Single-Cell Heterogeneity of EGFR Pathway Is Linked to Unique Signatures of Drug Response and Malignancy in Patient Derived Glioblastoma Stem Cells

Michael D Masterman-Smith; Nicholas A Graham; Eduard H Panosyan; Jack Mottahedeh; Eric R Samuels; Araceli Nunez; Tiffany Phillips; Meeryo Choe; Timothy F Cloughesy; Jorge A Lazareff; Linda M Liau; William H Yong; Thomas G Graeber; Harley I Kornblum; Ming-Fei Lang; Yanzhao Li; Jing Sun

doi:10.4143/crt.2024.859

Single-Cell Heterogeneity of EGFR Pathway Is Linked to Unique Signatures of Drug Response and Malignancy in Patient Derived Glioblastoma Stem Cells

Cancer Res Treat. 2025 Jan 7. doi: 10.4143/crt.2024.859. Online ahead of print.

Authors

Michael D Masterman-Smith^{1

2

3}, Nicholas A Graham^{4

5}, Eduard H Panosyan^{6

7}, Jack Mottahedeh⁸, Eric R Samuels⁹, Araceli Nunez¹⁰, Tiffany Phillips¹¹, Meeryo Choe^{6

12}, Timothy F Cloughesy^{6

12}, Jorge A Lazareff^{7

13}, Linda M Liau¹³, William H Yong¹⁴, Thomas G Graeber^{15

16}, Harley I Kornblum^{7

12

16}, Ming-Fei Lang¹⁷, Yanzhao Li¹⁸, Jing Sun^{1

19}

Affiliations

¹ The First Affiliated Hospital of Jinzhou Medical University, Jinzhou, China.
² Rapid Acceleration of Diagnostics (RADx) COVID-19 Response Program, U.S. National Institutes of Health/National Institutes of Biomedical Imaging and Bioengineering, Bethesda, MD, USA.
³ Creative Sciences, Inc., Los Angels, CA, USA.
⁴ Mork Family Department of Chemical Engineering and Materials Science, University of Southern California, Los Angeles, CA, USA.
⁵ Norris Comprehensive Cancer Center, University of Southern California, Los Angeles, CA, USA.
⁶ Department of Medicine, David Geffen School of Medicine, University of California at Los Angeles, CA, USA.
⁷ Department of Pediatrics, David Geffen School of Medicine, University of California at Los Angeles, CA, USA.
⁸ Department of Psychiatry, David Geffen School of Medicine, University of California at Los Angeles, CA, USA.
⁹ Abbvie Inc., Irvine, CA, USA.
¹⁰ School of Nursing, University of California at Los Angeles, CA, USA.
¹¹ Samuel Oschin Comprehensive Cancer Center, Cedars Sinai Medical Center, Los Angeles, CA, USA.
¹² Department of Neurology, David Geffen School of Medicine, University of California at Los Angeles, CA, USA.
¹³ Department of Neurosurgery, David Geffen School of Medicine, University of California at Los Angeles, CA, USA.
¹⁴ Department of Pathology and Laboratory Medicine, David Geffen School of Medicine, University of California at Los Angeles, CA, USA.
¹⁵ Department of Molecular and Medical Pharmacology, David Geffen School of Medicine, University of California at Los Angeles, CA, USA.
¹⁶ California NanoSystems Institute, University of California at Los Angeles, CA, USA.
¹⁷ College of Medicine, Dalian Key Laboratory of Oligosaccharide Recombination and Recombinant Protein Modification, Dalian University, Dalian, China.
¹⁸ Department of Neurosurgery, Affiliated Zhongshan Hospital of Dalian University, Dalian, China.
¹⁹ College of Environmental and Chemical Engineering, Dalian University, Dalian, China.

PMID: 39788641
DOI: 10.4143/crt.2024.859

Abstract

Purpose: In glioblastoma, the therapeutically intractable and resistant phenotypes can be derived from glioma stem cells, which often have different underlying mechanisms from non-stem glioma cells. Aberrant signaling across the EGFR-PTEN-AKT-mTOR pathways have been shown as common drivers of glioblastoma. Revealing the inter and intra-cellular heterogeneity within glioma stem cell populations in relations to signaling patterns through these pathways may be key to precision diagnostic and therapeutic targeting of these cells.

Materials and methods: Single cell parallel proteomic heterogeneity profiling of the EGFR-PTEN-AKT-mTOR pathways was conducted in a panel of fifteen glioma stem cell models derived from patient glioblastoma biopsies.

Results: The analysis included 59,464 data points from 14,866 cells and identified forty-nine molecularly distinct signaling phenotypes. High content bioinformatics resolved two unique patient clusters diverging on EGFR expression and AKT/TORC1 activation. Phenotypic validation indicated drug responsive phenotypes to EGFR blocking in the high EGFR expressing cluster with lower tumor initiating potential in comparison to the AKT/TORC1 activated cluster. High EGFR expression trended with improved patient prognosis while AKT/TORC1 activated samples trended with poorer patient outcomes. Genetic heterogeneity was observed in both clusters with proneural, classical and mesenchymal subtypes observed.

Conclusion: Quantitative single cell heterogeneity profiling reveals divergent EGFR-PTEN-AKT-mTOR pathways of patient derived glioma stem cells, which would inform future research and personalized therapeutic strategies.

Keywords: EGFR; Microfluidics; Neoplastic stem cells; PI3K pathway; Single-cell analysis; glioblastoma; Gliomaspheres.