Scaling up the production of cancer organoids for use in drug discovery

类器官在药物开发方面提出了一种潜在的途径;允许减少动物试验，提供更接近人类解剖功能的模型，并提供更复杂的测试机会[1]。在类器官生长中，需要允许3D培养的框架。目前的培养程序使用基质胶形式的支持基质。基质胶由小鼠肉瘤细胞产生。由于高成本和低可用性，因此在工业规模上使用该材料具有挑战性，因为小鼠肉瘤不易在一致的庄园中获得。由于基质胶是一种天然产品，因此它是固有变异性的来源。该项目将专注于开发和测试完全或部分替代合成基质，用于替代或与Matrigel结合使用，为类器官生长提供更一致的环境。所述合成的替代生物材料将需要在类器官生产效率方面至少与单独的基质胶相匹配[2]，[3]。使用合成的替代物在降低批次间差异性的一致性方面带来益处。合成基质可以允许更简单的储存和处理条件，以允许增加的应用和使用。反过来，随着更多条件的可行性，可以增加可调性[4]。随着合成基质的成功使用，可以在成熟过程中为类器官提供增加的结构和机械性能。单独的基质胶不能形成自支撑结构，添加合成基质可以提供这种能力。自支撑结构还将增加类器官生产的可扩展性，提供适合于生物反应器内的生长基础[5]。预期结果是开发一种新型水凝胶，该水凝胶将被测试和优化;导致基质提供工业规模生产的类器官的一致性和质量的增加。在研究中，将研究优化类器官生长的其他途径，包括：测试乳酸盐对类器官生长的影响;以及测试缺氧条件的影响。这些将与合成基质的优化一起，以增加在工业规模上使用类器官的便利性。这些发现将有望最大限度地提高类器官的生长。这项研究的好处在于测试新的潜在药物，即用于治疗不同组织中的癌症。以这种方式进行测试提供了以下方面的机会：医学个性化;更高质量的测试;降低开发成本;减少临床试验中的问题;减少动物测试，这提供了伦理益处，并通过使用人体组织来增加结果的可靠性，这提供了更接近现实的代表性。这些潜在的好处与EPSRC的优先领域相一致，通过医疗保健优化，重点是药物发现，以及旨在提供更高的可负担性，从而实现更具包容性的医疗保健[6]。[1]M. A.兰开斯特和J. A. Knoblich，'Organogenesis in a dish：Modeling development and disease using organoid technologies'，Science，vol. 345，no. 6194，Jul. 2014，doi：10.1126/science.1247125. [2]A. Fatehullah，S. H. Tan和N. Barker，“类器官作为人类发育和疾病的体外模型”，Nat.CellBiol.，第18卷，第3号，第3条，2016年3月，doi：10.1038/ncb 3312。[3]生物材料和组织工程- EPSRC网站。https://epsrc.ukri.org/research/ourportfolio/researchareas/biomaterials/（2020年9月30日访问）。[4]M. C. Kibbey，“EHS肉瘤和基质胶消除的维持”，J. Tissue Cult。Methods，vol. 16，no. 3，pp. 227-230，1994年9月，doi：10.1007/BF 01540656。[5]N. Gjorevski等人，“用于肠干细胞和类器官培养的设计基质”，Nature，第539卷，第7630号，第7630号，2016年11月，doi：10.1038/nature 20168。[6]K.丹尼尔，“EPSRC医疗保健技术战略摘要”，第12页

Organoids pose a potential route in terms of drug development; allowing a reduction in animal testing, provision of a model more closely resembling human anatomical function, and presenting more complex testing opportunities[1].In organoid growth a framework to allow 3D culture is required. Current culture procedure uses a supporting matrix in the form of Matrigel. Matrigel is produced from mouse sarcoma cells. The use of this at an industrial scale is challenging due to high cost, and low availability as mouse sarcoma is not easily sourced in a consistent manor.As Matrigel is a natural product it is a source of inherent variability. This project will focus on the development and testing of a full or partial replacement synthetic matrix to be used in place of or in combination with Matrigel to provide a more consistent environment for organoid growth. Said synthetic alternative biomaterial will be required to at least match Matrigel alone in efficiency of organoid production[2], [3].Use of a synthetic alternative brings benefits in terms of consistency with reduced batch to batch variability. A synthetic matrix could allow more simplistic storage and handling conditions to allow increased application, and usage. In turn could increase tunability as more conditions would be viable[4].With the successful use of synthetic matrix, increased structural and mechanical properties could be provided to the organoids during maturation. Matrigel alone cannot form a self-supporting structure, the addition of a synthetic matrix could provide this ability. A self-supporting structure would also increase the scalability of organoid production providing a growth foundation which would be suitable within a bioreactor[5].The intended result is development of a novel hydrogel which will be tested and optimised; resulting in a matrix providing an increase in both consistency, and quality of organoids produced on an industrial scale. Within the research other avenues of optimisation of organoid growth will be investigated including: testing the effect of lactate on organoid growth; and testing the effect of hypoxic conditions. These will sit alongside the optimisation of a synthetic matrix to increase the ease of organoid use on an industrial scale.These finding will hope to maximise organoid growth. Benefits of this research lie in testing of new potential drugs namely for the treatment of cancers within differing tissues. Testing this way provides opportunities in personalisation of medicine; higher quality testing; reduction in development costs; decreased issues in clinical trials; and reduction in animal testing which provides ethical benefits as well as increasing the reliability of results through the use of human tissue which provides closer representation to actuality. These potential benefits align with the EPSRC priority areas through healthcare optimisation focusing on drug discovery, alongside the intention to provide increased affordability resulting in more inclusive healthcare[6].[1]M. A. Lancaster and J. A. Knoblich, 'Organogenesis in a dish: Modeling development and disease using organoid technologies', Science, vol. 345, no. 6194, Jul. 2014, doi: 10.1126/science.1247125.[2]A. Fatehullah, S. H. Tan, and N. Barker, 'Organoids as an in vitro model of human development and disease', Nat. Cell Biol., vol. 18, no. 3, Art. no. 3, Mar. 2016, doi: 10.1038/ncb3312.[3]'Biomaterials and tissue engineering - EPSRC website'. https://epsrc.ukri.org/research/ourportfolio/researchareas/biomaterials/ (accessed Sep. 30, 2020).[4]M. C. Kibbey, 'Maintenance of the EHS sarcoma and Matrigel preparation', J. Tissue Cult. Methods, vol. 16, no. 3, pp. 227-230, Sep. 1994, doi: 10.1007/BF01540656.[5]N. Gjorevski et al., 'Designer matrices for intestinal stem cell and organoid culture', Nature, vol. 539, no. 7630, Art. no. 7630, Nov. 2016, doi: 10.1038/nature20168.[6]K. Daniel, 'EPSRC Healthcare Technologies Strategy Summary', p.12