Tumor-targeted delivery and cell internalization of theranostic gadolinium nanoparticles for image-guided nanoparticle-enhanced radiation therapy

用于图像引导纳米颗粒增强放射治疗的治疗诊断钆纳米颗粒的肿瘤靶向递送和细胞内化

• 批准号: 10017981
• 负责人: Wu Liu
• 金额: $ 18.27万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-13 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10017981
• 关键词: Acidity; Antibodies; Antibody Therapy; Autopsy; Biodistribution; Biological; Biological Assay; Biological Markers; Breast Cancer Model; Cell Culture Techniques; Cell Death; Cell Survival; Cell membrane; Cells; Chelating Agents; Clinical; Clinical Chemistry; Clinical Research; Comet Assay; Complement; Computer Simulation; Conformal Radiotherapy; DNA Damage; Data; Development; Diagnostic; Dose; Electrons; Endocytosis; Foundations; France; Gadolinium; Gamma-H2AX; Geometry; Goals; Hematology; Histopathology; Hour; Image; Imaging problem; In Vitro; Inductively Coupled Plasma Mass Spectrometry; Injections; Laboratory Research; Lead; Link; Magnetic Resonance Imaging; Malignant neoplasm of lung; Measures; Microscopic; Mitochondrial DNA; Modeling; Mus; Neoplasm Metastasis; Normal tissue morphology; Nuclear; Organ; Pathway interactions; Patients; Peptides; Permeability; Pharmacology; Physiological; Property; Radiation; Radiation therapy; Radiation-Sensitizing Agents; Radiosensitization; Research; Research Design; Resistance; Roentgen Rays; Signal Transduction; Solid Neoplasm; Structure; System; Techniques; Testing; Therapeutic; Time; Tissues; Toxic effect; Translating; Translations; Weight; Work; Xenograft procedure; aggressive therapy; base; biophysical model; biophysical properties; cancer cell; cell killing; cellular imaging; cellular targeting; clinical application; clinical translation; clinically relevant; experimental study; human model; image guided; imaging properties; improved; in vivo; interest; metallicity; microscopic imaging; mouse model; nanoparticle; nanoparticle delivery; neoplastic cell; novel; p53-binding protein 1; particle; pre-clinical; prevent; quantitative imaging; residence; response; simulation; targeted biomarker; targeted delivery; theranostics; translational study; treatment planning; treatment response; tumor; tumor growth; tumor microenvironment; tumor specificity; uptake

The long-term objective of this project is to overcome some of the major hurdles that prevent the clinical translation of metallic nanoparticle (NP) radiosensitization in radiation therapy (RT). Studies have shown that the passive, enhanced permeability and retention (EPR) effect itself is not sufficient to deliver the amount of intratumoral and intracellular NPs needed for in vivo radiosensitization with an affordable amount of injected NPs and the conventional NPs are cleared rapidly (~minutes) in vivo. Imaging the in vivo NP biodistribution for quantitative RT treatment planning is also an unsolved critical issue. Actively targeting and internalization into cancer cells by gadolinium (Gd) NPs conjugated to pH-Low Insertion Peptides (pHLIPs) have the potential to serve the dual purpose of enhancing uptake of NPs in tumor cells and selective, quantitative imaging by MRI. pHLIP-GdNPs can actively target solid tumors’ unavoidable acidic microenvironment, which is not present in healthy tissues. Therefore, it is superior to other biomarker targeting, such as antibody targeting, which can become nonspecific and be evaded by selection of non-expressing subclones during treatment. pHLIPs can also deliver the conjugated cell-impermeable cargoes inside the cancer cells via a strong non-endocytic pathway, critical for NP-induced short-range Auger cascade and photoelectrons to reach the vital cellular targets as proved by experiments and simulations. Complementing the rapid increasing use of MRI for RT planning and on-board treatment-guidance, pHLIP-GdNPs can also solve the imageability problem for treatment plan optimization. Our preliminary MRI data shows long tumor retention of NPs (>9 hours, possibly days) post pHLIP-GdNPs injection. Specific Aims: To provide the pre-clinical foundation for more in-depth translational and clinical studies, we aim to (i) characterize pHLIP-GdNP and evaluate its RT properties in vitro; (ii) develop a mechanistic biophysical model of radiosensitization by GdNPs to elucidate relevant biolgocial mechanisms and facilitate quantitative RT treatment planning; and (iii) investigate the in vivo radiosensitization and imaging properties of pHLIP-GdNP. Research Design: (i) Characterize pHLIP-GdNP and internalization, microscopically image cellular uptake with fluorescent tags, conduct clonogenic survival experiments in cell culture with both 250 kVp and 6 MV X-rays, generate pH-dependent cell survival curves, and examine DNA damage. (ii) Use a Monte Carlo particle track structure simulation to calculate microscopic dose enhancement induced by NPs. DNA damage will be modeled to predict sensitizer enhancement ratios and compare with experimental results. (iii) Investigate the feasibility of MR imaging to determine quantitatively in vivo NP distribution and the residence-transit time in tumor and critical organs in mouse models, the enhanced radiosensitization in vivo in mice injected with pHLIP-GdNPs compared to mice injected with untargeted GdNPs using tumor growth delay assay, and the in vivo toxicity of pHLIP-GdNPs. Impact: This project can lead to a novel theranostic agent that offers improved therapeutic ratio and imageability. This new paradigm of NP delivery and imaging can a have broad impact in image-guided NP-enhanced RT.

