Supercharged protein-surfactant bioconjugates for next-generation cell therapies

用于下一代细胞疗法的增压蛋白质-表面活性剂生物缀合物

基本信息

批准号：
MR/S016430/1
负责人：
Adam Perriman
金额：
$ 119.13万
依托单位：
University of Bristol
依托单位国家：
英国
项目类别：
Fellowship
财政年份：
2019
资助国家：
英国
起止时间：
2019 至无数据
项目状态：
已结题

来源：
https://gtr.ukri.org/projects?ref=MR%2FS016430%2F1
关键词：
Supercharged protein surfactant bioconjugates next

项目摘要

Cell membrane engineering is a rapidly emerging field with significant potential to impact on cell therapies, as the introduction of exogenous proteins into the cell membrane is highly advantageous for in vivo site-directed tissue repair. This is because there exist a number of barriers to the widespread uptake of cell therapies, including the lack of engraftment of transplanted cells, which results in limited functional integration. Several cell therapy studies have shown that intravenous or intra-arterial infusion of stem cells leads to undesirable accumulation in the lungs, which reduces the efficiency of systemic delivery and increases the likelihood of producing lethal microemboli. Even when implanted directly into the organ of interest, the number of cells required for therapeutic benefit can be prohibitively high. Accordingly, this research programme describes the rational design of a new class of artificial membrane binding proteins with chemotrophic homing properties that will help overcome these limitations and have far-reaching implications in cell therapies for disease and regenerative medicine.The new approach involves cell functionalisation with protein-polymer surfactant bioconjugates, which circumvents the need for direct cell-surface chemistry, and offers a high degree of flexibility, as the approach can be readily applied to a diverse range of proteins for use on potentially any cell type. The synthetic methodology recently pioneered by the PI involves the re-engineering of a protein surface in two key steps: (i) amplification of the positive charge density on the protein surface (supercharging) (ii) electrostatic coupling of anionic polymer surfactant chains to the cationic sites displayed on the protein surface. Significantly, by constructing this polymer surfactant corona, the cell membrane affinity can be systematically tuned to facilitate spontaneous insertion of the bioconjugate into a stem cell membrane, whilst retaining the native function of the protein linked to the cell surface. The PI recently applied this methodology to the oxygen-binding protein myoglobin and demonstrated that the bioconjugates rapidly inserted into the cytoplasmic membranes of adult bone-marrow derived human mesenchymal stem cells (hMSCs). The research programme has a strong (but not exclusive) focus on developing the bioconjugates for cell homing for cardiac stem cell therapy, as cardiovascular disease (CVD) is the leading cause of death globally (2015 WHO estimate is 18 million deaths p.a.). Here, the conceptual advance is centred on hijacking the homing properties of infectious bacteria by immobilising the fibronectin (Fn) binding domain of the bacterial adhesion protein CshA on the cytoplasmic membranes of stem cells. The CshA-Fn interaction has been proposed to be responsible for directing and immobilising S. gordonii colonisation at the cardiac endothelium, promoting the onset of infective endocarditis. Accordingly, the ability to display multiple copies of this binding motif, which has been evolutionarily optimised to recognise and bind specific molecular targets in the cardiac endothelium, on the membrane of stem cells has the potential to advance cardiac cell therapy.Significantly, the cell membrane modification platform has the potential to have clinical impact beyond cell therapies for CVD, as it could be readily applied to other cell types and vesicles (e.g., monocytes, blood outgrowth endothelial cells, natural killer cells or exosomes) and involve other homing protein- or peptide-based molecules (e.g., integrins, antibodies, or other bacterial adhesins). The research programme describes a scientific approach that combines in-house techniques for biophysics and regenerative medicine, as well as cutting-edge techniques available at large-scale facilities. As there is a strong medical focus within the programme, the applicant has engaged clinical scientist partners to aid with medical translation.

