PO.TB10.17 · 肿瘤生物学

以抑制层粘连蛋白-411 的多功能纳米药物治疗胶质母细胞瘤可激活局部免疫应答、提高小鼠生存并揭示 AI 筛选的免疫分子通路

Glioblastoma treatment with multifunctional nanodrug inhibiting laminin-411 activates local immune response, increases survival of mice and reveals AI-selected immune molecular pathways

编号 3508 展板 1 时间 4/20 02:00–05:00 区域 Section 32 主讲 Alexander Ljubimov, DSc;PhD

分会场 Therapeutic Modulation of the Tumor Microenvironment: New Targets and Approaches 1

该海报暂无可下载的资料 AACR 官方页面

作者与单位 Authors & Affiliations

Alexander V. Ljubimov¹, Rameshwar Patil², Vladimir A. Ljubimov³, Hui Ding³, Yizhou Wang⁴, Sarah Song⁴, Andrei A. Kramerov¹, Oksana Chepurna¹, Vanessa Borges¹, Jessica Dos Santos¹, Eggehard Holler⁵, Julia Y. Ljubimova⁵, Keith L. Black³

¹Biomedical Sciences, Cedars-Sinai Health Sciences University, Los Angeles, CA,²Neurosurgery, Loma Linda University, Loma Linda, CA,³Neurosurgery, Cedars-Sinai Health Sciences University, Los Angeles, CA,⁴Computational Biomedicine, Cedars-Sinai Health Sciences University, Los Angeles, CA,⁵Terasaki Institute for Biomedical Innovation, Los Angeles, CA

摘要 Abstract

中文摘要

引言：癌症微环境影响肿瘤生长、侵袭及免疫监视逃逸。胶质母细胞瘤（GBM）的细胞外基质（ECM），尤其是层粘连蛋白，在肿瘤进展中发挥重要作用。我们此前在来自 130 例患者的 GBM 样本中观察到，ECM 层粘连蛋白-411（alpha4beta1gamma1）表达增加与肿瘤更快复发及患者生存降低之间存在相关性。层粘连蛋白-411 是一种基底膜成分，也可通过 Notch 信号调节免疫应答。我们开发了一种阻断层粘连蛋白-411 的新型纳米偶联物（NC），可在脑内递送后激活 GBM 局部免疫系统。方法：NC 以无毒的聚（beta-L-苹果酸，P）为骨架，共价连接了针对层粘连蛋白-411 alpha4 和 beta1 链的反义寡核苷酸（AON），以及用于内体逃逸的三亮氨酸（LLL）肽、用于跨越血脑屏障（BBB）和 GBM 细胞靶向的 AP-2 肽。对该 NC 进行了充分表征，并向携带颅内同基因 GL261 GBM 的小鼠静脉给药（5 次注射）。用于治疗的 NC 结构为 [P/mPEG5000(2%)/LLL(40%)/AON(alpha4beta1)(2.0%)/AP-2(2%)]。肿瘤 RNA 测序（RNA-seq）经人工智能生物信息学分析，并通过免疫组织化学对选定基因进行验证。免疫细胞群通过流式细胞术分析。结果：NC 治疗在体内抑制了 GBM 生长并显著延长了动物生存。抑制肿瘤层粘连蛋白-411 的 NC 治疗后的 RNA-seq 分析提示了抗肿瘤效应，表现为凋亡相关基因 Casp3、Ifng、Tnf、Il1a 和 Il1b 上调，胶质瘤增殖标志物及癌基因 EGFR、c-Myc、Klf4、Irf4 和 Ki-67 以及 Notch 配体 Dll3 减少。Ingenuity 通路分析的人工智能模块 AI interpret 预测激活了参与免疫应答中细胞信号传导的炎症通路、与免疫细胞黏附和迁移相关的通路以及一般免疫应答。与这些数据一致，蛋白验证显示 Ki-67 和 Dll3 减少，T 细胞标志物 CD4、炎症标志物 IFNgamma 和 TNFalpha 以及抗肿瘤 M1 巨噬细胞标志物 NOS2/iNOS 增加。有趣的是，GBM 干细胞标志物 CD133 和 nestin 的免疫染色在治疗后显著下降。流式细胞术也提示 NC 给药后免疫应答被激活。结论：我们描述了一种新型 GBM 治疗策略，即通过 NC 跨越 BBB 并靶向肿瘤微环境中关键的 ECM 和免疫成分，这些成分在很大程度上独立于胶质母细胞瘤中异质性的遗传突变。资助：NIH 基金 R01 CA284247、R01 CA188743、R01 CA206220、R01 CA209921、R01 EY013431

