Regulators of Epithelial Tumor Progression

上皮肿瘤进展的调节因子

• 批准号: 9033595
• 负责人: PAUL KHAVARI
• 金额: --
• 依托单位: VETERANS ADMIN PALO ALTO HEALTH CARE SYS
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-04-01 至 2020-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9033595
• 关键词: Affect; Aneuploidy; Architecture; Cell Cycle Progression; Cell division; Cells; Cessation of life; Characteristics; Chromatids; Clinical; Collagen; Collagen Gene; Collagen Type XI; Complex; Country; Cutaneous; DNA Sequence Alteration; Development; Disease; Dominant-Negative Mutation; Environment; Epidermolysis Bullosa Dystrophica; Epithelial; Epithelial-Stromal Communication; Epithelium; Extracellular Matrix; Extracellular Matrix Proteins; Extracellular Protein; Foundations; Frequencies; Funding; Future; Gene Frequency; Generations; Genes; Genomic Instability; Glycine; Human; Indium; Individual; Inherited; Intervention; Kinetochores; Label; Laboratories; Lead; Light; Malignant Neoplasms; Mass Spectrum Analysis; Military Personnel; Minority; Modeling; Morbidity - disease rate; Mutate; Mutation; Nature; Neoplasms; Oncogenic; Pathogenesis; Patients; Physiological; Plague; Population; Process; Proline; Protein Family; Proteins; Recurrence; Role; Services; Set protein; Skin; Specimen; Squamous cell carcinoma; Sun Exposure; Sunlight; Testing; Tissue Model; Tissues; Translating; UV Radiation Exposure; Veterans; anticancer research; base; cancer prevention; cancer therapy; carcinogenesis; cell stroma; cohesion; deep sequencing; design; exome sequencing; extracellular; genome integrity; human tissue; in vivo; insight; melanoma; mutant; neoplastic; neoplastic cell; novel; public health relevance; skin squamous cell carcinoma; sunlight-induced; tumor; tumor progression; tumorigenesis; ultraviolet

 DESCRIPTION (provided by applicant): The development of epithelial cancers in multiple tissues, including the skin, is characterized by abnormalities that are both intrinsic to the cell as well as outside the cell in the extracellular environment. During the most recent funding cycle of this Merit Review, we characterized the mutational spectrum of cutaneous squamous cell carcinoma (SCC), the second most common cancer in the country, long- associated with sunlight exposure and widespread impacts on the U.S. veteran population. Deep sequencing of 100 SCCs and 100 patient-matched normal skin specimens identified highly recurrent mutations in the COL11A1 gene (52% of SCCs), which encodes a secreted collagen chain involved in epithelial-stromal interactions as well as novel ultraviolet (UV)-signature mutations in the intracellular, kinetochore gene, KNSTRN (19% of SCCs). A major role for mutant extracellular collagen chains and mutant kinetochore proteins has not been widely appreciated in cancer before and their presence in SCC may shed light on fundamental cancer processes. This competing renewal will therefore characterize the role of these genes in epidermal tumor progression. First, we will determine the function of recurrently mutated collagen genes in epidermal tumorigenesis. We found collagen chain mutations, concentrated at glycine or proline substitutions in the Gly-X-Y motif (with X and Y commonly proline) alpha helical collagen sequence, in 70% of SCCs, with mutation of COL11A1 the most common. In inherited monogenic human disorders, such as dystrophic epidermolysis bullosa (DEB), comparable mutations in COL7A1 produce dominant-negative collagens that disrupt epithelial-stromal cohesion, suggesting a model in which tumors secrete mutant collagens to induce neoplasia-enabling abnormalities in the surrounding stroma. To explore this possibility, we will generate human epidermal tissue with defined mutant collagens in vivo then examine the resulting functional impact on epidermal invasion and tumorigenesis. Aim I will thus test a model in which secreted mutant collagens disrupt native extracellular matrix to facilitate neoplastic progression. Second, we will define the identity and function of Kinastrin-interacting proteins as well as characterize recurrently mutated kinetochore genes in epidermal tumor progression. We observed that the KinastrinS24F UV-signature hotspot mutant prevalent in SCC accelerates epidermal tumorigenesis in vivo. In doing so, it exerts impacts characteristic of cancer, specifically increased cell division and genomic instability. These impacts are distinct and separable, however, suggesting separate mechanisms of action for each effect. To explore this, we will identify the protein interactome of both wild-type and UV-hotspot mutant KinastrinS24F proteins by vicinal protein labeling and mass spectrometry. Kinastrin-interacting proteins will then be functionally disrupted and the resulting impacts on KinastrinS24F-enhanced tumorigenesis will be determined. Also, given that 77% of SCCs were found to have mutations in additional core kinetochore genes, we also plan to disrupt selected kinetochore genes and study their impact on epidermal tumorigenesis. Aim II will thus test a model in which mutant Kinastrin interacts with discrete sets of proteins to enhance tumor progression and in which mutations in specific other kinetochore genes enable carcinogenesis. At the end of proposed funding, we plan to have gained further insight into mechanisms of epidermal tumor progression as a foundation for future strategies for cancer prevention and treatment.

