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: 10363661
• 负责人: Stephenie Droll
• 金额: $ 4.2万
• 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/10363661
• 关键词: ATPase Domain; Adherence; Adhesions; Back; 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; Down-Regulation; ECM receptor; Educational workshop; Epidermis; Epithelial; 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; Kinetics; Knowledge; LacZ Genes; Laminin; Length; Maintenance; Malignant Neoplasms; Mediating; Mentorship; Microscopy; Modeling; Molecular; Mus; Mutate; Mutation; Oncogenes; Operative Surgical Procedures; Pathway interactions; Persons; Proteins; Proteomics; RAS genes; Research; Research Training; 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; chromatin remodeling; cofactor; design; epidermal stem cell; experimental study; gene repair; 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.

项目概要/摘要 皮肤鳞状细胞癌 (cSCC) 是第二常见的恶性肿瘤，导致一百万人毁容 每年在美国造成 15,000 人死亡。由于cSCC的分子驱动因素不完全 其特点是，对于手术无法治愈的病例，几乎没有治疗选择。本研究计划重点 表征在非黑色素瘤皮肤癌中高度突变的染色质重塑蛋白，但目前 在人体组织中进行了深入研究。第一个目标是利用基因组学定义野生型调控机制 原代人角质形成细胞中的方法。使用以下方法在上皮性肿瘤中还发现了突变热点 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 控制这种热点突变如何诱导基因失调。初步 RNA 测序表明 抑制催化亚基失调的 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，目标二将在功能上描述如何 突变影响癌症进展。 ECM 成分将通过蛋白质印迹进行检查，同时 将通过免疫染色和显微镜来探测由此产生的表皮粘附变化。 ECM 通知 增殖和迁移等细胞行为也将被研究。这些结果将揭示 表皮生态位和表皮祖细胞的任何促癌变化。最后，一个既定的、 诱导性cSCC异种移植模型将用于确定热点突变是否加速癌症 进展。总而言之，我们的结果将定义未充分表征的染色质的调控机制 表皮稳态和致癌作用的重塑者。这些结果将提供功能证据 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，在发起人鲍晓敏博士和共同发起人鲍晓民博士的监督下。 凯瑟琳·格林（Kathleen Green）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提供的讲习班、研讨会和核心设施 芝加哥校区。该研究和培训计划旨在培养申请人的科学技术、 成功转型所需的分析、沟通、指导技能和独立性 博士后培训并最终从事生物医学研究。