{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,31]],"date-time":"2026-08-31T18:16:30Z","timestamp":1788200190879,"version":"build-2803163510"},"reference-count":49,"publisher":"MDPI AG","issue":"4","license":[{"start":{"date-parts":[[2023,2,8]],"date-time":"2023-02-08T00:00:00Z","timestamp":1675814400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"California Pacific Medical Center Foundation"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cells"],"abstract":"<jats:p>Glioblastoma\u2019s (GBM) aggressive growth is driven by redundant activation of a myriad of signaling pathways and genomic alterations in tyrosine kinase receptors, such as epidermal growth factor receptor (EGFR), which is altered in over 50% of cases. Single agents targeting EGFR have not proven effective against GBM. In this study, we aimed to identify an effective anti-tumor regimen using pharmacogenomic testing of patient-derived GBM samples, in culture and in vivo. High-throughput pharmacological screens of ten EGFR-driven GBM samples identified the combination of erlotinib (EGFRi) and MLN0128 (a mammalian target of rapamycin inhibitor, or MTORi) as the most effective at inhibiting tumor cell viability. The anti-tumor activity of erlonitib+MLN0128 was synergistic and produced inhibition of the p-EGFR, mitogen-activated protein kinase (MAPK), and Phosphoinositide 3-kinase (PI3K) pathways in culture. Using an orthotopic murine model of GBM, we show that erlotinib+MLN0128 inhibited tumor growth in vivo and significantly prolonged the survival of tumor-bearing mice. Expression profiling of tumor tissues from treated mice revealed a unique gene signature induced by erlotinib+MLN0128, consisting of downregulation of immunosuppressive chemokines in the tumor microenvironment, including C-C motif chemokine ligand 2 (CCL2) and periostin. Lower periostin levels resulted in the inhibition of Iba1+ (tumor-promoting) macrophage infiltration of GBM xenografts. Taken together, our results demonstrate that pharmacological co-targeting of EGFR and MTOR using clinically available drugs represents an effective treatment paradigm for EGFR-driven GBMs, acting both by inhibiting tumor cell growth and modulating the immune tumor microenvironment.<\/jats:p>","DOI":"10.3390\/cells12040547","type":"journal-article","created":{"date-parts":[[2023,2,8]],"date-time":"2023-02-08T04:21:59Z","timestamp":1675830119000},"page":"547","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":21,"title":["Dual Targeting of EGFR and MTOR Pathways Inhibits Glioblastoma Growth by Modulating the Tumor Microenvironment"],"prefix":"10.3390","volume":"12","author":[{"given":"Maxim","family":"Sidorov","sequence":"first","affiliation":[{"name":"California Pacific Medical Center Research Institute, 475 Brannan St, Suite 130, San Francisco, CA 94107, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Pratiksha","family":"Dighe","sequence":"additional","affiliation":[{"name":"California Pacific Medical Center Research Institute, 475 Brannan St, Suite 130, San Francisco, CA 94107, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rinette W. 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(2017). Glioblastoma, Codon Publications.","DOI":"10.15586\/codon.glioblastoma.2017"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"11622","DOI":"10.1038\/s41598-020-68011-4","article-title":"Longer-term (>\/= 2 years) survival in patients with glioblastoma in population-based studies pre- and post-2005: A systematic review and meta-analysis","volume":"10","author":"Poon","year":"2020","journal-title":"Sci. Rep."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"3002","DOI":"10.1158\/0008-5472.CAN-17-2433","article-title":"Activation of the Receptor Tyrosine Kinase AXL Regulates the Immune Microenvironment in Glioblastoma","volume":"78","author":"Sadahiro","year":"2018","journal-title":"Cancer Res."