A Wholly Protein-based Self-assembly Nanoplatform for TNBC-specific Combination Therapy

摘要 三阴性乳腺癌（TNBC）是一种独特类型的乳腺癌，不表达或 过表达雌激素受体（ER）、孕激素受体（PR）和HER 2。在过去的几十年里， TNBC的标准治疗仍然是高毒性化疗，在更有效的治疗方面几乎没有进展。 为了解决开发特异性针对TNBC的靶向疗法的迫切需求，我们开发了一种完全 一种基于蛋白质的纳米平台，称为ProNano，由两种重组蛋白组成，包括第一种和第二种。 弹性蛋白样多肽（ELP）纳米核，其在表面上展示多个钙调蛋白结合短肽， 第二种是与高度稳定和模块化的蛋白质遗传融合的重组钙调蛋白 具有肿瘤归巢或治疗特征的结构域。ELP纳米核与TNBC的对接- 靶向模块与具有治疗特征的功能模块一起产生双功能ProNanos， 可用于TNBC联合治疗的治疗剂的肿瘤归巢递送。三 该项目将追求具体目标。第一个具体目标是开发一个ProNano平台， 基于TNBC细胞的表面抗原表达谱，可调节靶向TNBC细胞。第二个具体目标是 开发一种双功能ProNano平台，允许肿瘤特异性阻断异常Wnt信号传导， TNBC的治疗第三个具体目标是开发一种双功能ProNano平台，允许肿瘤- 特异性抑制ENPP 1催化的细胞外cGAMP水解，用于联合免疫治疗 TNBC。在这个项目中开发的双功能ProNano平台有几个主要优点， 传统的纳米平台，包括所有蛋白质组分，每个都可以被精确地和遗传地工程化， 肿瘤归巢和/或治疗部分以所需比例的定向和自组装引入 不需要任何化学偶联，并使用高度稳定和易于表达的模块化多肽 没有或低免疫原性。虽然我们专注于TNBC的靶向治疗，但ProNano平台 在这个项目中开发的，可以很容易地适应于解决其他类型的癌症， 瞄准模块

Abstract Triple-negative breast cancer (TNBC) is a unique type of breast cancer that does not express or overexpress estrogen receptor (ER), progesterone receptor (PR), and HER2. During the past several decades, the standard care of TNBC remains the highly toxic chemotherapy with little progress in more effective treatments. To address the urgent unmet need to develop targeted therapies specific for TNBC, we developed a totally protein-based nanoplatform called ProNano that is composed of two recombinant proteins, including the first an elastin-like polypeptide (ELP) nanocore that displays multiple calmodulin-binding short peptides on the surface, and the second a recombinant calmodulin protein genetically fused with a highly stable and modular protein domain possessing either tumor homing or therapeutic features. The docking of the ELP nanocore with a TNBC- targeting module together with a functional module with therapeutic feature results in bifunctional ProNanos that can be used for tumor-homing delivery of therapeutic agents for the combination therapy for TNBC. Three specific aims will be pursued in this project. The first specific aim is to develop a ProNano platform that allows tunable targeting of TNBC cells based on their surface antigen expression profiles. The second specific aim is to develop a bifunctional ProNano platform that allows tumor-specific blockade of the aberrant Wnt signaling for the treatment of TNBC. The third specific aim is to develop a bifunctional ProNano platform that allows tumor- specific inhibition of the ENPP1-catalyzed hydrolysis of extracellular cGAMP for combination immunotherapy of TNBC. The bifunctional ProNano platforms developed in this project have several major advantages over conventional nanoplatforms, including all protein components each can be precisely and genetically engineered, oriented and self-assembled introduction of both tumor homing and/or therapeutic moieties at desired ratios without need of any chemical conjugation, and use of highly stable and easily expressed modular polypeptides with no or low immunogenicity. Although we focus on the targeted treatment for TNBC, the ProNano platforms developed in this project can be easily adapted to address other cancer types simply by changing the tumor targeting module.