The Application of a Characterized Pre-Clinical Glioblastoma Oncosphere Model to In vitro and In vivo Therapeutic Testing

Glioblastoma;Whole exome sequencing;High throughput drug screening;Drug combinations;Cell Biology

Wilson, Kelli McDowellCasero, Robert A. (Bob) ;

Johns Hopkins University

关键词: Glioblastoma;    Whole exome sequencing;    High throughput drug screening;    Drug combinations;    Cell Biology;   

Others  :  https://jscholarship.library.jhu.edu/bitstream/handle/1774.2/40653/Kelli%20Wilson%20-%20Complete%20Thesis%20%281%29.docx?sequence=2&isAllowed=y

瑞士|英语

来源: JOHNS HOPKINS DSpace Repository

【 摘 要 】

Glioblastoma multiforme (GBM) is a lethal brain cancer with a median survival time (MST) of approximately 15 months following treatment.A serious challenge facing the development of new drugs for the treatment of GBM is that preclinical models fail to replicate the human GBM phenotype.Here we report the Johns Hopkins Oncosphere Panel (JHOP), a panel of GBM oncosphere cell lines. These cell lines were validated by their ability to form tumors intracranially with histological features of human GBM and GBM variant tumors.We then completed whole exome sequencing on JHOP and found that they contain genetic alterations in GBM driver genes such as PTEN, TP53 and CDKN2A.Two JHOP cell lines were utilized in a high throughput drug screen of 466 compounds that were selected to represent late stage clinical development and a wide range of mechanisms.Drugs that were inhibitory in both cell lines were EGFR inhibitors, NF-kB inhibitors and apoptosis activators.We also examined drugs that were inhibitory in a single cell line.Effective drugs in the PTEN null and NF1 wild type cell line showed a limited number of drug targets with EGFR inhibitors being the largest group of cytotoxic compounds.However, in the PTEN mutant, NF1 null cell line, VEGFR/PDGFR inhibitors and dual PIK3/mTOR inhibitors were the most common effective compounds. Using active compounds from the single agent screen, we subjected two oncosphere cell lines to a drug combination matrix high throughput screen.Synergistic combinations were tested in vivo and a PIK3 inhibitor paired with three different drugs extended survival in the JHH-520 GBM model. This data shows that the JHOP is amenable to a high throughput drug screening format and that future preclinical studies on active compounds found in these results may deliver promising therapeutic leads.