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"1807","DOI":"10.1126\/science.1164382","article-title":"An integrated genomic analysis of human glioblastoma multiforme","volume":"321","author":"Parsons","year":"2008","journal-title":"Science"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"882","DOI":"10.1093\/neuonc\/noq052","article-title":"mTOR signaling in glioblastoma: Lessons learned from bench to bedside","volume":"12","author":"Akhavan","year":"2010","journal-title":"Neuro. Oncol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"98","DOI":"10.1016\/j.ccr.2009.12.020","article-title":"Integrated Genomic Analysis Identifies Clinically Relevant Subtypes of Glioblastoma Characterized by Abnormalities in PDGFRA, IDH1, EGFR, and NF1","volume":"17","author":"Verhaak","year":"2010","journal-title":"Cancer Cell"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"462","DOI":"10.1016\/j.cell.2013.09.034","article-title":"The somatic genomic landscape of glioblastoma","volume":"155","author":"Brennan","year":"2013","journal-title":"Cell"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"29","DOI":"10.1016\/j.drup.2019.04.002","article-title":"The TICking clock of EGFR therapy resistance in glioblastoma: Target Independence or target Compensation","volume":"43","author":"Saleem","year":"2019","journal-title":"Drug Resist. Updates"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1543","DOI":"10.20892\/j.issn.2095-3941.2022.0540","article-title":"Targeting the EGF receptor family in non-small cell lung cancer-increased complexity and future perspectives","volume":"19","author":"Boch","year":"2022","journal-title":"Cancer Biol. Med."},{"key":"ref_10","first-page":"CD007047","article-title":"Epidermal growth factor receptor (EGFR) inhibitors for metastatic colorectal cancer","volume":"6","author":"Chan","year":"2017","journal-title":"Cochrane. Database Syst. Rev."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"518","DOI":"10.1038\/s41586-021-04393-3","article-title":"Glioblastoma mutations alter EGFR dimer structure to prevent ligand bias","volume":"602","author":"Hu","year":"2022","journal-title":"Nature"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"1061","DOI":"10.1093\/neuonc\/noq072","article-title":"Safety and efficacy of erlotinib in first-relapse glioblastoma: A phase II open-label study","volume":"12","author":"Yung","year":"2010","journal-title":"Neuro Oncol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"413","DOI":"10.1007\/s11060-013-1316-y","article-title":"Efficacy of erlotinib in patients with relapsed gliobastoma multiforme who expressed EGFRVIII and PTEN determined by immunohistochemistry","volume":"116","author":"Gallego","year":"2014","journal-title":"J. Neurooncol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"984","DOI":"10.1093\/neuonc\/nou029","article-title":"A single-institution phase II trial of radiation, temozolomide, erlotinib, and bevacizumab for initial treatment of glioblastoma","volume":"16","author":"Clarke","year":"2014","journal-title":"Neuro Oncol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"295","DOI":"10.1158\/1078-0432.CCR-17-0963","article-title":"Prospective Feasibility Trial for Genomics-Informed Treatment in Recurrent and Progressive Glioblastoma","volume":"24","author":"Byron","year":"2018","journal-title":"Clin. Cancer Res."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"9230479","DOI":"10.1155\/2018\/9230479","article-title":"Targeting mTOR in Glioblastoma: Rationale and Preclinical\/Clinical Evidence","volume":"2018","author":"Mecca","year":"2018","journal-title":"Dis. Markers"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"110","DOI":"10.1016\/j.canlet.2017.04.019","article-title":"The dual mTOR kinase inhibitor TAK228 inhibits tumorigenicity and enhances radiosensitization in diffuse intrinsic pontine glioma","volume":"400","author":"Miyahara","year":"2017","journal-title":"Cancer Lett."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"1073","DOI":"10.1093\/neuonc\/noaa106","article-title":"Glioblastoma in adults: A Society for Neuro-Oncology (SNO) and European Society of Neuro-Oncology (EANO) consensus review on current management and future directions","volume":"22","author":"Wen","year":"2020","journal-title":"Neuro Oncol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"281","DOI":"10.1007\/s11940-013-0226-9","article-title":"Treatment options in newly diagnosed glioblastoma","volume":"15","author":"Lee","year":"2013","journal-title":"Curr. Treat Options Neurol."},{"key":"ref_20","first-page":"92","article-title":"Characterizing Glioblastoma Heterogeneity via Single-Cell Receptor Quantification","volume":"6","author":"Chen","year":"2018","journal-title":"Front. Genet."