三阴性乳腺癌代谢标志物筛选、鉴定及蛋白靶向探针可视化研究

Accumulate evidence suggests that hypoxia is associated with initiation, invasiveness, and metastasis of triple negative breast cancer (TNBC), but limited markers related to this process have been identified. Oxygen is a major determinant for cell metabolism, and the greatest amount (about 85-90%) of oxygen is consumed by mitochondria in cells to allow oxidative phosphorylation for ATP generation. However, few specific marker-enzymes involved in mitochondrial metabolism under tumor hypoxic conditions have been discovered. In this proposal, we will investigate the roles of hypoxia in TNBC progression and the dynamic changes of ferritin receptor, TfR 1, which has been newly identified as a tumor marker. Meanwhile, we will integrate metabolomics (metabolites), transcriptomics (mRNA) and proteomics (proteins) data to screen and/or find the specific marker-enzymes of mitochondrial metabolism and further explore how the marker-enzymes expression is regulated by hypoxia and their roles in TNBC procession. Based on these mechanisms, human ferritin-based nanocages (FTn) will be designed as carriers: (a) to develop a non-antibody based technique for TfR 1 detection in TNBC tumor tissues; (b) to design a FTn-based probe for tumor hypoxia targeting and specific tumor mitochondrial metabolic marker-enzymes response to accomplish tumor diagnosis and therapy simultaneously. This proposal not only meets the requirements of the related Major Program of the NSFC, but also plays important role in understanding of tumor pathological progression and development of the related theranostic techniques.

低氧在三阴性乳腺癌（TNBC）的发生、发展及组织转移中起着重要的作用，目前对于过程中低氧相关标志物分子所知很少；氧气是决定细胞代谢的重要分子，而线粒体作为对氧气最为依赖的细胞器，在肿瘤低氧条件下是否存在特异性酶标志物还不清楚。本项目拟探索低氧在TNBC肿瘤演进中作用及肿瘤标志物分子转铁蛋白受体1（TfR 1）动态变化规律；拟利用组学等技术筛选和/或鉴定线粒体代谢标志物酶，理解低氧对其调控机理和在TNBC肿瘤演进过程中作用。基于这些机理的解析，通过基因工程化人源铁蛋白纳米笼载体作为核心，（1）发展识别肿瘤标志物TfR 1的非抗体技术，探索其在组织病理学可视化体外诊断的应用；（2）开发低氧靶向、线粒体代谢标志物酶特异性响应的蛋白纳米探针，用于体内可视化成像肿瘤和诊疗一体化。本项目不仅紧扣相关重大研究计划的要求，而且对于理解TNBC的病理过程和开发针对性的诊疗技术具有重要的借鉴意义。

肿瘤低氧对于肿瘤的发生和演进起着关键性作用，发展针对低氧有关的可视化技术和治疗策略至关重要。项目以可结合TfR1的铁蛋白作为切入点，开发了基于铁蛋白的探针和药物，用于肿瘤（特别是三阴性乳腺癌）及微环境的可视化和治疗，主要包括三方面内容：（i）证实了TfR 1的表达与肿瘤（包括三阴性乳腺癌）低氧正相关性，确定了基于铁蛋白作为载体靶向肿瘤低氧的可行性，设计和发展了铁蛋白组装体、铁蛋白蛋白冠等为基础的多种肿瘤靶向和治疗策略。（ii）证实了代谢标志物酶P450与低氧有关，以P450裂解的化学键为基础开发了探针和前药，并以铁蛋白纳米酶模拟P450活性开发前药递送系统，用于三阴性乳腺癌的诊疗。（iii）利用铁蛋白纳米探针，结合人工智能图像分割和识别技术，开发可量化肿瘤血管渗透性的可视化新技术，研究肿瘤（包括三阴性乳腺癌）血管渗透的异质性，揭示肿瘤渗透机制，并提出差异化设计纳米药物的新策略。这些研究为开发肿瘤的可视化技术提供了选择，也为理解肿瘤的异质性及开发个性化治疗手段提供了参考。在项目支持下，已发表或已接受SCI论文14篇（含有基金号），包括1篇Nature Nanotech、6篇Advanced Mater、1篇Science Adv、1篇Advanced Funct Mater、1篇Small、2篇Theranostics、1篇ACS Appl Mater Interfaces、1篇Bioconjugate Chem。获批国家发明专利2项，提交发明专利1项。