Abstract 1102: Establishing the cytotoxic benefit of Tumor Treating Fields on radiation sensitive and acquired radiation resistant glioblastoma patient derived xenograft pairs
Authors: Schanel Taylor L. , Jones Amber B. , Beierle Andee M. , Alrefai Hasan , Nassour Lauren C. , Hicks Patricia H. , Anderson Joshua C. , Hjelmeland Anita B. , Willey Christopher D. 
Published: April 2023

Background: Glioblastoma (GBM) is the most common malignant primary brain tumor with very poor prognosis. The current standard of care for GBM consists of maximal safe tumor resection, chemotherapy (temozolomide) and radiotherapy followed by adjuvant temozolomide and Tumor Treating Fields (TTFields). Unfortunately, GBM displays both inherent and acquired resistance to temozolomide and radiation and is incurable. While TTFields have been found to broadly induce endoplasmic reticulum stress, autophagy, cell membrane permeability, DNA replication stress, anti-mitotic and anti-migratory effects, it is unclear how well TTFields impact radiation resistant GBM. To better understand the cytotoxic benefit of TTFields on acquired radiation resistant GBM, we tested patient-derived xenograft (PDX) brain tumor initiating cells (BTICs) that had been selected for radiation resistance and compared them to their radiation sensitive parent.

Methods: PDX were developed with subcutaneous implantation of patient GBM cells into immunodeficient (nu/nu) mice. Acquired radiation resistant PDXs were generated through in vivo selection (6-8 passages) using six fractions of 2 Gray of radiation over two weeks. PDX BTICs were cultured on laminin-coated cover slips under serum free conditions. Cytotoxicity was measured on both the two-dimensional sham control and two-dimensional TTFields treatment conditions over the course of 72 hours and analyzed with crystal violet staining for both imaging and absorbance quantification. PamStation 12 Kinomic data and RNA-seq (Illumina) data was generated for each PDX.

Results: Crystal violet quantification and representative imaging indicated that TTFields inhibit cell growth in both parental and acquired radiation resistant PDX-derived BTICs. There is an observed TTFields sensitivity response from the acquired radiation resistant PDX, indicating that TTFields treatment does have a cytotoxic effect on acquired radiation resistant models (seen over four reproducible experiments). Baseline kinomic and transcriptomic differences between the PDX lines suggest potential TTFields sensitivity signatures.

Conclusions: Ongoing studies are focused on validation of this cytotoxic effect on larger cohort of both acquired and inherent radiation resistant PDX cells. We plan to measure TTFields -treatment induced kinomic alterations to identify downstream signaling pathways associated with TTFields treatment response, as well as the development of acquired TTFields resistant GBM PDX cells.

Citation Format: Taylor L. Schanel, Amber B. Jones, Andee M. Beierle, Hasan Alrefai, Lauren C. Nassour, Patricia H. Hicks, Joshua C. Anderson, Anita B. Hjelmeland, Christopher D. Willey. Establishing the cytotoxic benefit of Tumor Treating Fields on radiation sensitive and acquired radiation resistant glioblastoma patient derived xenograft pairs [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 1102.