Efficient Delivery of Lomitapide using Hybrid Membrane-Coated Tetrahedral DNA Nanostructures for Glioblastoma Therapy

Mingming Song et al. Advanced Materials, 2024

DOI: 10.1002/adma.202311760

Figure 2. Preparation and characterization of RFA NPs

Abstract (원문 요약)

Glioblastoma (GBM) is the most aggressive and prevalent primarymalignant tumor of the central nervous system. Traditional chemotherapy haspoor therapeutic effects and significant side effects due to drug resistance, thenatural blood-brain barrier (BBB), and nonspecific distribution, leading to a lackof clinically effective therapeutic drugs. Here, 1430 small molecule compoundsare screened based on a high-throughput drug screening platform anda novel anti-GBM drug, lomitapide (LMP) is obtained. Furthermore, a bionicnanodrug delivery system (RFA NPs) actively targeting GBM is constructed,which mainly consists of tetrahedral DNA nanocages (tFNA NPs) loaded withLMP as the core and a folate-modified erythrocyte–cancer cell–macrophagehybrid membrane (FRUR) as the shell. FRUR camouflage conferredunique features on tFNA NPs, including excellent biocompatibility, improvedpharmacokinetic profile, efficient BBB permeability, and tumor targetingability. The results show that the LMP RFA NPs exhibited superior and specificanti-GBM activities, reduced off-target drug delivery, prolonged lifespan,and has negligible side effects in tumor-bearing mice. This study combineshigh-throughput drug screening with biomimetic nanodrug delivery systemtechnology to provide a theoretical and practical basis for drug developmentand the optimization of clinical treatment strategies for GBM treatment

국문 초록 요약

본 연구는 기존 고지혈증 치료제로 승인된 Lomitapide (LMP)를 기반으로 교모세포종(Glioblastoma, GBM)에 대한 신규 치료제를 개발하고, 이를 효과적으로 표적 전달할 수 있는 바이오미메틱 나노전달 시스템을 구축하였다. 연구팀은 총 1,430종의 소분자 약물 라이브러리를 고속 스크리닝하여 LMP를 항-GBM 후보물질로 선정하였다. LMP는 GBM 세포에서 선택적인 세포독성을 보였으며, apoptosis 뿐만 아니라 pyroptosis (염증성 프로그램성 세포사멸)를 유도하는 특징이 확인되었다. 약물 전달을 위해 테트라헤드랄 DNA 나노구조체(tFNA)를 기반으로 적혈구, 종양세포, 대식세포의 하이브리드 세포막을 코팅하고, 추가로 folic acid을 부착한 RFA 나노입자 (RFA NPs)를 개발하였다. 해당 전달체는 혈뇌장벽(BBB) 통과, 면역회피, 종양 표적화에서 우수한 성능을 보였으며, 체내 독성 또한 최소화되었다. 마우스 뇌종양 모델에서 LMP RFA NPs는 종양 성장 억제 및 생존기간 연장에서 탁월한 치료효과를 입증하였다. RNA-seq 기반 기전 분석을 통해 LMP가 염증성 사이토카인 발현을 유도하여 pyroptosis를 활성화함과 동시에, MMP3 (matrix metalloproteinase-3) 활성을 억제함으로써 종양 증식 및 전이를 저해하는 이중 기전이 확인되었다. 본 연구는 기존 승인약물의 재창출(drug repurposing)과 바이오미메틱 DNA 나노전달 기술을 융합하여 GBM 치료의 한계를 극복할 수 있는 새로운 전략적 플랫폼을 제시하였다.

배경

• GBM은 기존 항암제의 효과가 낮고, 치료 후 재발이 잦으며, 혈뇌장벽(BBB)으로 인해 약물 전달에 한계 존재.

• 기존 GBM 연구는 주로 신규 표적 발굴에 집중되었으나, 본 연구는 기존 승인 약물 재창출(drug repurposing) 접근을 활용.

• 특히, 약물 전달 시스템의 한계 (BBB 통과, 면역 회피, 약물 안정성 부족) 를 극복하기 위한 신규 나노전달 기술 개발이 필요함.

주요 연구 질문

1. 기존 승인 약물 중 GBM 억제 활성을 가진 약물을 발굴할 수 있는가?

2. BBB 통과 및 종양 타겟팅을 동시에 만족하는 나노전달 시스템을 설계할 수 있는가?

3. 신규 약물의 GBM 억제 기전은 무엇인가?

연구 방법

1. 고속 스크리닝 기반 약물 발굴

• 1,430개 소분자 라이브러리 사용

• 2D GBM 세포 (U251)에서 1차 스크리닝 → 70% 이상 세포사멸 유도 약물 선별

• 3D spheroid 기반 U251, U118 모델로 후보 약물 재검증

2. 약물전달 시스템 개발

• 코어: Tetrahedral DNA nanostructure (tFNA) → 구조적 안정성, 생분해성 우수

• 쉘: Hybrid membrane (적혈구+GBM세포+대식세포막)

• 타겟팅: Folate modification (FA) → GBM 세포의 folate receptor 활용

• 최종 전달체: RFA NPs (Hybrid Membrane-Coated DNA Nanostructures)

3. in vitro / in vivo 효능 평가

• 2D/3D spheroid 세포 실험

• Orthotopic GBM mouse 모델 (U251-LUC)

• BBB 통과성, 세포 흡수, 면역 회피성, 생존율 등 평가

4. 작용기전 규명

• RNA-seq 기반 차등발현 유전자 분석

• siRNA knockdown (MTTP)

• Molecular docking (MAPK8, MMP3)

• 효소 저해 활성 검증

주요 결과

[약물 발굴 결과]

• Lomitapide (LMP) → 최종 후보 선정

• 기존에는 고지혈증 치료용 MTTP inhibitor

• GBM 선택적 세포독성 보임

[약물전달 시스템 성능]

지표	결과
Particle size	~35 nm
Encapsulation efficiency	86.5%
Drug loading	35%
BBB 통과성	In vitro / In vivo 모두 확인
Immune escape	macrophage phagocytosis 억제 확인
Hemocompatibility	안정적

[in vitro 효능]

• IC50 감소 (free LMP 5.8 μM → RFA NP 5.0 μM)

• apoptosis 및 pyroptosis 동시 유도

• 3D spheroid 모델에서 종양 성장 완전 억제

[in vivo 효능 (mouse model)]

• Tumor volume 현저히 감소

• 생존기간 유의미한 증가

• 간/신장 기능, 체중 변화, 면역반응 (TNF-α 등) 문제 없음

• 장기 독성 및 염증반응 minimal

[작용기전]

• Pyroptosis 유도: IL-1β, IL-6, GDF15 증가 → 염증성 세포사멸

• MMP3 억제: RNA-seq → hub gene 분석 → docking 및 효소저해 실험으로 확인

• 기존 MTTP 억제는 GBM 세포독성과 무관

결론

• 기존 FDA 승인 약물인 Lomitapide를 GBM pyroptosis 유도제 + MMP3 억제제로 repurposing 성공.

• Hybrid membrane + tFNA 기반 전달 시스템 개발로 BBB 통과, 면역회피, 종양 타겟팅, 안정성 확보.

• 신규 drug delivery platform으로써 임상전이 가능성(clinical translation potential) 보유.

임상적 의의

• 기존 TMZ (Temozolomide) 내성 환자에서 대체 요법으로 활용 가능성

• GBM 이외 solid tumor에도 platform 기술로 응용 가능

• AI 기반 약물 재창출 + DNA origami 기반 DDS의 융합 사례