},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"Ketchen, S.E., Gamboa-Esteves, F.O., Lawler, S.E., Nowicki, M.O., Rohwedder, A., Knipp, S., Prior, S., Short, S.C., Ladbury, J.E., and Bruning-Richardson, A. (2021). Drug Resistance in Glioma Cells Induced by a Mesenchymal-Amoeboid Migratory Switch. Biomedicines, 10.","DOI":"10.3390\/biomedicines10010009"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"648","DOI":"10.1038\/s41416-019-0696-y","article-title":"Drug responses are conserved across patient-derived xenograft models of melanoma leading to identification of novel drug combination therapies","volume":"122","author":"Ice","year":"2020","journal-title":"Br. J. Cancer"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"3065","DOI":"10.1158\/0008-5472.CAN-14-3307","article-title":"Cytomegalovirus Immediate-Early Proteins Promote Stemness Properties in Glioblastoma","volume":"75","author":"Soroceanu","year":"2015","journal-title":"Cancer Res."},{"key":"ref_24","first-page":"vdac019","article-title":"Cannabidiol inhibits RAD51 and sensitizes glioblastoma to temozolomide in multiple orthotopic tumor models","volume":"4","author":"Soroceanu","year":"2022","journal-title":"Neurooncol. Adv."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"1119","DOI":"10.3389\/fonc.2019.01119","article-title":"Tissues Harvested Using an Automated Surgical Approach Confirm Molecular Heterogeneity of Glioblastoma and Enhance Specimen\u2019s Translational Research Value","volume":"9","author":"Zusman","year":"2019","journal-title":"Front. Oncol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"621","DOI":"10.1124\/pr.58.3.10","article-title":"Theoretical basis, experimental design, and computerized simulation of synergism and antagonism in drug combination studies","volume":"58","author":"Chou","year":"2006","journal-title":"Pharmacol. Rev."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"1011173","DOI":"10.3389\/fonc.2022.1011173","article-title":"BPTF promotes the progression of distinct subtypes of breast cancer and is a therapeutic target","volume":"12","author":"Bezrookove","year":"2022","journal-title":"Front. Oncol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"e1601","DOI":"10.1038\/cddis.2014.566","article-title":"Reactive oxygen species-mediated therapeutic response and resistance in glioblastoma","volume":"6","author":"Singer","year":"2015","journal-title":"Cell Death Amp Dis."},{"key":"ref_29","doi-asserted-by":"crossref","unstructured":"Brennan, C., Momota, H., Hambardzumyan, D., Ozawa, T., Tandon, A., Pedraza, A., and Holland, E. (2009). Glioblastoma subclasses can be defined by activity among signal transduction pathways and associated genomic alterations. PLoS ONE, 4.","DOI":"10.1371\/journal.pone.0007752"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"743","DOI":"10.1093\/neuonc\/nox191","article-title":"EGFR heterogeneity and implications for therapeutic intervention in glioblastoma","volume":"20","author":"Eskilsson","year":"2018","journal-title":"Neuro Oncol."},{"key":"ref_31","first-page":"97","article-title":"Synergistic combination of microtubule targeting anticancer fludelone with cytoprotective panaxytriol derived from panax ginseng against MX-1 cells in vitro: Experimental design and data analysis using the combination index method","volume":"6","author":"Zhang","year":"2016","journal-title":"Am. J. Cancer Res."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"391","DOI":"10.1038\/nature07209","article-title":"Platelet-derived growth factor-alpha receptor activation is required for human cytomegalovirus infection","volume":"455","author":"Soroceanu","year":"2008","journal-title":"Nature"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"105780","DOI":"10.1016\/j.phrs.2021.105780","article-title":"Glioblastoma multiforme (GBM): An overview of current therapies and mechanisms of resistance","volume":"171","author":"Wu","year":"2021","journal-title":"Pharmacol. Res."},{"key":"ref_34","doi-asserted-by":"crossref","unstructured":"Cheng, W.L., Feng, P.H., Lee, K.Y., Chen, K.Y., Sun, W.L., Van Hiep, N., Luo, C.S., and Wu, S.M. (2021). The Role of EREG\/EGFR Pathway in Tumor Progression. Int. J. Mol. Sci., 22.","DOI":"10.3390\/ijms222312828"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"326","DOI":"10.1016\/j.ccell.2017.02.009","article-title":"The Microenvironmental Landscape of Brain Tumors","volume":"31","author":"Quail","year":"2017","journal-title":"Cancer Cell"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"157","DOI":"10.1016\/j.ccr.2006.02.019","article-title":"Molecular subclasses of high-grade glioma predict prognosis, delineate a pattern of disease progression, and resemble stages in neurogenesis","volume":"9","author":"Phillips","year":"2006","journal-title":"Cancer Cell"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"1004","DOI":"10.3389\/fimmu.2018.01004","article-title":"Immune Microenvironment in Glioblastoma Subtypes","volume":"9","author":"Chen","year":"2018","journal-title":"Front. Immunol."