Surface-engineered dual drug-loaded tumor-targeted liposomal nanoparticles toovercome the therapeutic resistance in glioblastoma multiforme

1.논문 제목: Surface-engineered dual drug-loaded tumor-targeted liposomal nanoparticles to

overcome the therapeutic resistance in glioblastoma multiforme

2.저자명: Ramcharan Singh Angom, Hari Krishnareddy Rachamala, Naga Malleswara Rao Nakka, Vijay Sagar Madamsetty, Paola Suarez-Meade, Beatriz I. Fernandez-Gil, Tanmay Kulkarni, Raegan M. Weil, Shamit Dutta, Enfeng Wang, Santanu Bhattacharya, Krishnendu Pal, Alfredo Quinones-Hinojosa & Debabrata Mukhopadhyay

3.Publish: Communications Medicine, 18 March 2026

4.DOI: https://doi.org/10.1038/s43856-025-01279-7

5.Abstract:

Background) Glioblastoma (GBM) is the most common high-grade primary malignant brain tumor, characterized by a notably poor prognosis. Current treatments for GBM have shown limited effectiveness in improving patient survival, highlighting the urgent need for effective therapeutic strategies. Combination therapy offers significant potential in overcoming resistance by targeting multiple signaling pathways; however, it often comes with increased toxicity compared to monotherapy. Co-encapsulating multiple therapeutic agents into a tumor-targeted drug delivery platform holds promise for overcoming these limitations and improving treatment outcomes.

Methods) We developed a tumor-targeted liposomal nanoformulation (TTL) using phospholipids, cholesterol, DSPE-(PEG)2000-OMe, and a proprietary tumor-targeting peptide (TTP). The TTL was loaded with everolimus (TTL-E), vinorelbine (TTL-V), rapamycin (TTL-R), a combination (TTL-EV), or (TTL-RV). These formulations were tested in vivo on orthotopic GBM mice, combined with temozolomide and radiation. RNA sequencing was performed to identify molecular and transcriptome changes post-treatment.

Results) TTL demonstrated tumor-specific uptake, effectively delivering drugs to GBM tumors. TTL-EV and TTL-RV outperformed single-drug formulations. Radiation combined with TTL-EV/RV improved tumor growth inhibition and survival, while temozolomide provided minimal benefit. Transcriptome analysis revealed differentially expressed genes (DEGs) linked to DNA damage repair, cell cycle, metabolism, and extracellular matrix pathways.

Conclusions) TTL crossed the blood-brain barrier, targeting tumors effectively. Radiation plus TTL-EV/RV enhanced tumor suppression and survival in GBM models. Gene expression analysis identified DEGs related to DNA damage and cell death. Mechanistic studies suggest TTL-EV plus radiation inhibits mTOR/MAPK pathways and sensitizes tumors to radiation. These findings offer a potential approach for improving GBM treatment.

6.한글 초록: Glioblasoma (GBM)는 치료 저항성과 혈뇌장벽 (BBB) 문제로 기존 치료 효과가 제한적인임. 본 연구에서는 종양 표적 펩타이드가 결합된 리포좀 (TTL)에 everolimus, vinorelbine 등의 약물을 탑재하여 전달함. TTL은 BBB를 통과해 종양에 선택적으로 축적되었으며, 특히 이중 약물 조합 (TTL-EV, TTL-RV)이 단일 약물보다 우수한 항종양 효과를 보였음. 방사선과 병용 시 종양 억제 및 생존율이 증가하였고, DNA damage 및 세포사멸 관련 경로 변화가 확인됨.

7.한글 요약

7-1. Introduction: GBM은 가장 공격적인 뇌종양으로 표준 치료 (수술+방사선+TMZ)에도 불구하고예후가 매우 나쁨. BBB로 인해 약물 전달이 제한되며, 치료 저항성이 주요 문제임. 이를 해결하기 위해 본 연구는 종양 표적 리포좀 기반 이중 약물 전달 시스템 (TTL)을 개발하여 병용치료 효과를 평가함.

7-2. Method:

7-2-1) Liposomal nanoparticle 제작: Phospholipid, cholesterol, DSPE-PEG2000, TTP로 구성되며, 탑재된 약물은 TTL-E (everolumus), TTL-V (vinorelbine), TTL-EV (dual drug), TTL-R, TTL-RV 이며, ethanol injection method를 사용함.

7-2-2) 세포 실험 (in vitro): GBM1A, GBM22, QNS120, QNS108 (patient-derived) 세포주를 사용함.

세포 생존율 (IC50), uptake (confocal microscopy, flow cytometry), migration (wound healing assay), colony formation assay를 통해 분석함.

7-2-3) 동물 실험 (in vivo): Orthotopic GBM xenograft mouse 모델을 사용함. TTL formulations (IV

injection, 주 2회), 방사선 치료 (5일 연속 조사), TMZ 병용 비교하여 처리함. Tumor growth, survival,

biodistributions (IVIS, Raman imaging)을 통해 평가함.

7-2-4) 분자 분석: RNA sequencing을 통해 DEG 분석, Western blot을 통해 AKT, ERK, mTOR pathway, DNA damage markers (ATM, ATR, CHK1/2), MGMT, ABCG2 를 확인함.

7-3. Result:

7-3-1) 나노입자 특성: 70-90nm 크기로, +15~21Mv zeta potential, 균일한 구형 구조 (TEM 확인), 30일 이상 안정성 유지하는 것을 확인함.

7-3-2) 종양 표적성 및 BBB 통과: TTL은 control liposome 대비 높은 세포 uptake를 보였고, in vivo 에서 종양 부위 선택적 축적을 확인함. BBB 통과 및 tumor-specific localization 확인함.

7-3-3) 항암 효과 (In vitro): TTL-EV (dual drug)가 단일 약물보다 높은 cytotoxicity를 보였고, TMZ resistant cell line에서도 효과 유지됨. 방사선 병용 시 cytotoxicity 크게 증가함. IC50 감소하여 강한 synergistic effect 확인함.

7-3-4) 종양 성장 억제 및 생존율 (In vivo): TTL-EV, TTL-RV은 단일 약물 효과보다 크게 작용했고 방사선 병용 시 tumor growth 억제 증가하고 survival 증가함. TMZ 병용 효과는 제한적임.

7-3-5) 세포 기능 억제: colony formation 감소하고, migration 및 motility는 억제함. 방사선 병용시

효과는 극대화됨.

7-3-6) 분자 기전: TTL-EV + radiation 처리 시 AKT, ERK (MAPK pathway 억제) 감소, mTOR signaling 감소, MGMT, ABCG2 (약물 저항성 관련) 감소, ATM, ATR, CHK1/2 (DNA damage response 억제) 감소. 결과적으로 DNA damage 증가, 세포 사멸 촉진, 방사선 감수성 증가함.

7-3-7) Transcriptome 분석: DEGs 확인 결과, DNA damage repair, cell cycle, metabolism, extracellular matrix, 치료 반응 관련 pathway 변화 확인.

8.대표 Figure: Fig2. In vitro and In vivo Tumor Uptake Assay.