Autoinflammation in Human Melanoma

人类黑色素瘤的自身炎症

• 批准号: 8334977
• 负责人: Mayumi Fujita
• 金额: --
• 依托单位: VA EASTERN COLORADO HEALTH CARE SYSTEM
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-07-01 至 2016-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8334977
• 关键词: Acute; Address; Age; Antineoplastic Agents; Biological; Biological Process; Cancer Biology; Cancer Patient; Cell Line; Cells; Clinical; Clinical Oncology; Coculture Techniques; Complex; Exhibits; Genes; Genetically Engineered Mouse; Human; Immunocompetent; Implant; Incidence; Individual; Inflammation; Inflammatory; Interleukin-1; Lead; Life; Link; Malignant Neoplasms; Measures; Melanoma Cell; Metastatic Melanoma; Military Personnel; Modeling; Molecular; Molecular Target; Mus; Nature; Neoplasms; Pathway interactions; Patients; Play; Population; Prevention; Production; Property; Receptor Signaling; Recruitment Activity; Recurrence; Reporting; Risk; Role; Services; Signal Transduction; Stimulus; Stromal Cells; Sun Exposure; Sunburn; Tissues; Tumor Biology; Xenograft Model; aldehyde dehydrogenases; anticancer research; cancer stem cell; cytokine; drug development; effective therapy; melanoma; mouse model; neoplastic cell; novel; outcome forecast; protein complex; small hairpin RNA; tumor; tumor growth; tumor initiation; tumor progression; tumor xenograft; young woman

DESCRIPTION (provided by applicant): Melanoma remains a challenge in clinical oncology. Evidence increasingly suggests that inflammation can be associated with unfavorable clinical prognosis in cancer patients. IL-1b is a pleiotropic pro-inflammatory cytokine and induces a broad portfolio of genes from many cells. We have shown that metastatic melanoma cells spontaneously secrete biologically active IL-1b in the absence of exogenous stimuli because of constitutive activation of IL-1 receptor (IL-1R) signaling and a multi-protein complex, "inflammasome", exhibiting a feature of "auto-inflammation". We hypothesize that auto-inflammation from melanoma cells contributes to tumor growth and progression in tumor microenvironment. We propose to unravel the molecular (in Aim 1) and biological (in Aim 2) mechanisms of auto-inflammation in human melanoma cells. In Aim 3, we will investigate if auto-inflammation is associated with a subpopulation of melanoma cells such as cancer stem cells (CSCs). The specific aims are: Specific Aim 1. To analyze molecular mechanisms regulating auto-inflammation in human melanoma. We will delineate molecular mechanisms of auto-inflammation by assessing the role of inflammasomes in Aim 1.1. ASC, NLRP1 and NLRP3 will be silenced from human metastatic melanoma cell lines with short hairpin RNA and its role on molecular signaling and biological function will be assessed. We will then examine inflammasome-independent pathway in Aim 1.2. Specific Aim 2. To examine the biological role of auto-inflammation in tumor microenvironment. We will utilize a direct patient tumor xenograft model by implanting human melanoma tissues directly into mice in Aim 2.1. This is one of the best models in human cancer research that recapitulate complex tumor microenvironment. In addition, a genetically engineered mouse model (GEMM) of melanoma will be used in Aim 2.2 to better address tumor biology in immunocompetent syngeneic mice. We will examine the role of melanoma-derived IL-1b and auto-inflammation on survival of melanoma cells and recruiting and activating stromal cells in tumor microenvironment. Specific Aim 3. To assess if auto-inflammation is associated with a subpopulation of melanoma cells such as CSCs. We reported heterogeneous IL-1b expression in metastatic melanoma tumors. The CSC hypothesis proposes that a subpopulation of tumor cells is responsible for survival and maintaining of the growth of neoplastic tissue. Recently, we identified a subpopulation of human melanoma cells that possess CSC properties and express high aldehyde dehydrogenase (ALDH) activity. These cells express unique genes and IL-1 signaling network, suggesting that melanoma auto-inflammation may be associated with CSCs. We will measure IL-1b production and secretion from 4 potential CSCs and non-CSCs (Aim 3.1) and assess autoinflammatory nature of CSCs on non-CSCs by co-culturing these populations in Aim 3.2. The subpopulation that shows auto-inflammation will be further analyzed phenotypically and biologically in Aim 3.3. Both human tumors and GEMM tumors will be used. Until now, little has been studied on the mechanisms and roles of inflammasome and auto-inflammation from non-immunological cells such as tumor cells. Since IL-1b plays an important role in cancer biology, elucidating biological mechanisms of auto-inflammation in cancer will help our understanding of tumor initiation, progression and recurrence and is critical for a successful treatment against melanoma. It will also lead to the discovery of novel molecular targets for cancer drug development.

