Characterization of PAD4 Inhibition and its Anti-Cancer Effects via mTOR Pathway
Open Access
Author:
Xu, Haonan
Area of Honors:
Biochemistry and Molecular Biology
Degree:
Bachelor of Science
Document Type:
Thesis
Thesis Supervisors:
Yanming Wang, Thesis Supervisor Joseph C. Reese, Thesis Honors Advisor
Keywords:
PAD4 Autophagy Cancer CRISPR
Abstract:
PAD4 belongs to a family of enzymes that converts arginine and monomethyl arginine into citrulline. Citrullination of histone molecules can cause various transcriptional modulations, and the overexpression of PAD4 in neutrophil has been linked to many diseases. In the case of cancer, it was shown that inhibition of PAD4 can induce ER stress, trigger autophagy, and cause cancer cell death. In this study, two in-house PAD4 inhibitors GSK484-07 and YW4-03 have demonstrated potent effects on killing triple negative breast cancer cells. The treatments led to an upregulation of SESN2 signal, which suggests possible relation to mTOR mediated autophagy. Furthermore, YW4-03 was shown to increase HO-1 expression, which is indicative of oxidative stress. To further understand the anticancer mechanisms in which PAD4 is involved in, a CRISPR/Cas9 based human PAD4 knockout construct was created, despite the limited success in generating single colony knockout cell lines.
