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C. Krona, Soumi Kundu, Emil Rosén, F. Kruse, Madeleine Skeppås, H. Babačić, Ida Larsson, Ludmila Elfineh, M. Lü, Maria Escriva Conde, Ramy Elgendy, Zankruti Dave, Milena Doroszko, Katrin Rut-Halldorsdottir, Xiaofang Cao, R. Ramachandra, K. Olausson, Mats Nilsson, Joachim Weischenfeldt, J. Wikström, M. Pernemalm, A. Sundström, Irem Uppman, H. Mangukiya, Sven Nelander
0 1. 7. 2026.

A Multi-Omic Phenobank Reveals Axes of Glioblastoma Growth, Invasion, and Therapeutic Vulnerability

Background Glioblastoma (GBM) invasion is clinically decisive but difficult to model systematically. Existing patient-derived xenograft (PDX) resources rarely couple reproducible in vivo invasion phenotypes with matched multi-omic profiles at scale, limiting mechanistic insight and phenotype-informed therapeutic hypotheses. Methods We established the HGCC Phenobank, comprising 65 patient-derived GBM stem-like cultures with matched multi-omic profiling and orthotopic engraftment in 449 mice. Blinded histopathology quantified ten invasion traits per case. These phenotypes were integrated with RNA sequencing, DNA methylation, and mass-spectrometry-based proteomics. Multi-Omic Factor Analysis (MOFA) identified latent molecular programs. Phenotype-specific RNA signatures were matched to LINCS drug-perturbation profiles and validated in 3D gliomasphere and ex vivo brain-slice assays. Results Two dominant, reproducible invasion modes emerged across models: diffuse parenchymal infiltration and perivascular/condensed growth. Proneural cultures formed more aggressive tumors in immunodeficient mice, and mouse survival showed a modest correlation with patient survival in matched cases (Pearson 0.1832, 0.045). MOFA identified 15 latent factors; Factor 1, enriched for ASCL1/OLIG1/OLIG2 programs and associated with TP53/DCHS2/WNK2 alterations, was linked to increased tumor formation, diffuse invasion, and shorter mouse survival, and stratified GBM patients in TCGA and in our matched patient cohort. Drug-signature matching separated mechanisms targeting diffuse versus perivascular invasion. Experimental validation confirmed phenotype-selective sensitivities, and inhibitors PIK-75 and buparlisib suppressed invasion dynamics across representative models in 3D and brain-slice assays. Conclusions The HGCC Phenobank provides the first openly available PDX resource that systematically links GBM invasion phenotypes to multi-omic programs and therapeutic predictions. This framework enables reproducible model selection, mechanistic dissection of invasion modes, and phenotype-guided therapeutic discovery. Key Points Diffuse and perivascular invasion define orthogonal GBM axes ASCL1/OLIG factor links initiation, diffuse growth, and survival Phenotype-matched drugs validated; PIK-75 and buparlisib curb invasion dynamics Importance of the Study Glioblastoma invasion varies substantially between patients, yet existing patient-derived xenograft resources rarely combine reproducible in vivo phenotyping with matched multi-omic profiling at scale. The HGCC Phenobank addresses this gap with standardized, blinded scoring of ten invasion traits across 449 orthotopic xenografts from 65 molecularly characterized GBM stem-like cultures, integrated with transcriptomic, methylomic, and proteomic data. We identify two dominant, reproducible invasion modes and a cross-modal neurodevelopmental program, the ASCL1/OLIG1/2-associated Factor 1, that links tumor initiation, diffuse growth, and survival in mice, and stratifies GBM patients in TCGA and in our matched patient cohort. In a spatially resolved xenograft section, Factor 1 signal localizes to the invasive tumor periphery. By matching phenotype-specific RNA signatures to drug-induced transcriptional responses, we show that invasion phenotypes nominate selective vulnerabilities, exemplified by PIK-75. This openly shared resource enables reproducible model selection, mechanistic dissection of invasion programs, and phenotype-guided therapeutic discovery.


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