Investigation and manipulation of mTOR cellular signalling to generate novel CHO host cells with high growth and productivity characteristics

研究和操作 mTOR 细胞信号传导以产生具有高生长和生产力特征的新型 CHO 宿主细胞

基本信息

  • 批准号:
    BB/J006408/1
  • 负责人:
  • 金额:
    $ 41.31万
  • 依托单位:
  • 依托单位国家:
    英国
  • 项目类别:
    Research Grant
  • 财政年份:
    2012
  • 资助国家:
    英国
  • 起止时间:
    2012 至 无数据
  • 项目状态:
    已结题

项目摘要

Small molecule drugs (e.g. antibiotics) have traditionally been the mainstay of treatments and therapies in man, however in the last 10-20 years protein based drugs (e.g. herceptin, often used to treat breast cancer) have developed such that these now constitute a significant section of the pharmaceutical market. There are several categories of protein based drugs, one of which, monoclonal antibodies, constitutes the largest number of protein molecules in a class either in use or in clinical trials. Many protein based drugs are challenging to produce because they (a) require particular helper proteins to fold and assemble into their final active state and (b) are decorated on their surfaces by sugars and other molecules that are essential to their bioactivity. Due to the high precision required to produce such biotherapeutics, such 'recombinant' protein-based drugs for the treatment of diseases are usually produced by cells kept in culture under defined conditions. One problem with this is that the cells we use to make proteins for therapeutic uses are not as efficient as we would like them to be. As a consequence, we may not be able to produce enough of these drugs and/or the cost of producing them may be too high for health care providers. This proposal therefore sets out to address a key area that underpins recombinant protein synthesis from mammalian cells. It aims to provide understanding of how a global regulator of protein synthesis, mTOR (mammalian target of rapamycin), contributes to recombinant protein synthesis and devise new ways to manipulate this process to enhance recombinant protein yields.Protein synthesis is the process by which the information in the genetic material in the cell, DNA is converted via an intermediary messenger, termed mRNA, into proteins such as antibodies. Protein synthesis takes part on ribosomes and mTOR signalling also effects ribosomes biogenesis and hence is a key target to investigate with respect to biotherapeutic protein synthesis. The amount of recombinant protein produced when these cells are grown in culture is determined by the number of cells that can be generated (the 'biomass') and the amount of recombinant biotherapeutic protein that is produced by each cell (the so-called 'cell specific productivity'). The mammalian target of rapamycin (or mTOR) is a master regulator of both cell growth and proliferation (and hence biomass) and of protein synthesis. However, it remains to be established how this master regulator contributes to recombinant protein synthesis, and whether mTOR can be manipulated to enhance growth and product levels. It is therefore surprising that there has been no comprehensive study of the role of mTOR signalling with respect to the role this might play in determining recombinant protein yields from mammalian cells. We will carry out such a study, investigating our overall hypothesis (i) that the growth and productivity of mammalian recombinant cell lines is underpinned by by mTOR, the global regulator of cell proliferation, ribosome biogenesis and protein synthesis, and (ii) that engineering of this global regulator and signalling pathway increases the rate at which cells divide , and enhances recombinant protein production from CHO cells. The overall aim is to generate new mammalian cells that exploit manipulations of mTOR signalling to enhance the production of monoclonal antibodies and other recombinant products. This information is of very substantial relevance to industry since the production of commercially valuable proteins (e.g. antibodies) is hindered when cells become stressed later in culture and by the amount of biomass accumulated in the bioreactor. Without improved expression systems the biotechnology/pharmaceutical industries will lack the capability to produce large enough amounts of these valuable and effective drugs to meet the demand at a price that will allow them to be prescribed for patients who would benefit from them.
传统上,小分子药物(如抗生素)一直是治疗和治疗男性的主要药物,但在过去10-20年中,基于蛋白质的药物(如赫赛汀,通常用于治疗乳腺癌)已经发展到现在,这些药物构成了药品市场的一个重要部分。有几种基于蛋白质的药物,其中一种,单克隆抗体,在使用或临床试验中构成了一类中数量最多的蛋白质分子。许多基于蛋白质的药物的生产具有挑战性,因为它们(a)需要特定的辅助蛋白质折叠和组装成最终的活性状态,(b)它们的表面被糖和其他对其生物活性至关重要的分子修饰。由于生产这种生物治疗药物需要很高的精度,这种用于治疗疾病的“重组”蛋白质药物通常是由在规定条件下培养的细胞生产的。其中一个问题是,我们用来制造用于治疗的蛋白质的细胞并不像我们希望的那样有效。因此,我们可能无法生产足够的这些药物,或者对卫生保健提供者来说,生产这些药物的成本可能太高。因此,该提案着手解决了一个关键领域,该领域支持从哺乳动物细胞合成重组蛋白。该研究旨在了解蛋白质合成的全球调节因子mTOR(哺乳动物雷帕霉素靶点)如何促进重组蛋白的合成,并设计新的方法来操纵这一过程以提高重组蛋白的产量。蛋白质合成是指细胞内遗传物质DNA中的信息通过中间信使mRNA转化为蛋白质(如抗体)的过程。蛋白质合成参与核糖体,mTOR信号传导也影响核糖体的生物发生,因此是生物治疗蛋白质合成研究的关键目标。当这些细胞在培养基中生长时,产生的重组蛋白的数量取决于可以产生的细胞数量(“生物量”)和每个细胞产生的重组生物治疗蛋白的数量(所谓的“细胞特异性生产力”)。雷帕霉素(或mTOR)的哺乳动物靶标是细胞生长和增殖(因此是生物量)和蛋白质合成的主要调节剂。然而,这个主调控因子如何促进重组蛋白的合成,以及mTOR是否可以被操纵以提高生长和产物水平,仍有待确定。因此,令人惊讶的是,目前还没有对mTOR信号传导在决定哺乳动物细胞重组蛋白产量中的作用进行全面的研究。我们将开展这样一项研究,调查我们的总体假设:(1)哺乳动物重组细胞系的生长和生产力是由mTOR支撑的,mTOR是细胞增殖、核糖体生物发生和蛋白质合成的全局调节剂;(2)这种全局调节剂和信号通路的工程设计提高了细胞分裂的速度,并提高了CHO细胞的重组蛋白产量。总体目标是产生新的哺乳动物细胞,利用mTOR信号的操纵来增强单克隆抗体和其他重组产物的产生。这一信息与工业具有非常重要的相关性,因为当细胞在培养后期受到压力时,以及生物反应器中积累的生物质量会阻碍商业价值蛋白质(例如抗体)的生产。如果没有改进的表达系统,生物技术/制药工业将缺乏生产足够数量的这些有价值和有效的药物的能力,以满足需求,使它们的价格能够开给那些将从中受益的病人。

