(T1230-03-18) Targeting Brain Metastasis in Non-Small Cell Lung Cancer with EGFRT790M Mutation Using BBB-Penetrating Codelivery Liposomes

Purpose: Epidermal Growth Factor Receptor (EGFR) TKI therapy has emerged as the primary treatment approach for non-small cell lung cancer (NSCLC) patients harboring EGFR mutations1-2. However, two significant hurdles impede its effectiveness: secondary EGFR mutation-associated TKI resistance and the occurrence of brain metastasis (BMs).3-4  Brain metastasis is a leading cause of mortality among advanced NSCLC patients, and managing BMs with TKI resistance poses a substantial clinical challenge.

Methods: Targeting tumor-associated macrophages (TAM) presents a potential approach for addressing challenges in EGFR TKI therapy for non-small cell lung cancer (NSCLC) patients with EGFR mutations and brain metastasis (BMs). We developed a dual-targeting liposomal system incorporating anti-PD-L1 nanobody and transferrin receptor (TfR)-binding peptide T12 to co-deliver simvastatin and gefitinib for BMs treatment.

Results: Our dual-targeting liposomes effectively traversed the blood-brain barrier (BBB) and reached BMs sites, inducing repolarization of TAM and overcoming EGFR T790M-mediated drug resistance. Treatment mechanisms involved increased reactive oxygen species (ROS) levels and suppression of the EGFR/Akt/Erk signaling pathway.

Conclusion: This dual-targeting liposomal co-delivery system presents a promising therapeutic strategy for addressing the challenges faced by advanced NSCLC patients with EGFR T790M mutations and BMs.

References: 1. Nan X, Xie C, Yu X, Liu J. EGFR TKI as first-line treatment for patients with advanced EGFR mutation-positive non-small-cell lung cancer. Oncotarget. 2017;8:75712–26.
2. Shi Y, Au JS, Thongprasert S, Srinivasan S, Tsai CM, Khoa MT. et al. A prospective, molecular epidemiology study of EGFR mutations in Asian patients with advanced non-small-cell lung cancer of adenocarcinoma histology (PIONEER) J Thorac Oncol. 2014;9:154–62.
3. Westover D, Zugazagoitia J, Cho BC, Lovly CM, Paz-Ares L. Mechanisms of acquired resistance to first- and second-generation EGFR tyrosine kinase inhibitors. Ann Oncol. 2018;29:i10–i19.
4. Ghosh A, Yan H. Hydrogen bond analysis of the EGFR-ErbB3 heterodimer related to non-small cell lung cancer and drug resistance. J Theor Biol. 2019;464:63–71.

Acknowledgements: This work was supported by NFSC (81925035, 81673382 and 81521005), and the Strategic Priority Research Program of CAS (XDA12050307)


The characterizations of the T12/P-Lipo. (A) The particle size and TEM of the T12/P-Lipo. (B) Transwell co-culture model. (C) The targeting receptors (TfR and PD-L1) in the BCEC w/ or w/o co-culture of H1975 cells, as well as in the H1975 cells alone. (D) Uptake efficiency of Lipo, P-Lipo, T12-Lipo, and T12/P-Lipo in the H1975 cells. (E) Uptake of Lipo, P-Lipo T12-Lipo and T12/P-Lipo in the H1975 cells after penetration through the BCEC monolayer at the Transwell. (F) Fluorescence images of the intracellular Lipo, P-Lipo T12-Lipo, and T12/P-Lipo in the tumor cells. (G) Fluorescence images of the intracellular Lipo, P-Lipo, T12-Lipo, and T12/P-Lipo in the H1975 cells after penetration through the BCEC monolayer at the Transwell.


Biodistribution study of T12/P-Lipo. (A) The intratumor accumulation of T12/P-Lipo. (B) In vivo radiant efficiency of ROIs at tumor sites. (C) In vitro radiant efficiency of the organs. (D) Radiant efficiency of the dissected brain tissues. (E) Ex vivo relative radiant intensity in the brains. (F) The expression of PD-L1 and TfR in the BMs and PD-L1 in M2Φ. (G) Immunofluorescent co-localization (the arrows) of the DIR-containing liposomes and PD-L1 or TfR.


The treatment efficacy in the BMs model of H1975 NSCLC. (A) The treatment regimen. (B) The tumor location in brain tissue (H&E result). (C) The survival curves. (D) The median survival. (E) The histological examination of the brain tissues with tumor xenografts after treatment. (F) Statistical analysis of tumor regression after treatment. (G) Downregulation of p-EGFR/p-Akt/p-Erk. (H) Downregulation of caspase 3 and Ki 67. (I) The M2-associated CD206 expression. (J) The enhanced apoptosis (TUNEL staining) and reduced proliferation (Ki 67 staining) (brown color) in tumors. (K) M2-associated marker CD206 and M1-associated marker iNOS staining after treatment.