 描述（由申请人提供）： 包括皮肤在内的多种组织中上皮癌的发展特征是细胞固有的以及细胞外环境中的异常。在本次优异评审的最新资助周期中，我们描述了皮肤鳞状细胞癌（SCC）的突变谱，这是美国第二常见的癌症，与阳光照射长期相关并对美国退伍军人群体产生广泛影响。对 100 个鳞状细胞癌和 100 个患者匹配的正常皮肤标本进行深度测序，发现了 COL11A1 基因（占鳞状细胞癌的 52%）中高度复发的突变，该基因编码参与上皮-基质相互作用的分泌胶原链，以及细胞内动粒基因 KNSTRN（占鳞状细胞癌的 19%）中的新型紫外线 (UV) 特征突变。突变细胞外胶原链和突变动粒蛋白在癌症中的主要作用此前尚未得到广泛认识，它们在鳞状细胞癌中的存在可能有助于揭示基本的癌症过程。因此，这种竞争性更新将表征这些基因在表皮肿瘤进展中的作用。 首先，我们将确定反复突变的胶原蛋白基因在表皮肿瘤发生中的功能。我们在 70% 的鳞状细胞癌中发现了胶原蛋白链突变，集中在 Gly-X-Y 基序（X 和 Y 通常为脯氨酸）α 螺旋胶原序列中的甘氨酸或脯氨酸取代，其中 COL11A1 突变最常见。在遗传性单基因人类疾病中，例如营养不良性大疱性表皮松解症 (DEB)，COL7A1 中的类似突变会产生显性失活的胶原蛋白，破坏上皮基质的凝聚力，这表明肿瘤分泌突变胶原蛋白以诱导周围基质中发生肿瘤的异常。为了探索这种可能性，我们将在体内产生具有确定的突变胶原的人类表皮组织，然后检查由此产生的对表皮侵袭和肿瘤发生的功能影响。因此，我将测试一个模型，其中分泌的突变胶原蛋白破坏天然细胞外基质以促进肿瘤进展。 其次，我们将定义动素相互作用蛋白的身份和功能，以及表征表皮肿瘤进展中反复突变的动粒基因。我们观察到 SCC 中普遍存在的 KinastrinS24F 紫外线特征热点突变体加速了体内表皮肿瘤的发生。在此过程中，它发挥了癌症特征的影响，特别是增加了细胞分裂和基因组不稳定性。然而，这些影响是不同且可分离的，表明每种影响有不同的作用机制。为了探索这一点，我们将通过邻近蛋白标记和质谱法鉴定野生型和紫外线热点突变体 KinastrinS24F 蛋白的蛋白质相互作用组。然后，Kinatrin 相互作用蛋白将被功能性破坏，并确定由此产生的对 KinastrinS24F 增强的肿瘤发生的影响。此外，鉴于发现 77% 的鳞状细胞癌在额外的核心着丝粒基因中存在突变，我们还计划破坏选定的着丝粒基因并研究它们对表皮肿瘤发生的影响。因此，Aim II 将测试一个模型，在该模型中，突变的 Kinastrin 与离散的蛋白质组相互作用以促进肿瘤进展，并且在该模型中特定其他着丝粒基因的突变会导致癌变。 在拟议的资助结束后，我们计划进一步了解表皮肿瘤进展的机制，作为未来癌症预防和治疗策略的基础。