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)
Cooling-induced SUMOylation of EXOSC10 down-regulates ribosome biogenesis.
冷却诱导的exosc10的Sumoylation下调核糖体生物发生。
  • DOI:
    10.1261/rna.054411.115
  • 发表时间:
    2016-04
  • 期刊:
  • 影响因子:
    0
  • 作者:
    Knight JR;Bastide A;Peretti D;Roobol A;Roobol J;Mallucci GR;Smales CM;Willis AE
  • 通讯作者:
    Willis AE
mTORC1 signalling and eIF4E/4E-BP1 translation initiation factor stoichiometry influence recombinant protein productivity from GS-CHOK1 cells.
  • DOI:
    10.1042/bcj20160845
  • 发表时间:
    2016-12-15
  • 期刊:
  • 影响因子:
    0
  • 作者:
    Jossé L;Xie J;Proud CG;Smales CM
  • 通讯作者:
    Smales CM
Constitutively-active Rheb mutants [T23M] and [E40K] drive increased production and secretion of recombinant protein in Chinese hamster ovary cells
组成型活性 Rheb 突变体 [T23M] 和 [E40K] 驱动中国仓鼠卵巢细胞中重组蛋白的产生和分泌增加
  • DOI:
    10.22541/au.160218204.42443382/v1
  • 发表时间:
    2020
  • 期刊:
  • 影响因子:
    0
  • 作者:
    Poi S
  • 通讯作者:
    Poi S
Control and regulation of mRNA translation.
mRNA 翻译的控制和调节。
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Christopher Smales其他文献

Christopher Smales的其他文献

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{{ truncateString('Christopher Smales', 18)}}的其他基金

