Investaging the role of KIFC1 and centrosome amplification in metastatic uveal melanoma : Towards building a protac to target KIFC1

Patel, Umar and Gadd, Morgan and Fielding, Andrew (2019) Investaging the role of KIFC1 and centrosome amplification in metastatic uveal melanoma : Towards building a protac to target KIFC1. Masters thesis, Lancaster University.

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Abstract

Uveal melanoma is melanoma of the uveal tract, which develops into liver metastasis in approximately 50% of patients. Lack of therapeutic targets means that survival from metastatic uveal melanoma remains at 8% after two years, a survival rate that had not improved since 1973 (Singh et al., 2011). Recent in silico genomics research has revealed uveal melanoma to express genes associated with centrosome amplification (de Almeida et al., 2019). Although centrosome amplification causes cancer (Levine et al., 2017), it has been described as an “Achilles’ heel” for cancer cells as they require mechanisms to cluster their centrosomes to avoid multipolar mitosis, aneuploidy and cell death (Kwon et al., 2008). KIFC1, a kinesin-14 motor protein, is essential for clustering centrosomes and avoiding multipolar mitosis (Kwon et al., 2008). Frequently found to be overexpressed in cancers with centrosome amplification, KIFC1 overexpression has been shown to increase proliferation, survival and invasiveness (Pannu et al., 2015b), but seems to be dispensable in normal somatic cells (Watts et al., 2013). Small molecule KIFC1 inhibitors do exist, but current options display off target effects and require high doses to generate an effect (Yukawa et al., 2018). In this project, metastatic uveal melanoma cell lines have been shown to display centrosome amplification for the first time. Furthermore, KIFC1 depletion in primary and uveal melanoma cell lines reduces proliferation, clonogenicity and survival while increasing the frequency of apoptosis and multipolar mitosis. The addiction to KIFC1 observed in primary uveal melanoma cells, which have insignificant levels of centrosome amplification, implies that KIFC1 has centrosome clustering-independent oncogenic activity in primary uveal melanoma and may be a novel therapeutic target. A novel therapeutic alternative to small molecule inhibitors is the use of small-molecule PROTACs, which are bifunctional chimeras that hijack the ubiquitin-proteasome system to induce degradation of disease-causing target proteins, while also working at lower concentrations and with better specificity than the small molecule inhibitors that they are derived from (Buhimschi et al., 2018). In this project, analogues of the KIFC1 inhibitor CW069 were synthesised as the first step towards producing KIFC1-targetting PROTACs.