Structural precision meets bio-functionality: Virus-like delivery systems for genetic material

遗传物质的靶向递送在治疗应用中变得越来越重要。病毒是非常有效的基因传递系统，但表现出货物大小的限制，并在人体内引起显着的免疫反应。在此，我们提出了聚合载体的发展，形成病毒样颗粒后，与遗传物质凝聚。通过使用分段瓶刷共聚物和先进的生物缀合策略，我们将开发具有核心隔室（包含货物），模仿病毒衣壳的保护壳和蛋白质冠的聚合物，从而实现与细胞的靶向相互作用。由于其空间受限的性质，瓶刷共聚物理想地适合于模拟病毒的基于蛋白质的超分子结构单元。通过对聚合物结构和组成的系统研究，将调整聚合物复合物的大小、形状、稳定性和生物相互作用概况以实现有效递送。将开发一种新的糖基化策略，以单步和位点特异性的方式将蛋白质配体偶联到递送系统。这将进一步微调递送系统的相互作用谱，并实现与特定细胞类型的靶向相互作用，癌细胞所开发的复合物的功能特征在于它们的作用模式，在细胞培养模型中使用报告基因和自杀基因，并在反馈回路中迭代优化。蛋白质将通过基因工程进行修饰，以实现应变炔的基于糖基转移酶的偶联，这反过来又可以通过应变促进的点击化学偶联到聚合复合物。该项目将通过高效和直接的合成方法获得先进和结构复杂的基因递送系统。此外，病毒样复合物的设计和它们的结构属性关系的基本知识将产生使该平台适应各种目的。

The targeted delivery of genetic material is becoming increasingly important in therapeutic applications. Viruses are highly effective gene delivery systems but show restrictions in cargo size and cause significant immune responses in the human body. We herein propose the development of polymeric vectors that form virus-like particles upon condensation with genetic material. By using segmented bottle brush copolymers and advanced bioconjugation strategies, we will develop polyplexes featuring a core compartment (containing the cargo), a protective shell mimicking the viral capsid, and a protein corona enabling targeted interactions with cells. Due to their sterically confined nature bottle brush copolymers are ideally suited to mimic the protein-based supramolecular building blocks of a virus. By systematic investigation of polymer structure and composition, the size, shape, stability, and biological interaction profile of polyplexes will be tuned to enable efficient delivery. A new glycosylation strategy will be developed to couple protein ligands to the delivery system in a single-step and site-specific way. This will further fine-tune the interaction profile of the delivery system and enable targeted interactions with specific cell types, e.g., cancer cells. The developed polyplexes are functionally characterized in their mode of action using a reporter gene and a suicide gene in cell culture models and iteratively optimized in feedback loops. Proteins will be modified via genetic engineering to enable glycosyltransferase-based coupling of strained alkyne, which in turn can be coupled to polyplexes via strain promoted click chemistry. This project will enable access to advanced and structurally complex gene delivery systems via efficient and straight forward synthetic methods. In addition, fundamental knowledge about the design of virus-like polyplexes and their structure property relationships will be generated enabling adaption of this platform to various purposes.