REGULATORS OF EPITHELIAL TUMOR PROGRESSION

上皮肿瘤进展的调节因子

• 批准号: 10620208
• 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: 2012
• 资助国家: 美国
• 起止时间: 2012-01-01 至 2028-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10620208
• 关键词: Ablation; Abnormal Cell; Acceleration; Address; Affect; COL7A1; Carcinoma; Cell Communication; Cell Surface Proteins; Cell secretion; Cell surface; Cells; Cessation of life; Collagen; Collagen Gene; Collagen Type XI; Communication; Complex; Data; Development; Dominant-Negative Mutation; Environment; Epidermis; Epidermolysis Bullosa; Epithelium; Extracellular Matrix; Extracellular Matrix Proteins; Eye; Frequencies; Funding; Future; Gene Expression; Gene Family; Genes; Glycine; Health; Human; Impairment; Induced Mutation; Inherited; Invaded; Knock-in; Link; Malignant Epithelial Cell; Malignant Neoplasms; Mediating; Messenger RNA; Methods; Middle East; Modeling; Morbidity - disease rate; Mutate; Mutation; Neoplasm Metastasis; Neoplasms; Normal tissue morphology; Oncogenic; Patients; Population; Proline; Protein Secretion; Proteins; Proteomics; RNA; Recurrence; Role; Series; Services; Set protein; Site; Skin; Southeastern Asia; Squamous cell carcinoma; Stromal Cells; Sun Exposure; Sunlight; Testing; Tissue Model; Tissues; Tumor Tissue; UV Mutagenesis; UV Radiation Exposure; Veterans; cancer cell; cancer invasiveness; cancer prevention; cancer therapy; cell community; cell stroma; cell type; early onset; extracellular; human tissue; in vivo; insight; interest; keratinocyte; military service; military veteran; mosaic; mutant; neoplastic; neoplastic cell; single-cell RNA sequencing; skin squamous cell carcinoma; sunlight-induced; therapeutic target; triple helix; tumor; tumor ablation; tumor progression; tumorigenesis

Epithelial tumor progression involves abnormal cell interactions with the extracellular environment as well abnormalities intrinsic to tumor cells themselves. Cutaneous squamous cell carcinoma (SCC) arises in skin from ultraviolet mutagenesis and disproportionately affects U.S. Veterans due to sun exposure incurred in military service. Nationally, SCC is the second most common cancer, with ~1 million new cases yearly. SCC can lead to disfigurement and, in rare cases, lethal metastases. This Merit Review recently identified collagens as the most highly mutated gene family in SCC, with the COL11A1 gene the second most frequently mutated gene overall in SCC (63%). It also found additional collagen genes mutated at high frequency (>20%) in SCC, including COL17A1 and COL4A4, as well as COL7A1, whose inherited mutation causes forms of epidermolysis bullosa with aggressive skin SCCs. SCC collagen mutations concentrate at prolines and glycines in the Gly-X-Y (with X and Y commonly proline) triple helical sequence, alterations known to produce dominant-negative collagens, raising the possibility that tumor-secreted mutant collagens may enable cancer in a trans-dominant fashion. Consistent with a pro-oncogenic function for mutant collagens, knocking in an SCC-associated COL11A1 mutation enhanced neoplastic invasion in vivo compared to isogenic COL11A1 wild-type control and ablating endogenously mutant COL11A1 in SCC tumors impaired tumorigenesis in vivo, indicating that mutant collagens functionally promote tumorigenesis. Aim I will test a model in which secreted mutant collagen proteins act in a non-cell autonomous fashion to accelerate tumorigenesis. First, it will determine if mutant COL11A1 can promote neoplastic progression of tumor cells in trans using mosaic tissue models. Second, it will quantify the impacts on epidermal tumor progression of mutations in multiple additional collagens that are frequently mutated in SCC, including COL17A1, COL4A4, and COL7A1. Aim I will study the action of COL11A1 in epidermal tumor progression and determine if other recurrently mutated collagens can also promote SCC. This project also recently used single-cell RNA-sequencing of 47,771 cells from a series of human SCC tumors, along with patient and site-matched normal control skin, to identify tumor cell subpopulations in SCC. This defined a new tumor-specific keratinocyte (TSK) population with no counterpart in normal tissue that expressed COL11A1 and other genes linked to cellular communication and invasion. Such tumor subpopulations enable neoplasia by communicating with each other through cell surface and secreted proteins, however, the sets of proteins – as opposed to RNAs – that are actually expressed in living tumors by specific tumor subpopulations have been a technical challenge to define. To address this, we developed a new proximity proteomics method, Secreted Protein Identification (SecrID) that can identify cell surface and secreted proteins within discrete cell populations of interest in heterogenous living tumor tissues in vivo to characterize cell subpopulation proteins in tumor progression. Aim II is based on a model in which the TSK SCC subpopulation uses specific cellular communication proteins to drive malignancy. First, it will use SecrID to define the secreted and cell surface proteins specific to the TSK cell subpopulation, based on the premise that these may mediate pro-neoplastic cellular communication. Second, it will ablate TSK subpopulation-specific genes, including those identified by SecrID, to define their impact on tumorigenesis and to search for new accessible therapeutic targets. Aim II will characterize the tumor-specific SCC TSK subpopulation and identify the molecules it uses to influence tumor progression. At the end of the proposed funding cycle, we plan to have characterized the actions of mutant collagens and newly defined tumor cell subpopulations in epidermal tumor progression.

