Towards the manipulation of the UPR in mammalian cells to orchestrate cellular re-organization for enhanced monoclonal antibody production

我们已经开发出了一种防御机制，当我们的身体识别出外国“入侵者”时，就会对感染做出反应。我们体内有一种被称为B细胞的细胞，这种细胞在受到感染时会变成产生抗体的细胞。这是必要的，使我们能够抵御感染，否则将是非常有害的。抗体通过攻击并摧毁外来入侵者来发挥作用，从而清除感染并清除外来病原体。为了让B细胞转变为抗体产生细胞(这一过程被称为分化)，它们必须调整自身的组织结构，以便拥有制造大量抗体所需的细胞器。由于抗体是人体对疾病的天然防御，许多新的抗体类型的药物正在开发中，以帮助治疗人类疾病，如癌症和艾滋病。然而，为了生产这些基于抗体的下一代治疗性药物，我们必须使用哺乳动物细胞来制造它们。我们用来制造这些药物的细胞类型在产生抗体方面不如改良的B细胞有效，因此我们无法生产足够的这些药物，因此对它们的成本和需求很高。人们认为，随着更多基于抗体的药物的开发，这将成为一个更大的问题。这里提出的研究将检验我们是否可以找到具有更多所需机械来产生高水平抗体的哺乳动物细胞，或者，我们是否可以操纵这些哺乳动物细胞来产生更多这种机械，以便以更低的成本更快地生产出更高产量或数量的这些药物。目前还不清楚这是否可能，而且用来产生这些抗体的哺乳动物细胞对这一过程也知之甚少。被称为蛋白质组学和诱导表达技术的先进技术将被用来研究已知对分化的B细胞产生抗体至关重要的蛋白质水平的差异，并比较用于商业抗体生产的哺乳动物细胞中这些蛋白质的水平。我们将寻找变得更丰富或更少的蛋白质(通过改变基因表达、蛋白质合成和/或蛋白质降解)，并对已知能够改变其功能的先前存在的蛋白质进行微妙的分子修饰(例如，打开或关闭它们)。来自当前基因组学项目的信息将被挖掘出来，并与我们的蛋白质数据结合使用，以确定如何提高我们可以使用这些哺乳动物细胞制造的治疗性蛋白质“药物”的数量。如上所述，这是极其重要的，因为预计随着越来越多的蛋白质药物的开发，我们将缺乏生产足够大的数量来满足对这些新药的需求的能力。

We have developed a defence mechanism to respond to infection when our body recognises a foreign 'invader'. We have a type of cell known as a B cell in our body, which in response to infection changes into an antibody-producing cell. This is necessary to allow us to fight off infections that would otherwise be very harmful. The antibodies work by attacking the foreign invader and destroying it, thereby clearing the infection and removing the foreign agent. In order for the B cells to change into antibody producing cells (a process known as differentiation), they must adjust the organization of themselves so that they have the required pieces of cellular machinery to make large amounts of antibody. As antibodies are the bodies natural defence against disease, many new antibody type drugs are being developed to help treat human diseases such as cancer and AIDS. However, in order to produce these next generation antibody-based therapeutic 'drugs' we must use mammalian cells to make them. The types of cells we use to make these drugs are not as efficient at producing antibodies as the modified B cell and as a result we are not able to produce enough of these drugs and the cost and demand for them is therefore high. It is thought that this will become even more of a problem as more antibody based drugs are developed. The research proposed here will examine whether we can find mammalian cells with more of the required machinery to produce high-levels of antibodies, or alternatively, if we can manipulate these mammalian cells to produce more of this machinery so that higher yields or amounts of these drugs can be produced more quickly at less cost. At present it is unknown if this is possible, and the process is poorly understood in the mammalian cells used to produce these antibodies. Advanced technology known as proteomics and inducible expression technology will be used to study the differences in the levels of the proteins known to be important for antibody production in differentiated B cells and compare the levels of these proteins in the mammalian cells used for commercial antibody production. We will look for proteins that become either more or less abundant (by altered gene expression, protein synthesis and/or protein degradation) and for subtle molecular modifications to pre-existing proteins known to be able to modify their function (e.g., switch them on or off). Information from current genomics projects will be mined and used in combination with our protein data to identify ways of improving the amount of therapeutic protein 'drug' we can manufacture using these mammalian cells. As stated above, this is extremely important as it is expected that with an increasing number of protein 'drugs' being developed we will lack the capability of producing large enough amounts to meet the required demand for these new drugs.