Optimal cell factories for membrane protein production

用于膜蛋白生产的最佳细胞工厂

• 批准号: BB/Y007603/1
• 负责人: Alexander Darlington
• 金额: $ 186.27万
• 依托单位: University of Warwick
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2024
• 资助国家: 英国
• 起止时间: 2024 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FY007603%2F1
• 关键词: Optimal; cell; factories; membrane; protein

SOCIETAL IMPORTANCE. Membrane proteins are essential for life, medicine and industrial biotechnology and yet remain understudied due to their (1) low abundance and (2) physical properties which makes isolation difficult. However, 20-30% of all proteins are membrane proteins with biological functions underpinning signal transduction, nutrient import/export, and cell-to-cell communication. Membrane proteins are potent drug and vaccine targets. Currently, the top ten selling medicines worldwide, and in total US$ 180 billion worth of drug sales, target membrane proteins. Multiple vaccines, including those for COVID-19, Hepatitis B and pertussis, target virus surface or bacterial membrane proteins. Non-pharmaceutical markets include biopesticides (worth US$ 3 billion) and antifungals (worth US$ 19 billion). Artificial expression of membrane proteins is frequently used in pharmacology to identify drug molecules, which may inhibit or enhance protein function. However, engineering high levels of functional membrane protein production remains challenging, limiting the efficiency of drug screening, reducing identification of new drugs and limiting transformative potential. Production of membrane proteins enables progress in promoting human health and provides bio-inspired solutions to pressing societal challenges caused by climate change; however, obtaining high yields of functional protein constitutes a fundamental roadblock to progress which this proposal addresses.SCIENTIFIC CHALLENGE. Despite the importance of membrane proteins in these key industries, their production remains challenging with production of each protein requiring trial and error methods - in some cases requiring screening of 100s of different combinations of experimental factors. This is in part due to the complex process of membrane protein production which results in multiple stresses on cellular production platforms (significantly reducing cell growth and bioprocess productivity). The cell membrane's lipid composition significantly impacts protein function - potentially rendering any proteins produced non-functional due to lack of lipid factors. Therefore, scientists must test multiple different cell types in an expensive and laborious process. This bespoke platform development limits production capacity of functional membrane proteins and, in doing so limits drug screening and development.PROJECT OUTLINE. We address these challenges by first carrying out a detailed systematic characterization of the cellular processes which limit membrane protein production. This will be used to develop a predictive computer aided design tool to enable designs which facilitate both production and cell growth. We will use these tools to design and implement autonomous feedback control strategies that enable living cells to self-regulate protein production in response to stress and therefore maximise production and yield. These systems have the potential to enable increased yields in a 'hands off manner'. We will engineer new cellular regulatory systems capable of tuning the cell's lipid production rate so that the cell membranes' composition and physical properties can be tuned to meet those needed for optimal membrane protein function. We will develop computer aided design tools which enable the prediction of optimal membrane composition. We will demonstrate the function of our new cellular systems to produce high value biomedical membrane proteins and demonstrate their production performance at scale in industrially relevant conditions.

社会重要性。膜蛋白对于生命、医学和工业生物技术是必不可少的，但由于它们的（1）低丰度和（2）使得分离困难的物理性质而仍然研究不足。然而，所有蛋白质的20-30%是膜蛋白，其生物学功能支持信号转导、营养输入/输出和细胞间通讯。膜蛋白是有效的药物和疫苗靶点。目前，全世界销售额最高的10种药物，总销售额为1800亿美元，靶向膜蛋白。多种疫苗，包括针对COVID-19、乙型肝炎B和百日咳的疫苗，针对病毒表面或细菌膜蛋白。非药品市场包括生物杀虫剂（价值30亿美元）和抗真菌剂（价值190亿美元）。膜蛋白的人工表达常用于药理学中以鉴定可抑制或增强蛋白质功能的药物分子。然而，工程化高水平的功能性膜蛋白生产仍然具有挑战性，限制了药物筛选的效率，减少了新药的鉴定，并限制了转化潜力。膜蛋白的生产可以促进人类健康，并为气候变化带来的紧迫社会挑战提供生物启发的解决方案;然而，获得高产量的功能蛋白构成了本提案所解决的进步的根本障碍。科学挑战尽管膜蛋白在这些关键行业中的重要性，但它们的生产仍然具有挑战性，每种蛋白质的生产都需要试错法-在某些情况下需要筛选数百种不同的实验因素组合。这部分是由于膜蛋白生产的复杂过程，其导致细胞生产平台上的多重应力（显著降低细胞生长和生物过程生产率）。细胞膜的脂质组成显著影响蛋白质功能-由于缺乏脂质因子，可能使任何产生的蛋白质无功能。因此，科学家们必须在昂贵而费力的过程中测试多种不同的细胞类型。这种定制的平台开发限制了功能性膜蛋白的生产能力，从而限制了药物筛选和开发。我们首先对限制膜蛋白产生的细胞过程进行详细的系统表征，以应对这些挑战。这将用于开发预测性计算机辅助设计工具，以实现有利于生产和细胞生长的设计。我们将使用这些工具来设计和实施自主反馈控制策略，使活细胞能够自我调节蛋白质生产以应对压力，从而最大限度地提高产量和产量。这些系统具有以“不干涉的方式”提高产量的潜力。我们将设计新的细胞调节系统，能够调节细胞的脂质产生速率，从而可以调节细胞膜的组成和物理特性，以满足最佳膜蛋白功能所需的条件。我们将开发计算机辅助设计工具，使最佳膜组成的预测。我们将展示我们的新细胞系统的功能，以生产高价值的生物医学膜蛋白，并在工业相关条件下展示其大规模生产性能。