Kinome-wide CRISPR-Cas9 screen to identify kinases involved in Taxol resistance in breast cancer

Breast cancer is the most frequently diagnosed cancer in women, accounting for over 25% of cancer diagnoses and 13% of cancer-related deaths in Canadian women. There are many types of therapies for treatment or management of breast cancer, with chemotherapy being one of the most widely used. Taxol (paclitaxel) is one of the most extensively used chemotherapeutic agents for treating cancers of the breast and numerous other sites. Taxol stabilizes microtubules during mitosis, causing the cell cycle to arrest until eventually the cell undergoes apoptosis. Although Taxol has had significant benefits in many patients, response rates range from only 25-69%, and over half of Taxol-treated patients eventually acquire resistance to the drug. Drug resistance remains one of the greatest barriers to effective cancer treatment, yet little has been discerned regarding resistance to Taxol, despite its widespread clinical use. Kinases are known to be heavily involved in cancer development and progression, and several kinases have been linked to resistance of Taxol and other chemotherapeutic agents. However, a systematic screen for kinases regulating Taxol resistance is lacking. Thus, in this study, a set of kinome-wide screens was conducted to interrogate the involvement of kinases in the Taxol response. Positive-selection and negative-selection CRISPR-Cas9 screens were conducted, whereby a pooled library of 5070 sgRNAs targeted 507 kinase-encoding genes in MCF-7 breast cancer cells that were Taxol-sensitive (WT) or Taxol-resistant (TxR) which were then treated with Taxol. Next generation sequencing (NGS) was performed on cells that survived Taxol treatment, allowing identification and quantitation of sgRNAs. STK38, Blk, FASTK and Nek3 stand out as potentially critical kinases for Taxol-induced apoptosis to occur. Furthermore, kinases CDKL1 and FRK may have a role in Taxol resistance. Further validation of these candidate kinases will provide novel pre-clinical data about potential predictive biomarkers or therapeutic targets for breast cancer patients in the future.

Performance Information and Innovation in the Canadian Government (Working Paper 43)

The presentation made at the conference addressed the issue of linkages between performance information and innovation within the Canadian federal government1. This is a three‐part paper prepared as background to that presentation. • Part I provides an overview of three main sources of performance information - results-based systems, program evaluation, and centrally driven review exercises – and reviews the Canadian experience with them. • Part II identifies and discusses a number of innovation issues that are common to the literature reviewed for this paper. • Part III examines actual and potential linkages between innovation and performance information. This section suggests that innovation in the Canadian federal government tends to cluster into two groups: smaller initiatives driven by staff or middle management; and much larger projects involving major programs, whole departments or whole-of-government. Readily available data on smaller innovation projects is skimpy but suggests that performance information does not play a major role in stimulating these initiatives. In contrast, two of the examples of large-scale innovation show that performance information plays a critical role at all stages. The paper concludes by supporting the contention of others writing on this topic: that more research is needed on innovation, particularly on its link to performance information. In that context, other conclusions drawn in this paper are tentative but suggest that the quality of performance information is as important for innovation as it is for performance management. However, innovation is likely to require its own particular performance information that may not be generated on a routine basis for purposes of performance management, particularly in the early stages of innovation. And, while the availability of performance information can be an important success factor in innovation, it does not stand alone. The commonality of a number of other factors identified in the literature surveyed for this paper strongly suggests that equal if not greater priority needs to be given to attenuating factors that inhibit innovation and to nurturing incentives.