Project Summary/Abstract Cutaneous squamous cell carcinoma (cSCC) is the second most common malignancy, disfiguring one million and killing 15,000 people per year in the United States. Since the molecular drivers of cSCC are incompletely characterized, few treatment options exist for cases which surgery does not cure. This research plan is focused on characterizing a chromatin remodeler that is highly mutated in non-melanoma skin cancers, but is currently understudied in human tissue. The first aim will define the wild-type regulatory mechanism using genomics approaches in primary human keratinocytes. A mutational hotspot was also identified in epithelial tumors using existing cancer genomics data. Genomic and proteomic approaches will determine the molecular mechanism governing how this hotspot mutation induces gene dysregulation. Preliminary RNA-sequencing revealed that knockdown of the catalytic subunit dysregulated DNA repair and the composition of the extracellular matrix (ECM), pathways implicated in cancer progression. Therefore, aim two will functionally characterize how mutation influences cancer progression. The ECM composition will be examined by western blot while the resulting changes to epidermal adhesion will be probed by immunostaining and microscopy. The ECM informs cellular behaviors such as proliferation and migration, which will also be investigated. These results will reveal any pro-cancerous changes to both the epidermal niche and epidermal progenitors. Finally, an established, inducible cSCC xenograft model will be used to determine whether the hotspot mutation accelerates cancer progression. Altogether our results will define the regulatory mechanism of an under-characterized chromatin remodeler in epidermal homeostasis and carcinogenesis. These results will provide functional evidence informing whether SRCAP represents a useful cSCC biomarker or a worthwhile future therapeutic target. The proposed research will be completed in the Interdisciplinary Biological Sciences graduate program at Northwestern University under the supervision of the sponsor, Dr. Xiaomin Bao, and the co-sponsor, Dr. Kathleen Green, who are experts in epidermal biology with skillsets that support the proposed experiments. Training will take advantage of workshops, seminars, and core facilities available on both the Evanston and Chicago campuses. This research and training plan is designed to develop the applicant's scientific technical, analysis, communication, and mentorship skills and independence necessary to successfully transition to postdoctoral training and ultimately a career conducting biomedical research.