Joshua Yi Lisa Zhang (2) Facilitation of an orthogonal IL-2 system for CAR Tcell therapy through the novel knockout of the human IL-2 gene

Overview

Milone Lab, University of Pennsylvania Chimeric antigen receptor (CAR) Tcell therapy is a novel immunotherapy that engineers a patient’s own Tcells with an artificial receptor to recognize and attack cancer. While this approach has been successful in treating hematologic malignancies, it has struggled in solid tumors. Adjuvant interleukin-2 (IL-2) can increase Tcell expansion, survival, and function, potentially improving CAR Tcell therapy. However, when administered at high levels, IL-2 therapy induces severe toxicity due to the overactivation of Tcells throughout the body. Furthermore, IL-2 stimulates regulatory Tcells (Tregs) which are immunosuppressive. To overcome these issues, an orthogonal cytokine system was created using a mutated cytokinecytokine receptor pair that functions like the endogenous IL-2 pair but makes use of a unique binding site. This approach ensures that orthogonal IL-2 will specifically interact with CAR Tcells engineered with the orthogonal receptor. Unfortunately, upon activation CAR Tcells still produce endogenous IL-2, which promotes immunosuppressive Tregs. Therefore, this study tested six CRISPR-Cas9 gene editing molecules designed to disrupt the endogenous IL-2 gene, preventing endogenous IL-2 secretion. Here, we show the feasibility of a CRISPR-Cas9 knockout of IL-2 production in human Tcells. Tcells transfected with one of six guide RNAs targeting the IL-2 gene were able to achieve a 97.3% reduction of IL-2 secretion compared to control cells. Data from this study supports the use of this orthogonal IL-2 system, which will hopefully allow for CAR Tcell therapy to serve as a successful cancer treatment with remarkable persistence. Testing The Effect of Different Salt/ Salt Alternatives on Conductivity Using Soil Columns Arsonlove A. Abney duPont Manual High School Louisville, KY Teacher Keri Polevchak The first year this project was done while taking water quality samples - from Broad Run (forested watershed) and Chenoweth Run (urban watershed). The data sho wed that Chenoweth Run had very high conductivity, it ranged from 150 microsiemens per centimeter (uS/cm) higher to 800 us/cm higher than Broad Run. Because of this, it was decided to go upstream of Chenoweth Run to see what was making the conductivity in this urban watershed so much higher than its forested counterpart. When testing twelve different sites upstream of Chenoweth Run it was found out conductivity was higher in urban areas, with impervious surfaces, especially during the winter season. However further data analysis gathered that these urban sites have high conductivity year-round, due to high amounts of salt in groundwater released year-round. Originally data was obtained from four different water quality tests, statistics providing rainfall, an online program providing the percentage of impervious surfaces in the watershed. This year data was obtained using soil columns, a conductivity meter, and a lab analysis of samples. To determine which salt alternative prevented the most damage to watersheds, concerning conductivity the different salt alternatives were tested. It was hypothesized that if the road salt alternative used contains less sodium chloride than road salt, conductivity will be lower. After organizing the data obtained and looking at the trends present it showed that the salt alternatives with low levels of NaCl and high levels of CaMg were better for watersheds.

Awards (1)

• 1st Place Medicine & Health/Behavioral Sciences

Competition history

• JSHS 2020 Category not listed

Resources

• National JSHS abstract book ↗

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