查看英文原文 English abstract

Introduction: Cancer microenvironment influences tumor growth, invasion, and escape from immune surveillance. Glioblastoma (GBM) extracellular matrix (ECM) especially laminins play an important role in tumor progression. We previously observed a correlation in GBM samples from 130 patients between increased expression of ECM laminin-411 (alpha4beta1gamma1) and faster tumor recurrence with decreased patient survival. Laminin-411 is a basement membrane component that can also modulate immune response through Notch signaling. We developed novel nanoconjugate (NC) blocking laminin-411 that can activate GBM local immune system after brain delivery. Methods: NC was based on non-toxic poly (beta-L-malic acid, P) with covalently attached antisense oligonucleotides (AON) against laminin-411 alpha4 and beta1 chains, as well as trileucine (LLL) peptide for endosomal escape, AP-2 peptide for blood-brain barrier (BBB) crossing and GBM cell targeting. The NC was thoroughly characterized and was intravenously administered to mice (5 injections) with intracranial syngeneic GL261 GBM. The NC selected for treatment had the structure [P/mPEG5000(2%)/LLL(40%)/AON(alpha4beta1)(2.0%)/AP-2(2%)]. Tumor RNA sequencing (RNA-seq) was analyzed by bioinformatics with artificial intelligence and validated for select genes by immunohistochemistry. Immune cell populations were analyzed by flow cytometry. Results: NC treatment in vivo suppressed GBM growth and significantly prolonged animal survival. RNA-sec analysis after treatment with NC suppressing tumor laminin-411 suggested anti-tumor effect with upregulation of apoptotic Casp3, Ifng, Tnf, Il1a, and Il1b genes and reduction of glioma proliferation markers and oncogenes EGFR, c-Myc, Klf4, Irf4, and Ki-67, as well as Notch ligand Dll3. Ingenuity pathway analysis artificial intelligence module AI interpret predicted activation of inflammatory pathways involved in cell signaling during immune responses, pathways related to adhesion and migration of immune cells, and general immune response. In agreement with these data, protein validation showed decreased Ki-67 and Dll3, and increased T cell marker CD4, inflammatory markers IFNgamma and TNFalpha, and anti-tumor M1 macrophage marker NOS2/iNOS. Interestingly, the immunostaining for markers of GBM stem cells, CD133 and nestin, was markedly decreased upon treatment. Flow cytometry also suggested activation of immune response after NC administration. Conclusion: We describe a novel GBM treatment strategy via NC crossing BBB and targeting critical ECM and immune components of tumor microenvironment that are largely independent of heterogeneous genetic mutations in glioblastoma. Support: NIH grants R01 CA284247, R01 CA188743, R01 CA206220, R01 CA209921, R01 EY013431

利益披露 Disclosure

A. V. Ljubimov, None.. R. Patil, None.. V. A. Ljubimov, None.. H. Ding, None.. Y. Wang, None.. S. Song, None.. A. A. Kramerov, None.. O. Chepurna, None.. V. Borges, None.. J. Dos Santos, None.. E. Holler, None.. J. Y. Ljubimova, None.. K. L. Black, None.

← 返回 AACR 2026 检索