entrectinib and Glioma

Topics: Benzamides; Child; Glioma; Humans; Indazoles; Rho Guanine Nucleotide Exchange Factors; Spinal Cord

Topics: Benzamides; Carrier Proteins; Glioma; Humans; Indazoles; Infant; Lung Neoplasms; Nuclear Proteins; Protein-Tyrosine Kinases; Proto-Oncogene Proteins

To accurately recapitulate the heterogeneity of human diseases, animal models require to recreate multiple complex genetic alterations. Here, we combine the RCAS-TVA system with the CRISPR-Cas9 genome editing tools for precise modeling of human tumors. We show that somatic deletion in neural stem cells of a variety of known tumor suppressor genes (Trp53, Cdkn2a, and Pten) leads to high-grade glioma formation. Moreover, by simultaneous delivery of pairs of guide RNAs we generate different gene fusions with oncogenic potential, either by chromosomal deletion (Bcan-Ntrk1) or by chromosomal translocation (Myb-Qk). Lastly, using homology-directed-repair, we also produce tumors carrying the homologous mutation to human BRAF V600E, frequently identified in a variety of tumors, including different types of gliomas. In summary, we have developed an extremely versatile mouse model for in vivo somatic genome editing, that will elicit the generation of more accurate cancer models particularly appropriate for pre-clinical testing.

Topics: Animals; Antigens, Neoplasm; Benzamides; Brain Neoplasms; Brevican; CRISPR-Cas Systems; DNA Repair; False Positive Reactions; Gene Editing; Gene Frequency; Gene Transfer Techniques; Glioma; Humans; In Situ Hybridization, Fluorescence; Indazoles; Mice; Mice, SCID; Mice, Transgenic; Mutation; NIH 3T3 Cells; Receptor, trkA; RNA, Guide, Kinetoplastida

The widespread application of high-throughput sequencing methods is resulting in the identification of a rapidly growing number of novel gene fusions caused by tumour-specific chromosomal rearrangements, whose oncogenic potential remains unknown. Here we describe a strategy that builds upon recent advances in genome editing and combines ex vivo and in vivo chromosomal engineering to rapidly and effectively interrogate the oncogenic potential of genomic rearrangements identified in human brain cancers. We show that one such rearrangement, an microdeletion resulting in a fusion between Brevican (BCAN) and Neurotrophic Receptor Tyrosine Kinase 1 (NTRK1), is a potent oncogenic driver of high-grade gliomas and confers sensitivity to the experimental TRK inhibitor entrectinib. This work demonstrates that BCAN-NTRK1 is a bona fide human glioma driver and describes a general strategy to define the oncogenic potential of novel glioma-associated genomic rearrangements and to generate accurate preclinical models of this lethal human cancer.

Topics: Animals; Benzamides; Brevican; CRISPR-Cas Systems; Drug Screening Assays, Antitumor; Feasibility Studies; Female; Gene Editing; Glioma; Humans; Indazoles; Mice, Inbred C57BL; Mice, Nude; Neoplasms, Experimental; Oncogene Fusion; Primary Cell Culture; Receptor, trkA