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

• 批准号: MR/X01116X/1
• 负责人: Adam Perriman
• 金额: $ 73.4万
• 依托单位: University of Bristol
• 依托单位国家: 英国
• 项目类别: Fellowship
• 财政年份: 2023
• 资助国家: 英国
• 起止时间: 2023 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=MR%2FX01116X%2F1
• 关键词: Supercharged; protein; surfactant; bioconjugates; next

Artificial membrane binding proteins (AMBPs) have the potential to impact on the efficacy of adoptive cell therapies, as the introduction of exogenous proteins to provide additional functionality to therapeutic cells could be highly advantageous for site-directed tissue repair. The methodology is centred on the rational design of modular bifunctional supercharged protein-polymer surfactant complexes that spontaneously insert into the plasma membrane of stem cells to impart oxygen delivery and chemotropic homing properties. This could address many of the challenges in cell therapies, including the lack of viable cell engraftment, which results in limited functional integration. Several cell therapy studies have shown that intravenous or intra-arterial infusion of stem cells leads to accumulation in tissue sinks, such as the lungs and spleen. These off-target effects reduce the efficiency of systemic delivery and increase the likelihood of producing lethal microemboli. Accordingly, this research programme describes the rational design of a new class of bifunctional AMBPs with responsive oxygen delivery and 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 circumvents the need for covalent cell-surface chemistry, and offers a high degree of flexibility, as the approach can be applied to a wide range of proteins for use on potentially any cell type. The AMBP methodology pioneered by the PI involves the rational design of an AMBP in two key steps: (i) supercharging the AMBP anchor to amplify the positive surface charge density (ii) electrostatic grafting of polymer surfactant chains to the cationic sites on the membrane anchor. The resulting polymer surfactant corona allows the cell membrane affinity to be systematically tuned to facilitate spontaneous insertion of the AMBP into the cell membrane, whilst retaining the native function of the cell-bound protein. The PI has successfully applied this methodology to the oxygen-binding protein myoglobin, as well as supercharged green fluorescent protein fused to the fibronectin binding domain of a bacterial adhesin motif CshA. Here, the AMBPs rapidly inserted into the membranes of adult bone-marrow derived hMSCs and provided either responsive oxygen delivery or chemotactic stem cell homing to cardiac tissue. The research programme has a strong (but not exclusive) focus on developing AMBPs for cardiac stem cell therapies, as cardiovascular disease (CVD) is the leading cause of death globally (2021 World Health Organisation estimate is 17.9 million deaths p.a.). Here, the conceptual advance is centred on developing humanised bifunctional AMBP chimeras that can responsively deliver oxygen to the cells via a supercharged myoglobin anchor module (to improve cell viability in hypoxic in vivo environments), fused to a module that provides cardiac extracellular matrix targeting. Accordingly, the ability to display multiple copies of these AMBPs, which can deliver oxygen and have been evolutionarily optimised to recognise and bind specific molecular targets in the cardiac endothelium, has the potential to advance cardiac cell therapy.The AMBP platform is likely to have clinical impact beyond cell therapies for CVD, as it could be readily applied to other cell types and vesicles (e.g., monocytes, natural killer cells, exosomes, or lipid nanoparticles) and involve other homing protein- or peptide-based molecules (e.g., integrins, nanobodies, or other bacterial adhesins). The research programme describes a scientific methodology that combines both in-house techniques for biophysics, synthetic biology 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 scientists and industrial partners to aid with medical translation.

人工膜结合蛋白（ambp）有可能影响过继细胞治疗的疗效，因为引入外源蛋白为治疗细胞提供额外的功能可能对位点定向组织修复非常有利。该方法的核心是模块化双功能增压蛋白质-聚合物表面活性剂复合物的合理设计，这些复合物可以自发地插入干细胞的质膜中，以赋予氧气输送和趋化归巢特性。这可以解决细胞治疗中的许多挑战，包括缺乏活细胞植入，导致有限的功能整合。几项细胞治疗研究表明，静脉或动脉内输注干细胞会导致组织池积聚，如肺和脾。这些脱靶效应降低了全身给药的效率，增加了产生致命微栓塞的可能性。因此，本研究计划描述了一类新的双功能ambp的合理设计，这些ambp具有响应性氧输送和化学营养归巢特性，将有助于克服这些限制，并在疾病和再生医学的细胞治疗中具有深远的意义。这种新方法绕过了共价细胞表面化学的需要，并提供了高度的灵活性，因为这种方法可以应用于广泛的蛋白质，用于任何细胞类型。由PI首创的AMBP方法涉及合理设计AMBP的两个关键步骤：(i)对AMBP锚进行增压以放大正电荷表面密度；（ii）将聚合物表面活性剂链静电接枝到膜锚上的阳离子位点。由此产生的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AMBP自发插入细胞膜，同时保留细胞结合蛋白的天然功能。PI已经成功地将这种方法应用于氧结合蛋白肌红蛋白，以及与细菌粘附素基模CshA的纤维连接蛋白结合域融合的增压绿色荧光蛋白。在这里，ambp迅速插入成人骨髓来源的hMSCs的膜，并提供响应氧输送或趋化干细胞归巢到心脏组织。由于心血管疾病（CVD）是全球死亡的主要原因（2021年世界卫生组织估计每年有1790万人死亡），该研究计划强烈（但不是唯一）关注于开发用于心脏干细胞治疗的ambp。在这里，概念上的进展集中在开发人源化双功能AMBP嵌合体上，该嵌合体可以通过增压肌红蛋白锚定模块（以提高体内缺氧环境下的细胞活力）响应性地向细胞输送氧气，并融合到提供心脏细胞外基质靶向的模块中。因此，显示这些ambp的多个拷贝的能力具有推进心脏细胞治疗的潜力，这些ambp可以输送氧气，并经过进化优化以识别和结合心脏内皮中的特定分子靶标。AMBP平台可能会在CVD的细胞治疗之外产生临床影响，因为它可以很容易地应用于其他细胞类型和囊泡（例如，单核细胞、自然杀伤细胞、外泌体或脂质纳米颗粒），并涉及其他基于归巢蛋白或肽的分子（例如，整合素、纳米体或其他细菌粘附素）。该研究计划描述了一种科学方法，它结合了生物物理学、合成生物学和再生医学的内部技术，以及大型设施中可用的尖端技术。由于该项目有很强的医学重点，申请人已经聘请了临床科学家和工业合作伙伴来协助医学翻译。

项目成果

Technological advances in the use of viral and non-viral vectors for delivering genetic and non-genetic cargos for cancer therapy.

• DOI: 10.1007/s13346-023-01362-3
• 发表时间: 2023-11
• 期刊: DRUG DELIVERY AND TRANSLATIONAL RESEARCH
• 影响因子: 5.4
• 作者: Dogbey, Dennis Makafui;Torres, Valeria Esperanza Sandoval;Fajemisin, Emmanuel;Mpondo, Liyabona;Ngwenya, Takunda;Akinrinmade, Olusiji Alex;Perriman, Adam W.;Barth, Stefan
• 通讯作者: Barth, Stefan

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.

Bienzymatic Generation of Interpenetrating Polymer Networked Engineered Living Materials with Shape Changing Properties

• DOI: 10.1002/admt.202300626
• 发表时间: 2023-07
• 期刊: Advanced Materials Technologies
• 影响因子: 6.8
• 作者: R. Klemperer;Mark R. Shannon;J. L. Ross Anderson;A. Perriman

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