Phenotypic screens identify biologic regulators of nanoparticle uptake in diffuse midline glioma
- 1Ben Towne Center, Seattle Childrens Research Institute; Department of Pediatrics, University of Washington; Department of Bioengineering, University of Washingt
Abstract
Nanoparticle drug delivery systems hold considerable promise for locoregional administration to central nervous system tumors, yet the biological determinants of nanoparticle-cancer cell interactions remain poorly understood. Using patient-derived histone-mutant diffuse midline glioma (DMG) models, we performed a pooled CRISPR-Cas9 perturbation screen to systematically identify regulators of liposomal nanoparticle delivery. The screen identified candidate genes spanning endocytosis, vesicle transport, and metabolic signaling, revealing that nanoparticle delivery is governed by a broader landscape than previously appreciated. Among these, CTNNB1, or {beta}-catenin, emerged as a common negative regulator across two independent DMG models and two distinct nanoparticle surface chemistries. Transcriptomic profiling of CTNNB1-depleted DMG cells revealed upregulation of membrane remodeling and extracellular matrix gene programs, accompanied by reduced cell stiffness measured by a microfluidic acoustic scattering assay. This resulted in a shift in endocytic activity characterized by decreased bulk-phase macropinocytosis and increased receptor-mediated endocytosis. We further identified MAPK and mTOR pathway members as nanoparticle trafficking modulators, and demonstrated concordance between genetic and pharmacologic perturbations in modulating the liposomal nanoparticle interactions in pediatric DMG cells. These findings establish a biology-first screening approach for identifying previously unappreciated regulators with potential relevance to nanoparticle-based therapeutic strategies in pediatric brain tumors.
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BibTeX
@misc{cheng2026phenotypic,
title = {{Phenotypic screens identify biologic regulators of nanoparticle uptake in diffuse midline glioma}},
author = {E. L. Cheng and S. Pal and J. W. Tsai and C. B. Yampanis and S. Rangaswamy and J. Kenny-Serrano and M. Coppola and M. Ozdemir and K. Y. Cai and E. VanGosen and E. Bateman and K. R. Bennett and M. M. Billingsley and C. A. Whittaker and Y. Zhang and M. Dada and S. Bhatia and C. Fu and Y. Bae and N. Katiyar and J. G. Doench and K. L. Ligon and S. R. Manalis and P. T. Hammond and P. Bandopadhayay and J. P. Straehla},
howpublished = {bioRxiv},
year = {2026},
month = sep,
doi = {10.64898/2026.09.08.749175},
url = {https://www.biorxiv.org/content/10.64898/2026.09.08.749175v1},
note = {CC BY},
}
RIS
TY - UNPB
TI - Phenotypic screens identify biologic regulators of nanoparticle uptake in diffuse midline glioma
AU - E. L. Cheng
AU - S. Pal
AU - J. W. Tsai
AU - C. B. Yampanis
AU - S. Rangaswamy
AU - J. Kenny-Serrano
AU - M. Coppola
AU - M. Ozdemir
AU - K. Y. Cai
AU - E. VanGosen
AU - E. Bateman
AU - K. R. Bennett
AU - M. M. Billingsley
AU - C. A. Whittaker
AU - Y. Zhang
AU - M. Dada
AU - S. Bhatia
AU - C. Fu
AU - Y. Bae
AU - N. Katiyar
AU - J. G. Doench
AU - K. L. Ligon
AU - S. R. Manalis
AU - P. T. Hammond
AU - P. Bandopadhayay
AU - J. P. Straehla
PY - 2026
DA - 2026/09/08
PB - bioRxiv
DO - 10.64898/2026.09.08.749175
UR - https://www.biorxiv.org/content/10.64898/2026.09.08.749175v1
AB - Nanoparticle drug delivery systems hold considerable promise for locoregional administration to central nervous system tumors, yet the biological determinants of nanoparticle-cancer cell interactions remain poorly understood. Using patient-derived histone-mutant diffuse midline glioma (DMG) models, we performed a pooled CRISPR-Cas9 perturbation screen to systematically identify regulators of liposomal nanoparticle delivery. The screen identified candidate genes spanning endocytosis, vesicle transport, and metabolic signaling, revealing that nanoparticle delivery is governed by a broader landscape than previously appreciated. Among these, CTNNB1, or {beta}-catenin, emerged as a common negative regulator across two independent DMG models and two distinct nanoparticle surface chemistries. Transcriptomic profiling of CTNNB1-depleted DMG cells revealed upregulation of membrane remodeling and extracellular matrix gene programs, accompanied by reduced cell stiffness measured by a microfluidic acoustic scattering assay. This resulted in a shift in endocytic activity characterized by decreased bulk-phase macropinocytosis and increased receptor-mediated endocytosis. We further identified MAPK and mTOR pathway members as nanoparticle trafficking modulators, and demonstrated concordance between genetic and pharmacologic perturbations in modulating the liposomal nanoparticle interactions in pediatric DMG cells. These findings establish a biology-first screening approach for identifying previously unappreciated regulators with potential relevance to nanoparticle-based therapeutic strategies in pediatric brain tumors.
ER -