with increasing concentrations of free Rapamycin and encapsulated Rapamycin for 24 or 48 hours. MTT reagent, tetrazolium dye, was added to the wells and incubated for 2 hours. UV-VIS spectrophotometry was conducted. Rapamycin killed cells within 24 hours. Without retreatment, the cells recovered within 48 hours. Rapamycin must be in micelles because the blood brain barrier does not let mTOR inhibitors collect in the brain tumors. Micelles were made with PHC, PEG-PE, and Rapamycin, targeted with cRGD peptide and labeled with Dylight fluorophore. Micelle quality was characterized by UV-VIS and DLS. An uptake assay was conducted to quantify micelle uptake in GBM cells. Rapamycin encapsulated micelles (RaM) and targeted Rapamycin encapsulated micelles (TRaM) can be encapsulated in micelles with a diameter of 10-15 nm. Targeting integrins on the surface of GBM cells does not appear to improve uptake of the micelles compared to untargeted micelles. TRaM appears to reduce cell viability (~20%) within 24 hours. However, cells require retreatment daily to maintain cell cytotoxicity. Micelles do have an effect on cell viability and may lead to a more concentrated and localized treated of GBM cells.

Location

Furman Hall 107

Start Date

3-28-2020 10:45 AM

Presentation Format

Oral Only

Group Project

No

COinS
 
Mar 28th, 10:45 AM

Novel Delivery of Mtor Inhibitor to Glioblastoma Cells

Furman Hall 107

Glioblastoma Multiforme (GBM) is an incurable brain cancer. Limited chemotherapeutic options are available to treat GBM. mTOR inhibitors could reduce GBM growth. Our research shows that GBM cells treated with Rapamycin, an mTOR inhibitor, experienced increased cell cytotoxicity as measured by an MTT cell viability assay. The brain cancer cells were plated in triplicate in 96-well plates and treated with increasing concentrations of free Rapamycin and encapsulated Rapamycin for 24 or 48 hours. MTT reagent, tetrazolium dye, was added to the wells and incubated for 2 hours. UV-VIS spectrophotometry was conducted. Rapamycin killed cells within 24 hours. Without retreatment, the cells recovered within 48 hours. Rapamycin must be in micelles because the blood brain barrier does not let mTOR inhibitors collect in the brain tumors. Micelles were made with PHC, PEG-PE, and Rapamycin, targeted with cRGD peptide and labeled with Dylight fluorophore. Micelle quality was characterized by UV-VIS and DLS. An uptake assay was conducted to quantify micelle uptake in GBM cells. Rapamycin encapsulated micelles (RaM) and targeted Rapamycin encapsulated micelles (TRaM) can be encapsulated in micelles with a diameter of 10-15 nm. Targeting integrins on the surface of GBM cells does not appear to improve uptake of the micelles compared to untargeted micelles. TRaM appears to reduce cell viability (~20%) within 24 hours. However, cells require retreatment daily to maintain cell cytotoxicity. Micelles do have an effect on cell viability and may lead to a more concentrated and localized treated of GBM cells.