Taiwan Partnering Award: Establishing a CHO Cell Expression System for Animal Vaccine Production
台湾合作奖:建立用于动物疫苗生产的CHO细胞表达系统
  • 批准号:
    BB/T01945X/1
  • 财政年份:
    2021
  • 资助金额:
    $ 41.31万
  • 项目类别:
    Research Grant
Generation, characterisation and application of SARS-CoV-2 protein antigens for COVID-19 rapid diagnostic purposes in the hospital and community
SARS-CoV-2 蛋白抗原的生成、表征和应用,用于医院和社区中的 COVID-19 快速诊断
  • 批准号:
    BB/V011324/1
  • 财政年份:
    2020
  • 资助金额:
    $ 41.31万
  • 项目类别:
    Research Grant
An integrated cell and protein engineering approach to generate enhanced CHO cell platforms for manufacture of difficult to express biopharmaceuticals
一种集成的细胞和蛋白质工程方法,用于生成增强的 CHO 细胞平台,用于制造难以表达的生物制药
  • 批准号:
    BB/R001731/1
  • 财政年份:
    2018
  • 资助金额:
    $ 41.31万
  • 项目类别:
    Research Grant
Translation of Step-changing Bioprocesses and Expression System Technologies for Next Generation Protein Biologics Production in CHO Cells
转化用于 CHO 细胞中下一代蛋白质生物制品生产的逐步改变的生物过程和表达系统技术
  • 批准号:
    BB/N023501/1
  • 财政年份:
    2016
  • 资助金额:
    $ 41.31万
  • 项目类别:
    Research Grant
Development and Commercialisation of a Second Generation Rapid Diagnostic Test (RDT) for Human African Trypanosomiasis (HAT) and other Kinetoplastida
针对人类非洲锥虫病 (HAT) 和其他动质体的第二代快速诊断测试 (RDT) 的开发和商业化
  • 批准号:
    BB/N012496/1
  • 财政年份:
    2016
  • 资助金额:
    $ 41.31万
  • 项目类别:
    Research Grant
Feasibility study with the recombinant protein, rISG65, in a new second generation Rapid Diagnostic Test (RDT) for Sleeping Sickness
重组蛋白 rISG65 在新的第二代昏睡病快速诊断测试 (RDT) 中的可行性研究
  • 批准号:
    BB/N004434/1
  • 财政年份:
    2015
  • 资助金额:
    $ 41.31万
  • 项目类别:
    Research Grant
13 ERA IB: Investigating NOvel VAluable bio-Therapeutics and Expression systems
13 ERA IB:研究新颖有价值的生物治疗和表达系统
  • 批准号:
    BB/M000699/1
  • 财政年份:
    2014
  • 资助金额:
    $ 41.31万
  • 项目类别:
    Research Grant
FLIP Expression of recombinant target antigens for neglected tropical diseases in surrogate organisms
FLIP 在替代生物体中表达被忽视的热带病的重组靶抗原
  • 批准号:
    BB/L026279/1
  • 财政年份:
    2014
  • 资助金额:
    $ 41.31万
  • 项目类别:
    Research Grant
Bioprocessing Network: BioProNET
生物处理网络:BioProNET
  • 批准号:
    BB/L013770/1
  • 财政年份:
    2014
  • 资助金额:
    $ 41.31万
  • 项目类别:
    Research Grant
Unravelling and engineering the role of trace metals on recombinant therapeutic protein synthesis and heterogeneity from Chinese hamster ovary cells
揭示和改造微量金属对中国仓鼠卵巢细胞重组治疗性蛋白合成和异质性的作用
  • 批准号:
    BB/K017640/1
  • 财政年份:
    2013
  • 资助金额:
    $ 41.31万
  • 项目类别:
    Research Grant

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冷原子系统自旋压缩的理论研究
  • 批准号:
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Hypothalamic BDNF-mTOR signaling promotes hypertension by increasing cardiovascular sensitivity to stress
下丘脑 BDNF-mTOR 信号通过增加心血管对压力的敏感性促进高血压
  • 批准号:
    10736248
  • 财政年份:
    2023
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    $ 41.31万
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Placental Insulin Signaling and mTOR Nutrient-Sensing Programming of Offspring Metabolic Health
胎盘胰岛素信号传导和 mTOR 营养感应编程对后代代谢健康的影响
  • 批准号:
    10625938
  • 财政年份:
    2022
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mTOR-Mediated Desaturation of Fatty Acids in Hepatic Insulin Resistance.
mTOR 介导的肝胰岛素抵抗中脂肪酸去饱和。
  • 批准号:
    10339318
  • 财政年份:
    2021
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    $ 41.31万
  • 项目类别:
mTOR-Mediated Desaturation of Fatty Acids in Hepatic Insulin Resistance.
mTOR 介导的肝胰岛素抵抗中脂肪酸去饱和。
  • 批准号:
    10554280
  • 财政年份:
    2021
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    $ 41.31万
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mTOR-Mediated Desaturation of Fatty Acids in Hepatic Insulin Resistance.
mTOR 介导的肝胰岛素抵抗中脂肪酸去饱和。
  • 批准号:
    10013714
  • 财政年份:
    2021
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    $ 41.31万
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Mechanisms of membrane protein trafficking and AKT/mTOR signaling that promote myelin sheath stability and growth
促进髓鞘稳定性和生长的膜蛋白运输和 AKT/mTOR 信号传导机制
  • 批准号:
    10328875
  • 财政年份:
    2020
  • 资助金额:
    $ 41.31万
  • 项目类别:
Mechanisms of mTOR-mediated myelin sheath growth
mTOR介导的髓鞘生长机制
  • 批准号:
    9906666
  • 财政年份:
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Determining the mechanism of aspartate sensing by the mTOR pathway
通过 mTOR 途径确定天冬氨酸传感机制
  • 批准号:
    9337243
  • 财政年份:
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  • 资助金额:
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Determining the mechanism of aspartate sensing by the mTOR pathway
通过 mTOR 途径确定天冬氨酸传感机制
  • 批准号:
    9325207
  • 财政年份:
    2017
  • 资助金额:
    $ 41.31万
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Construction of cancer chronotherapy by manipulation of biological clock via mTOR signal pathway.
通过 mTOR 信号通路操纵生物钟构建癌症时间疗法。
  • 批准号:
    15K19167
  • 财政年份:
    2015
  • 资助金额:
    $ 41.31万
  • 项目类别:
    Grant-in-Aid for Young Scientists (B)
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