细胞膜工程是一个快速新兴的领域，具有对细胞疗法影响的重要潜力，因为将外源蛋白引入细胞膜对于体内定位的组织修复非常有利。这是因为存在广泛摄取细胞疗法的许多障碍，包括缺乏移植细胞的植入，从而导致功能积分有限。几项细胞疗法研究表明，干细胞的静脉内或动脉内输注导致肺部的不良积累，这降低了全身性递送的效率并增加了产生致命微栓塞的可能性。即使直接植入感兴趣的器官，治疗益处所需的细胞数量也可能很高。 Accordingly, this research programme describes the rational design of a new class of artificial membrane binding proteins with chemotrophic homing properties that will help overcome these limitations and have far-reaching implications in cell therapies for disease and regenerative medicine.The new approach involves cell functionalisation with protein-polymer surfactant bioconjugates, which circumvents the need for direct cell-surface chemistry, and offers a high degree of灵活性，因为该方法可以很容易地应用于各种蛋白质范围，以用于潜在的任何细胞类型。 PI最近先开拓的合成方法涉及在两个关键步骤中重新设计蛋白质表面：（i）在蛋白质表面上的正电荷密度扩增（增压）（II）在蛋白质表面显示的阳离子表面上的阴离子聚合物表面活性剂链静电偶联。值得注意的是，通过构建该聚合物表面活性剂电晕，可以系统地调节细胞膜亲和力，以促进生物缀合物自发插入到干细胞膜中，同时保留与细胞表面链接的蛋白质的天然功能。 PI最近将这种方法应用于氧结合蛋白肌红蛋白，并证明了生物缀合物迅速插入成人骨row的人骨髓中质质细胞的细胞质膜（HMSC）。该研究计划强烈（但不是排他性的）专注于开发用于心脏干细胞疗法的细胞归巢的生物缀合物，因为心血管疾病（CVD）是全球死亡的主要原因（2015年估计为1800万死亡P.A.）。在这里，概念上的前进是通过固定细菌粘附蛋白CSHA的纤连蛋白（FN）结合结构域在干细胞的细胞质膜上劫持感染性细菌的归巢特性。已提出CSHA-FN相互作用负责指导和固定在心脏内皮上的Gordonii定植，从而促进感染性心内膜炎的发作。因此，在干细胞的膜上显示该结合基序的多个结合基序的多个副本的能力在进化上进行了优化，以识别和结合心脏内皮中特定的分子靶标的能力，有可能提高心脏细胞治疗的可能性，从而有明显地提高心脏膜的修饰平台，可以在细胞中使用其他细胞的临床影响。单核细胞，血液生长内皮细胞，天然杀伤细胞或外泌体），涉及其他基于蛋白质或肽的分子（例如整联蛋白，抗体或其他细菌粘附素）。该研究计划描述了一种科学方法，该方法结合了内部的生物物理学和再生医学技术，以及大型设施可用的尖端技术。由于该计划中有强烈的医疗重点，因此申请人与临床科学家合作伙伴聘请了医学翻译。

项目成果

期刊论文数量（10）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Development of a Novel Hierarchically Biofabricated Blood Vessel Mimic Decorated with Three Vascular Cell Populations for the Reconstruction of Small-Diameter Arteries

DOI：
10.1002/adfm.202300621
发表时间：
2023-11-03
期刊：
ADVANCED FUNCTIONAL MATERIALS
影响因子：
19
作者：
Carrabba，Michele;Fagnano，Marco;Madeddu，Paolo
通讯作者：
Madeddu，Paolo

Fabrication of New Hybrid Scaffolds for in vivo Perivascular Application to Treat Limb Ischemia.

DOI：
10.3389/fcvm.2020.598890
发表时间：
2020
期刊：
Frontiers in cardiovascular medicine
影响因子：
3.6
作者：
Carrabba M;Jover E;Fagnano M;Thomas AC;Avolio E;Richardson T;Carter B;Vozzi G;Perriman AW;Madeddu P
通讯作者：
Madeddu P

Modular Bioorthogonal Lipid Nanoparticle Modification Platforms for Cardiac Homing.

DOI：
10.1021/jacs.3c07811
发表时间：
2023-10-18
期刊：
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY
影响因子：
15
作者：
Cruz-Samperio, Raquel;Hicks, Corrigan L.;Scott, Aaron;Gispert Contamina, Ignacio;Elani, Yuval;Richardson, Rebecca J.;Perriman, Adam W.
通讯作者：
Perriman, Adam W.

Cell augmentation strategies for cardiac stem cell therapies.