Characterizing the role of SRCAP in Epidermal Homeostasis and Squamous Cell Carcinoma

项目总结/摘要 皮肤鳞状细胞癌是第二常见的恶性肿瘤， 每年在美国杀死15,000人。由于cSCC的分子驱动因素不完全 由于其特征，对于手术不能治愈的病例存在很少的治疗选择。这项研究计划的重点是 在非黑色素瘤皮肤癌中高度突变的染色质重塑，但目前 在人体组织中研究不足。第一个目标将使用基因组学来定义野生型调控机制 在原代人角质形成细胞中的方法。在上皮性肿瘤中也发现了一个突变热点， 现有的癌症基因组学数据。基因组学和蛋白质组学方法将确定分子机制 控制热点突变如何导致基因失调。初步的RNA测序显示， 催化亚基的敲低失调的DNA修复和细胞外基质的组成 (ECM)，与癌症进展有关的途径。因此，目标二将从功能上描述如何 突变影响癌症进展。ECM组合物将通过蛋白质印迹法检查，而ECM组合物将通过免疫印迹法检查。 将通过免疫染色和显微镜检查来探测由此产生的表皮粘附的变化。ECM通知 细胞行为，如增殖和迁移，这也将被研究。这些结果将揭示 表皮小生境和表皮祖细胞的任何癌前变化。最后，一个既定的， 可诱导的cSCC异种移植模型将用于确定热点突变是否加速癌症 进展总之，我们的研究结果将确定一个不足的特点染色质的调控机制 在表皮稳态和癌发生中的重塑。这些结果将提供功能性证据 告知SRCAP是否代表有用的cSCC生物标志物或有价值的未来治疗靶点。 拟议的研究将在跨学科生物科学研究生课程中完成， 西北大学的监督下，申办者，博士Xiaomin鲍，和共同申办者，博士。 凯瑟琳绿色，谁是表皮生物学专家与技能，支持拟议的实验。 培训将利用埃文斯顿和 芝加哥校园。这项研究和培训计划旨在发展申请人的科学技术， 分析，沟通和指导技能和独立性，成功过渡到 博士后培训，并最终从事生物医学研究。

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.