该项目的长期目标是克服一些阻止临床的主要障碍 辐射疗法（RT）中金属纳米颗粒（NP）放射敏的翻译。研究表明 被动，增强的渗透性和保留率（EPR）效应本身不足以交付 体内放射敏化所需的肿瘤内和细胞内NP，可承受的注射NPS 并在体内迅速清除常规的NP（〜分钟）。成像体内NP生物分布 定量RT治疗计划也是一个未解决的关键问题。积极针对和内在化 通过与pH-低插入肽（PHLIP）结合的Gadolinium（GD）NP的癌细胞具有潜力 双重目的是增强NP在肿瘤细胞中的摄取以及MRI的选择性定量成像。 phlip-gdnps可以主动瞄准实体瘤的不可避免的酸性微环境，该环境不存在 健康的组织。因此，它优于其他生物标志物靶向，例如抗体靶向，可以 在治疗过程中选择非表达亚克隆，成为非特异性的，并通过选择。 Phlips可以 还可以通过强大的非注入式途径在癌细胞内输送共轭细胞的货物 对于NP引起的短距离螺旋钻级联和光电子的至关重要 通过实验和模拟。补充MRI在RT规划和机上的迅速使用 治疗指导率，phlip-GDNP还可以解决治疗计划优化的成像性问题。我们的 初步的MRI数据显示，pHLIP-GDNPS注射后NPS（> 9小时，可能的天数）长期保留。 具体目的：为了为更深入的翻译和临床研究提供临床前的基础，我们的目标 （i）表征phlip-gdnp并在体外评估其RT特性； （ii）开发机械生物物理 GDNP的放射敏化模型以阐明相关的生物循环机制并促进定量RT 治疗计划； （iii）研究phlip-GDNP的体内放射敏化和成像特性。 研究设计：（i）表征phlip-gdnp和内在化，显微镜图像细胞摄取 荧光标签，用250 kVp和6 mV X射线进行细胞培养中的克隆生存实验， 产生pH依赖性细胞存活曲线，并检查DNA损伤。 （ii）使用蒙特卡洛粒子轨道 结构模拟以计算NP诱导的微观剂量增强。 DNA损伤将建模 预测灵敏度增强比并与实验结果进行比较。 （iii）调查 MR成像以确定体内NP分布和肿瘤中的居住时间和临界时间 小鼠模型中的器官，在注射phlip-gdnps的小鼠体内增强的放射敏化。 使用肿瘤生长延迟测定法和phlip-GDNP的体内毒性注射未靶向的GDNP的小鼠。 影响：该项目可以导致一种新型的热剂，可提供改善的治疗比和可成像性。 这种新的NP传递和成像范式可以在图像引导的NP增强RT中产生广泛的影响。