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Drug Discovery


these advances are derived from the basic under- pinning science, which originated three decades ago. Researchers have devised new methods to utilise different types of stem cells to generate organoids resembling a multitude of different organs. In 2009, the laboratory of Hans Clevers at the Hubrecht Institute demonstrated that single LGR5-responsive stem cells can build crypt-villus structures in vitro, opening up the field to produce immortal colorectal organoid lines6. This emerging science is driving the growing interest in organoid- based drug discovery, especially in colorectal can- cer. Some examples of the different morphologies observed in such colorectal cancer organoid lines are shown in Figure 2, which were grown from patient-derived tumours. Currently, specialist academic research labora-


Figure 4 Organoids have been treated


with a range of concentrations of trametinib as described in


the main text and then fixed in situ and the Promega CellTiter-Glo® 3D Cell Viability Assay was used to determine cell viability in this


colorectal cancer organoid cell line. The assay measured ATP


as an indicator of viability and generated a luminescent readout


tories use organoids that are grown manually for basic research, but production is both costly and labour-intensive. However, organoids are not typ- ically produced in the quantities required for the widespread use in drug discovery and lack the uni- formity of size and metabolic standardisation required to generate the high-quality data required to drive effective decision making in the drug dis- covery process. There is a need to grow cancer organoids on an industrial scale for incorporation into both medium- and high-throughput screening protocols and the key features and benefits of this approach are illustrated in Figure 3. To solve the problem created by this bottleneck, organoid expansion processes that can grow organoids in the medium- to high-volume range required for the pharmacological profiling of hundreds to thousands of compounds have been developed7. Incorporating these new technologies into a larg- er-scale bioprocess has developed a capability to considerably increase the productivity of individu- al scientists working manually. Therefore, with the increased availability of bioprocess-expanded organoids being provided at scale in cancer thera- peutics research, the use of tumour-derived


DMSO control Low


Trametinib concentration Mid


organoids as novel research tools is starting to dis- rupt the early-stage drug discovery process. By potentially improving the predictive power of early compound screening, this new approach could provide a solution for the unsustainably high rate of compound attrition in both drug dis- covery and development, especially for novel can- cer therapeutics. The provision of expanded organoids as


research tools can now be reliably supplied as frozen organoid lines in cryovials, ready for seed- ing into various assay formats for drug discovery. The usual requirement is for compound screening being performed in 96- or 384-well screening for- mats in conventional microtitre plates. Typical data obtained from an example of a patient- derived colorectal organoid line are shown in Figure 4, where pharmacological characterisation with a known colorectal cancer drug, trametinib, demonstrates the expected response to this kinase inhibitor. The thawed organoids were resuspended in a hydrogel in 96-well plates and incubated at 37˚C and exposed to trametinib for two days before being assessed for cell viability, as described in the figure legend. Consequently, with bioprocessing technology


required to expand human cancer organoids now available, patient-derived organoids can be used to meet the emerging demand for the large-scale pro- duction of organoids in cancer drug discovery. These organoids can then be used in assay formats compatible with high-throughput screening.DDW


Dr Mark Treherne has been actively involved in the biopharmaceutical industry for more than 25 years and previously led the neurodegeneration research group at Pfizer’s research facility in Sandwich, including using stem-cell derived lines for screening compounds. In 1997, he co-founded Cambridge Drug Discovery as Chief Executive, leading the company’s subsequent acquisition by BioFocus plc,


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Drug Discovery World Winter 2018/19


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