Transcriptional kinases in cancer and developmental disorders

Cyclin-dependent kinases (CDKs) are protein kinases performing essential functions in regulation of cell cycle and transcription. Each CDK requires an interaction with a specific cyclin partner for switching on their kinase activity. Mutations in different CDKs or associated cyclins have been linked to developmental defects. For example, mutations in CDK10 and in cyclin Q are both causing developmental disorders, named Al Kaissi syndrome and STAR syndrome, respectively, that are only partially overlapping. In addition, CDK10 is misregulated and mutated in several types of cancer, including breast and gastro-intestinal cancer. For example, CDK10 has been linked to tamoxifen resistance in breast cancer through its regulation of the ETS2 protein; a transcription factor which can promote cellular proliferation, invasion, and metastasis in cancers. However, the functions of CDK10 and its associated cyclin Q (previously cyclin M) are poorly understood, mostly due to the absence of specific inhibitors.

To determine the functions of CDK10, we used CRISPR/Cas9 genome engineering to generate a human cancer cell line in which the endogenous CDK10 can be selectively and rapidly inhibited using a bulky ATP analogue. Preliminary data has shown that CDK10 inhibition affects gene expression in a potentially ETS2-independent manner, indicating a potential direct role of CDK10 on transcription and/or RNA processing. The CDK10 analogue-sensitive cell line will be therefore used to define by phospho-proteomics the proteins phosphorylated by CDK10 and the effects of CDK10 inhibition on different cellular processes, including cell cycle and gene expression.

CRISPR/Cas9 will also be used to generate novel cell lines with the Al Kaissi and STAR pathogenic mutations found in CDK10 and cyclin Q to understand the cellular consequences of these mutations. This project will provide important insights into the mechanisms behind these two developmental disorder syndromes and will determine if the cellular consequences of CDK10 and cyclin Q mutations are due only to a loss of CDK10 activity or also from additional reasons.

Investigating the roles of CDK10 in ETS2 factor stability and phosphorylation of other targets provides the opportunity to exploit our expertise in basic research to identify druggable pathways regulated by CDK10 for potential clinical benefit. This project will also determine whether inhibiting CDK10 could be clinically relevant for the treatment of specific cancers (gastro-intestinal, cutaneous melanomas, and breast) and pave the way for the development of CDK10 specific inhibitors.

Techniques for this project:

Human cell culture, CRISPR/Cas9, phospho-proteomics, RNA-seq, nascent transcription techniques (POINT-seq, PRO-seq), flow cytometry, immunofluorescence, bioinformatics, and standard molecular biology techniques (western blot, co-immunoprecipitation, cloning, PCR, qPCR, …). 

Entry requirements

• Those who have a 1st or a 2.1 undergraduate degree in a relevant field are eligible.
• Evidence of quantitative training is required. For example, AS or A level Maths, IB Standard or Higher Maths, or university level maths/statistics course.
• Those who have a 2.2 and an additional Masters degree in a relevant field may be eligible.
• Those who have a 2.2 and at least three years post-graduate experience in a relevant field may be eligible.
• Those with degrees abroad (perhaps as well as postgraduate experience) may be eligible if their qualifications are deemed equivalent to any of the above.
• University English language requirements apply.

To apply

Carefully read the application advice on our website below and submit your PhD application. 

https://le.ac.uk/study/research-degrees/research-subjects/molecular-and-cell-biology