REGULATORS OF EPITHELIAL TUMOR PROGRESSION

上皮肿瘤进展的调节因子

• 批准号: 9891608
• 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 2024-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9891608
• 关键词: Abnormal Cell; Address; Affect; Carcinoma; Cell Communication; Cell Surface Proteins; Cell surface; Cells; Cessation of life; Collagen; Collagen Gene; Collagen Type XI; Communication; Complex; DNA Sequence Alteration; Data; Development; Dominant-Negative Mutation; Environment; Epidermis; Epidermolysis Bullosa; Epithelial; 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; Lead; Link; Malignant Epithelial Cell; Malignant Neoplasms; Mediating; Messenger RNA; Methods; Middle East; Military Personnel; Modeling; Morbidity - disease rate; Mutate; Mutation; Neoplasm Metastasis; Neoplasms; Normal tissue morphology; Oncogenic; Patients; Population; Proline; 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; base; cancer cell; cancer invasiveness; cancer prevention; cancer therapy; cell community; cell stroma; cell type; early onset; extracellular; human tissue; in vivo; insight; interest; keratinocyte; mutant; neoplastic; neoplastic cell; single-cell RNA sequencing; skin squamous cell carcinoma; sunlight-induced; therapeutic target; tissue mosaicism; tumor; 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.

上皮肿瘤进展涉及与细胞外环境异常的相互作用 肿瘤细胞本身的固有异常。皮肤鳞状细胞癌（SCC）出现 紫外线发生的皮肤和由于阳光暴露而对美国退伍军人的影响不成比例 服兵役。在全国范围内，SCC是第二大常见的癌症，每年约有100万例新病例。 SCC可能导致毁容，在极少数情况下会导致致命转移。该优点评论最近确定 胶原蛋白是SCC中最高度突变的基因家族，col11a1基因第二大基因 经常在SCC中总体突变基因（63％）。它还发现了在高处突变的其他胶原基因 SCC中的频率（> 20％），包括COL17A1和COL4A4以及COL7A1，其遗传突变 引起具有侵略性皮肤SCC的表皮溶液的形式。 SCC胶原蛋白突变集中在 Gly-X-Y（X和Y通常是脯氨酸）三重螺旋序列中的脯氨酸和甘氨酸，改变 已知会产生显性阴性胶原蛋白，从而提高了肿瘤分泌突变胶原的可能性 可以以跨性别的方式使癌症造成癌症。与突变体的促进功能一致 胶原蛋白，敲打与SCC相关的COL11A1突变增强了体内的肿瘤入侵 与SCC中的Isenic Col11a1野生型对照和消融内源性突变体Col11a1相比 肿瘤在体内受损的肿瘤发生障碍，表明突变体胶原蛋白在功能上促进了肿瘤发生。 目的，我将测试一个模型，其中分泌的突变胶原蛋白以非细胞自动方式起作用 加速肿瘤发生。首先，它将确定突变col11a1是否可以促进肿瘤进展 使用镶嵌组织模型在反式中的肿瘤细胞。其次，它将量化对表皮肿瘤的影响 在SCC中经常突变的多个其他胶原蛋白中突变的进展，包括 COL17A1，COL4A4和COL7A1。目的我将研究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可以促进SCC。 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 肿瘤以及患者和现场匹配的正常对照皮肤，以识别肿瘤细胞亚群 SCC。这定义了新的肿瘤特异性角质形成细胞（TSK）种群，在正常组织中没有对应物 这表达了与细胞通信和侵袭有关的Col11a1和其他基因。这样的肿瘤 亚群通过通过细胞表面相互通信并分泌 然而，蛋白质（与RNA相反）的蛋白质套件实际上在生物肿瘤中表达 通过特定的肿瘤亚群是定义的技术挑战。为了解决这个问题，我们开发了 一种新的接近蛋白质组学方法，分泌的蛋白质鉴定（SECRID），可以鉴定细胞表面 并在体内异质活肿瘤组织中离散感兴趣的细胞群体内的分泌蛋白 表征肿瘤进展中的细胞亚群蛋白。 AIM II基于一个模型，TSK SCC亚群使用特定的蜂窝通信 蛋白质驱动恶性肿瘤。首先，它将使用Secred ID来定义特定的分泌和细胞表面蛋白 基于以下前提，这些前提可能介导前塑性细胞 沟通。其次，它将消除TSK亚群特异性基因，包括由 SECRID，以定义其对肿瘤发生的影响并寻找新的可访问治疗靶标。目标II 将表征肿瘤特异性的SCC TSK亚群，并确定其影响的分子 肿瘤进展。 在拟议的资金周期结束时，我们计划表征突变胶原的作用 以及新定义的表皮肿瘤进展中的肿瘤细胞亚群。