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

表征 SRCAP 在表皮稳态和鳞状细胞癌中的作用

• 批准号: 10559605
• 负责人: Stephenie Droll
• 金额: $ 4.29万
• 依托单位: NORTHWESTERN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-03-01 至 2025-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10559605
• 关键词: ATPase Domain; Acceleration; Adherence; Adhesions; Binding; Biological Assay; Biological Markers; Biological Sciences; Biology; Biomedical Research; CDK4 gene; Cancer Prognosis; Cancerous; Catalytic Domain; Cell-Cell Adhesion; Cell-Matrix Junction; Cells; ChIP-seq; Chicago; Chromatin; Chromatin Remodeling Factor; Communication; Core Facility; DNA Binding; DNA Damage; DNA Repair; DNA Repair Pathway; Data; Deposition; Development; Disease; ECM receptor; Educational workshop; Epidermis; Epithelium; Equilibrium; Extracellular Matrix; Focal Adhesions; Future; Gene Expression; Gene Expression Regulation; Genes; Genomic approach; Genomics; Histone H2A; Homeostasis; Human; ITGA5 gene; Induced Mutation; Integrins; Intercellular Junctions; Invaded; Kinetics; Knowledge; LacZ Genes; Laminin; Length; Maintenance; Malignant Neoplasms; Mediating; Mentorship; Microscopy; Modeling; Molecular; Mus; Mutate; Mutation; Oncogenes; Operative Surgical Procedures; Pathway interactions; Persons; Proliferating; Proteins; Proteomics; RAS genes; Research; Role; Scaffolding Protein; Skin; Skin Cancer; Skin Carcinoma; Squamous cell carcinoma; Supervision; Testing; Thick; Tissue Model; Tissues; Training; Tumor Weights; United States; Universities; Up-Regulation; Variant; Western Blotting; Xenograft Model; cancer genomics; carcinogenesis; career; cell behavior; cell motility; cofactor; design; epidermal stem cell; experimental study; gene repair; gene repression; genomic data; human tissue; indexing; insight; keratinocyte; knock-down; migration; new therapeutic target; novel; post-doctoral training; programs; protein protein interaction; recruit; skills; skin squamous cell carcinoma; skin xenograft; small hairpin RNA; therapeutic target; transcriptome sequencing; tumor; tumor progression

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.

项目摘要/摘要 皮肤鳞状细胞癌(CSCC)是第二种最常见的恶性肿瘤，约有100万人毁容 每年在美国杀死15,000人。由于CSCC的分子驱动因素不完全 有特点的是，对于手术无法治愈的病例，几乎没有治疗选择。这项研究计划的重点是 在非黑素瘤皮肤癌中高度突变的染色质重构体的特征，但目前 在人体组织中未得到充分研究。第一个目标是用基因组学定义野生型调控机制 原代人类角质形成细胞的研究进展。在上皮性肿瘤中也发现了一个突变热点 现有的癌症基因组数据。基因组和蛋白质组学方法将决定分子机制。 控制这个热点突变是如何导致基因失调的。初步的RNA测序显示， 抑制催化亚单位失调的DNA修复和细胞外基质的组成 (ECM)，与癌症进展有关的通路。因此，目标二将从功能上描述如何 突变会影响癌症的进展。ECM的组成将通过蛋白质印迹进行检测，而 由此引起的表皮粘连的变化将通过免疫染色和显微镜进行探测。ECM通知 细胞行为，如增殖和迁移，这也将被调查。这些结果将揭示 表皮龛和表皮祖细胞的任何癌前改变。最后，一个成熟的， 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 进步。总之，我们的结果将定义一个特性不足的染色质的调节机制 表皮动态平衡和致癌中的重建剂。这些结果将提供功能上的证据 告知SRCAP是否代表一个有用的CSCC生物标记物或一个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，网址为 西北大学由发起人包晓敏博士和联合发起人包小民博士监督。 凯瑟琳·格林，她是表皮生物学专家，拥有支持拟议实验的技能。 培训将利用埃文斯顿和埃文斯顿上可用的研讨会、研讨会和核心设施 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、指导技能和独立性是成功过渡到 博士后培训，最终成为从事生物医学研究的职业。