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.

PHYSICAL ACTIVITY EXPERIENCES OF PERSIAN WOMEN RECENTLY IMMIGRATED TO CANADA

New immigrants to Canada are generally in similar or better physical and mental health than people born in Canada, however, many studies suggest that their health tends to decline quickly after immigration. Lower physical activity levels among new immigrants might be contributing to this phenomenon. There is a paucity of information regarding the physical activity behaviour of the Canadian immigrant population in general and of West Asian women, such as Iranians (Persians), in particular. Given that this group is characterised by an increasing population growth and lower rates of physical activity, it is critical to understand how best to address physical activity promotion in this population. Purpose: To understand the physical activity experiences of Persian women recently immigrated to Toronto, Canada in order to develop recommendations for the design and implementation of tailored physical activity programs. Methods: A qualitative interpretive description approach was chosen to collect and describe ideas, experiences, and perceptions of physical activity within 10 new immigrant women. Using an inductive approach, two fundamental techniques of immersion and crystallization were used throughout the analysis. Thematic analysis was conducted by performing a sequential process of open and axial coding. Emerged themes were further conceptualized through a socio-ecological lens. Results: The facilitators and barriers to physical activity among the women were situated within five overarching categories, 1) Perceptions about physical activity, 2) New physical environment and social structure, 3) Cultural heritage values, 4) Settlement and immigration factors, and 5) Physical activity program features. Discussion: Findings revealed that Persian new immigrant women’s engagement in physical activity after immigration is influenced by factors across the individual, sociocultural, environmental, institutional, and policy levels. Newcomer women’s physical activity was influenced by their transition from their society of origin to the host society and the challenges and successes experienced throughout the settlement and acculturation process. The most noticeable barrier to physical activity in Canada for the women emerged as the lack of communication of physical activity information to newcomers. A set of recommendations is provided for developing efficient physical activity programs for Persian immigrant women, which may also be relevant for other immigrant groups in Canada.