Hepsin cleavable nanocapsules for precision medicine in prostate carcinoma (HEPSIN-NANOLYSIS

丝氨酸蛋白酶hepsin在90%的前列腺癌病例中过表达，表现出细胞周围蛋白水解活性，并促进小鼠肿瘤进展和转移。由于这些特性，hepsin是前列腺癌诊断和治疗的一个有吸引力的靶分子。我们打算利用hepsin的酶活性来进行抗肿瘤药物的位点特异性释放。一种新的，基于肽的纳米药物载体系统将被开发和评估。纳米载体将被设计用于在前列腺癌细胞的表面选择性打开，从而能够在显著减少副作用的情况下进行特异性和有效的治疗干预。LY 294002是一种完全表征的PI 3-激酶/AKT介导的信号传导的拮抗剂，在前列腺癌细胞系中显示出有效的抗肿瘤活性，单独或与促凋亡和癌症特异性细胞因子TRAIL（肿瘤坏死因子相关凋亡诱导配体）组合。通过将这些药物包封在待开发的纳米载体系统中，我们的目标是显著提高生物利用度和功效，并将体内副作用降至最低。将使用鸡绒毛尿囊膜（CAM）模型中具有hepsin诱导表达的实验性前列腺癌异种移植物对该概念进行生物学评价。将使用反相细乳液方法制备胶囊。将优化共聚单体和合成策略。肝蛋白酶特异性靶序列的选择将通过筛选基于珠粒的组合肽文库来实现。将沿着对表面性质的反复调整，分析生物介质中肽基胶囊的聚集和吸附特性，最终得到纳米载体，其i）是完全可生物降解的; ii）在肿瘤中有效地积累，但不在血管或器官中; iii）显示可用于蛋白水解的表面; iv）可被hepsin特异性降解;和v）在暴露于具有酶活性的hepsin蛋白酶时快速释放其货物。该跨学科项目将为即将到来的旨在系统开发和评估用于癌症治疗的酶可裂解纳米载体的方法提供基础。除了建立基于文库的筛选技术和纳米载体合成和负载的通用策略外，高度标准化的体外模型以及CAM异种移植模型中的体内分析将在临床前研究中提供可靠的数据，这将在避免动物试验的情况下实现。

The serine protease hepsin is overexpressed in 90% of prostate cancer cases, exhibits pericellular proteolytic activity, and promotes tumor progression and metastasis in mice. Due to these features, hepsin is an attractive target molecule for both diagnosis and therapy of prostate cancer. We intend to use the enzymatic activity of hepsin for the site-specific release of antitumour drugs. A novel, peptide-based nanoscale drug carrier system will be developed and evaluated. The nanocarriers will be designed for selective opening at the surface of prostate cancer cells, thereby enabling specific and efficient therapeutic intervention under considerable reduction of side effects.LY294002, a thoroughly characterized antagonist of PI3-Kinase / AKT-mediated signalling, reveals potent anti-tumor activity in prostate cancer cell lines, either alone or combined with the pro-apoptotic and cancer-specific cytokine TRAIL (tumor necrosis factor related apoptosis-inducing ligand). By encapsulating these drugs in the nanocarrier system to be developed, we aim at a significant improvement of bioavailabilty and efficacy and a minimization of side effects in vivo. The concept will be biologically evaluated using experimental prostate cancer xenografts featuring inducible expression of hepsin in the chicken chorioallantoic membrane (CAM) model.The capsules will be prepared using the inverse miniemulsion method. Co-monomers and synthesis strategies will be optimized. The selection of a hepsin-specific target sequence will be achieved by screening of a bead-based combinatorial peptide library. Aggregation- and adsorption characteristics of peptide-based capsules in biological media will be analysed along with iterative adaptations of surface properties, ultimately leading to nanocarriers which i) are completely biodegradable; ii) efficiently accumulate in tumors, but not in vessels or organs; iii) reveal a surface which is accessible for proteolysis; iv) are specifically degradable by hepsin; and v) rapidly release their cargo upon exposure to enzymatically active hepsin protease. The interdisciplinary project will provide a groundwork for upcoming approaches aiming at the systematic development and evaluation of enzymatically cleavable nanocarriers for cancer therapy. In addition to the establishment of a library-based screening technology and a universal strategy for nanocarrier synthesis and loading, highly standardized in vitro models as well as in vivo analyses in the CAM xenograft model will provide reliable data in preclinical research, which will be achieved under avoidance of animal trials.