描述（由申请人提供）： 黑色素瘤仍然是临床肿瘤学的一个挑战。越来越多的证据表明，炎症可能与癌症患者不良的临床预后有关。 IL-1b 是一种多效性促炎细胞因子，可诱导许多细胞产生广泛的基因组合。我们已经证明，由于 IL-1 受体 (IL-1R) 信号传导和多蛋白复合物“炎性体”的组成型激活，转移性黑色素瘤细胞在没有外源刺激的情况下自发分泌具有生物活性的 IL-1b，表现出以下特征： “自身炎症”。我们假设黑色素瘤细胞的自身炎症有助于肿瘤微环境中的肿瘤生长和进展。我们建议揭示人类黑色素瘤细胞自身炎症的分子（目标 1）和生物（目标 2）机制。在目标 3 中，我们将研究自身炎症是否与癌症干细胞 (CSC) 等黑色素瘤细胞亚群相关。具体目标是： 具体目标1.分析调节人类黑色素瘤自身炎症的分子机制。我们将通过评估目标 1.1 中炎症小体的作用来描述自身炎症的分子机制。具有短发夹 RNA 的人转移性黑色素瘤细胞系中的 ASC、NLRP1 和 NLRP3 将被沉默，并评估其在分子信号传导和生物学功能中的作用。然后我们将在目标 1.2 中检查独立于炎症小体的途径。具体目标2.研究肿瘤微环境中自身炎症的生物学作用。在目标 2.1 中，我们将通过将人类黑色素瘤组织直接植入小鼠体内来利用直接患者肿瘤异种移植模型。这是人类癌症研究中再现复杂肿瘤微环境的最佳模型之一。此外，Aim 2.2 将使用黑色素瘤基因工程小鼠模型 (GEMM)，以更好地解决具有免疫能力的同基因小鼠的肿瘤生物学问题。我们将研究黑色素瘤衍生的 IL-1b 和自身炎症对黑色素瘤细胞存活以及在肿瘤微环境中募集和激活基质细胞的作用。具体目标 3. 评估自身炎症是否与黑色素瘤细胞亚群（如 CSC）相关。我们报道了转移性黑色素瘤中 IL-1b 的异质表达。 CSC 假说提出，肿瘤细胞亚群负责肿瘤组织的存活和维持生长。最近，我们鉴定了一个具有 CSC 特性并表达高乙醛脱氢酶 (ALDH) 活性的人类黑色素瘤细胞亚群。这些细胞表达独特的基因和 IL-1 信号网络，表明黑色素瘤自身炎症可能与 CSC 相关。我们将测量 4 个潜在 CSC 和非 CSC 的 IL-1b 产生和分泌（目标 3.1），并通过在目标 3.2 中共培养这些群体来评估非 CSC 上 CSC 的自身炎症性质。显示自身炎症的亚群将在目标 3.3 中进一步进行表型和生物学分析。人类肿瘤和 GEMM 肿瘤都将被使用。到目前为止，关于炎症小体和非免疫细胞（例如肿瘤细胞）自身炎症的机制和作用的研究还很少。由于 IL-1b 在癌症生物学中发挥着重要作用，阐明癌症中自身炎症的生物学机制将有助于我们了解肿瘤的发生、进展和复发，对于成功治疗黑色素瘤至关重要。它还将导致癌症药物开发新分子靶标的发现。