TMOD-36. NANOGLIOS: A MINIATURIZED PATIENT AVATAR ENABLING RAPID AND HIGH-THROUGHPUT PHENOTYPIC SCREENING FOR GLIOMAS

Abstract BACKGROUND Establishing pre-clinical patient derived orthtopical xenograft models for IDH-mutant and H3K27M-mutant gliomas has presented significant histological challenges. There is increasing interest in developing faithful patient-derived ex vivo 3D models to replicate these type of gliomas, aiming to accelerate drug screening efforts. METHOD We developed a rapid approach for establishing nanoliter-sized miniature glioma organoids, termed NanoGlios, by leveraging emulsion microfluidics. We tested the robustness of NanoGlios for neurosphere culture, glioma xenografts, and primary patient samples, focusing on H3K27M-mutant diffuse midline gliomas (DMGs) and IDH-mutant gliomas. We compared the morphological features of NanoGlios to their counterparts using high-content imaging, H&E staining, and histological analysis. Additionally, we evaluated the mutation and gene expression profiles of NanoGlios relative to their counterparts. Furthermore, we conducted proof-of-concept drug screening of NanoGlios with various inhibitors and established an AI-enhanced high-throughput phenotypic imaging-based screening platform. RESULTS We successfully established NanoGlios from various sample sources, including neurospheres, xenografts, and primary tissues harboring H3K27M or IDH1/2 mutations. NanoGlios derived from xenografts or primary tissues captured more heterogeneous patterns as compared to the neuropsheres. We were able to generate thousands of NanoGlios and perform drug assays for hundreds of conditions at passage 0 within two weeks from both resected glioma tissues and xenografts. The AI phenotypic screening enabled us to identify differential responses of NanoGlios to ONC-201 treatment and drug responses to mutant IDH inhibitors Vorasidenib and Ivosidenib, which was not able to be detected by conventional cell viability bulk assays.