上皮肿瘤进展涉及细胞与细胞外环境的异常相互作用，如 肿瘤细胞本身固有的异常。皮肤鳞状细胞癌 (SCC) 发生于 皮肤免受紫外线诱变，并且由于阳光照射而对美国退伍军人造成不成比例的影响 在服兵役。在全国范围内，鳞状细胞癌是第二大常见癌症，每年约有 100 万新发病例。 鳞状细胞癌可导致毁容，在极少数情况下，还会导致致命的转移。这项优点审查最近确定 胶原蛋白是鳞状细胞癌中突变程度最高的基因家族，其中 COL11A1 基因位居第二 SCC 中总体上经常发生突变的基因 (63%)。它还发现了额外的胶原蛋白基因在高位发生突变 SCC中出现频率（>20%），包括COL17A1和COL4A4，以及COL7A1，其遗传突变 引起大疱性表皮松解症并伴有侵袭性皮肤鳞状细胞癌。 SCC胶原蛋白突变集中于 Gly-X-Y（X 和 Y 通常为脯氨酸）三螺旋序列中的脯氨酸和甘氨酸、改变 已知会产生显性失活胶原蛋白，这增加了肿瘤分泌的突变胶原蛋白的可能性 可能以反式显性方式导致癌症。与突变体的促癌功能一致 胶原蛋白，敲入 SCC 相关的 COL11A1 突变可增强体内肿瘤侵袭 与 SCC 中的同基因 COL11A1 野生型对照和消除内源突变 COL11A1 相比 肿瘤损害体内肿瘤发生，表明突变胶原在功能上促进肿瘤发生。 目标我将测试一个模型，其中分泌的突变胶原蛋白以非细胞自主方式发挥作用 以加速肿瘤发生。首先，它将确定突变的 COL11A1 是否可以促进肿瘤进展 使用镶嵌组织模型反式分析肿瘤细胞。其次，它将量化对表皮肿瘤的影响 鳞状细胞癌中经常突变的多种其他胶原蛋白的突变进展，包括 COL17A1、COL4A4 和 COL7A1。目标我将研究 COL11A1 在表皮肿瘤进展中的作用 并确定其他经常突变的胶原蛋白是否也能促进鳞状细胞癌。 该项目最近还使用了来自一系列人类 SCC 的 47,771 个细胞的单细胞 RNA 测序 肿瘤以及患者和部位匹配的正常对照皮肤，以识别肿瘤细胞亚群 南卡罗来纳州。这定义了一种新的肿瘤特异性角质形成细胞（TSK）群体，在正常组织中没有对应物 表达 COL11A1 和其他与细胞通讯和入侵相关的基因。这样的肿瘤 亚群通过细胞表面相互通讯并分泌来实现肿瘤形成 然而，蛋白质是一组蛋白质（而不是 RNA），实际上在活体肿瘤中表达 特定肿瘤亚群的定义一直是一个技术挑战。为了解决这个问题，我们开发了 一种新的邻近蛋白质组学方法，分泌蛋白鉴定（SecrID），可以识别细胞表面 以及体内异质活体肿瘤组织中感兴趣的离散细胞群中的分泌蛋白 表征肿瘤进展中的细胞亚群蛋白。 Aim II 基于 TSK SCC 亚群使用特定细胞通信的模型 蛋白质驱动恶性肿瘤。首先，它将使用 SecrID 来定义分泌蛋白和细胞表面蛋白特异性 TSK细胞亚群，基于这些可能介导促肿瘤细胞的前提 沟通。其次，它将消除 TSK 亚群特异性基因，包括那些由 SecrID，以确定它们对肿瘤发生的影响并寻找新的可行的治疗靶点。目标二 将表征肿瘤特异性 SCC TSK 亚群并识别其用于影响的分子 肿瘤进展。 在拟议的资助周期结束时，我们计划表征突变胶原蛋白的作用 以及表皮肿瘤进展中新定义的肿瘤细胞亚群。