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"20","DOI":"10.1038\/nn.4185","article-title":"The role of microglia and macrophages in glioma maintenance and progression","volume":"19","author":"Hambardzumyan","year":"2016","journal-title":"Nat. Neurosci."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"6785","DOI":"10.1158\/0008-5472.CAN-17-3551","article-title":"EGFR Cooperates with EGFRvIII to Recruit Macrophages in Glioblastoma","volume":"78","author":"An","year":"2018","journal-title":"Cancer Res."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"1399","DOI":"10.1016\/j.celrep.2017.10.030","article-title":"Single-Cell RNA-Seq Analysis of Infiltrating Neoplastic Cells at the Migrating Front of Human Glioblastoma","volume":"21","author":"Darmanis","year":"2017","journal-title":"Cell Rep."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"170","DOI":"10.1038\/ncb3090","article-title":"Periostin secreted by glioblastoma stem cells recruits M2 tumour-associated macrophages and promotes malignant growth","volume":"17","author":"Zhou","year":"2015","journal-title":"Nat. Cell Biol."},{"key":"ref_42","doi-asserted-by":"crossref","unstructured":"Lim, M., Puttick, S., Houston, Z.H., Thurecht, K.J., Kalita-de Croft, P., Mahler, S., Rose, S.E., Jeffree, R.L., Mazzieri, R., and Dolcetti, R. (2019). Innovative Therapeutic Strategies for Effective Treatment of Brain Metastases. Int. J. Mol. Sci., 20.","DOI":"10.3390\/ijms20061280"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"1289","DOI":"10.1093\/neuonc\/not093","article-title":"Erlotinib resistance in EGFR-amplified glioblastoma cells is associated with upregulation of EGFRvIII and PI3Kp110delta","volume":"15","author":"Schulte","year":"2013","journal-title":"Neuro Oncol."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"1331","DOI":"10.1093\/neuonc\/now162","article-title":"In search of a target: PD-1 and PD-L1 profiling across glioma types","volume":"18","author":"Berghoff","year":"2016","journal-title":"Neuro Oncol."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"1781","DOI":"10.1016\/j.apsb.2021.12.019","article-title":"Therapeutic strategies of glioblastoma (GBM): The current advances in the molecular targets and bioactive small molecule compounds","volume":"12","author":"Liu","year":"2022","journal-title":"Acta Pharm. Sin. B"},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"750675","DOI":"10.3389\/fgene.2021.750675","article-title":"Immunotherapy Resistance in Glioblastoma","volume":"12","author":"Wang","year":"2021","journal-title":"Front. Genet."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"122","DOI":"10.3389\/fphys.2013.00122","article-title":"YKL-40 acts as an angiogenic factor to promote tumor angiogenesis","volume":"4","author":"Shao","year":"2013","journal-title":"Front. Physiol."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"e147552","DOI":"10.1172\/JCI147552","article-title":"Chitinase-3-like 1 protein complexes modulate macrophage-mediated immune suppression in glioblastoma","volume":"131","author":"Chen","year":"2021","journal-title":"J. Clin. Investig."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"1399","DOI":"10.1038\/s41588-018-0209-6","article-title":"Pharmacogenomic landscape of patient-derived tumor cells informs precision oncology therapy","volume":"50","author":"Lee","year":"2018","journal-title":"Nat. Genet."}],"container-title":["Cells"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2073-4409\/12\/4\/547\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T18:27:52Z","timestamp":1760120872000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2073-4409\/12\/4\/547"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,2,8]]},"references-count":49,"journal-issue":{"issue":"4","published-online":{"date-parts":[[2023,2]]}},"alternative-id":["cells12040547"],"URL":"https:\/\/doi.org\/10.3390\/cells12040547","relation":{},"ISSN":["2073-4409"],"issn-type":[{"value":"2073-4409","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,2,8]]}}}