Determining the Essentiality of Spy-1 as a Novel Regulator of Mammary Cell Development Using CRISPR/Cas9
Standing
Undergraduate
Type of Proposal
Oral Presentation
Faculty
Faculty of Science
Faculty Sponsor
Lisa Porter
Proposal
Breast cancer is the second most common type of cancer in women comprising one in eight new cases of cancer in Canadian women. Mammary development is a tightly regulated process that involves pubertal growth, pregnancy, lactation, and involution. Each of these stages is characterized through unique changes in morphology of mammary glands due to cellular hormonal signaling and regulation of cell cycle progression. The cell cycle is regulated by interactions of cyclins and cyclin-dependent kinases that mediate the passage of the cell through checkpoints that allow for division. Spy1 is an atypical cyclin-like protein that can override cellular damage checkpoints to promote cellular proliferation without requiring cyclin-dependent kinase phosphorylation for activation. Spy1 levels are tightly regulated during mammary gland development with high levels during puberty, early pregnancy, and involution and reduced levels during lactation. This study seeks to understand the essentiality of Spy1 for normal cell proliferation and differentiation during mammary development. Using CRISPR-Cas9, Spy1 was knocked out in the mouse mammary epithelial cell line, HC11. The cell cycle profile for the knockout cell lines was determined to understand the effect of the loss of Spy1 on proliferation. Furthermore, differentiation assays and organoids were used to study the effects of the loss of Spy1 on differentiation. Our results show that the loss of Spy1 results in a delay in the cell cycle as well as increased differentiation. Understanding the role of Spy1 in mammary cell proliferation and differentiation will shed light on the mechanisms regulating normal mammary development, and ultimately allow for better diagnosis and treatment of breast cancer.
Location
Windsor
Grand Challenges
Viable, Healthy and Safe Communities
Determining the Essentiality of Spy-1 as a Novel Regulator of Mammary Cell Development Using CRISPR/Cas9
Windsor
Breast cancer is the second most common type of cancer in women comprising one in eight new cases of cancer in Canadian women. Mammary development is a tightly regulated process that involves pubertal growth, pregnancy, lactation, and involution. Each of these stages is characterized through unique changes in morphology of mammary glands due to cellular hormonal signaling and regulation of cell cycle progression. The cell cycle is regulated by interactions of cyclins and cyclin-dependent kinases that mediate the passage of the cell through checkpoints that allow for division. Spy1 is an atypical cyclin-like protein that can override cellular damage checkpoints to promote cellular proliferation without requiring cyclin-dependent kinase phosphorylation for activation. Spy1 levels are tightly regulated during mammary gland development with high levels during puberty, early pregnancy, and involution and reduced levels during lactation. This study seeks to understand the essentiality of Spy1 for normal cell proliferation and differentiation during mammary development. Using CRISPR-Cas9, Spy1 was knocked out in the mouse mammary epithelial cell line, HC11. The cell cycle profile for the knockout cell lines was determined to understand the effect of the loss of Spy1 on proliferation. Furthermore, differentiation assays and organoids were used to study the effects of the loss of Spy1 on differentiation. Our results show that the loss of Spy1 results in a delay in the cell cycle as well as increased differentiation. Understanding the role of Spy1 in mammary cell proliferation and differentiation will shed light on the mechanisms regulating normal mammary development, and ultimately allow for better diagnosis and treatment of breast cancer.