Cockayne syndrome: role of the innate immune response in neurodegeneration

科凯恩综合征：先天免疫反应在神经退行性变中的作用

• 批准号: 8649097
• 负责人: ALAN M WEINER
• 金额: $ 15.3万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-05-01 至 2016-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8649097
• 关键词: Aging; Aicardi' s syndrome; Anti-Inflammatory Agents; Anti-inflammatory; Antiviral Agents; Antiviral Response; Autoantibodies; Autoimmune Responses; Biological; Biological Assay; Blood; Brain; C-terminal; Callithrix; Caring; Cells; Childhood; Chimeric Proteins; Clinical; Clinical Trials; Cockayne Syndrome; DNA Repair; Data; Defect; Development; Diagnosis; Diagnostic; Disease; Drug Formulations; ERCC3 gene; ERCC6 gene; Exhibits; Exons; Exploratory/Developmental Grant; Fibroblasts; Gene Expression; Gene Expression Profile; Genes; Genetic; Goals; Growth Factor; Health; Hearing; Heterogeneity; Human; Immune response; Immunosuppressive Agents; Infection; Inflammatory; Inflammatory Response; Interferons; International; Intervention; Investigation; Lead; Manuscripts; Messenger RNA; Methodology; Modality; Molecular; Mutation; Myelin; N-terminal; National Institute of General Medical Sciences; Nerve Degeneration; Normal Cell; Null Lymphocytes; Parents; Pathway interactions; Patients; Peripheral Blood Mononuclear Cell; Pharmaceutical Preparations; Primates; Progeria; Protein Array; Proteins; Publishing; Regulatory Pathway; Reporter; Research; Rewards; Role; Serum; Symptoms; Syndrome; Systemic Lupus Erythematosus; Testing; Transcription-Coupled Repair; Translational Research; Transposase; Virus Diseases; Work; base; calcification; chemokine; chromatin remodeling; clinical practice; cohort; cytokine; design; dysmyelination; high reward; high risk; leukodystrophy; mRNA Expression; meetings; novel; novel therapeutics; outcome forecast; protein expression; repository; research study; response

DESCRIPTION (provided by applicant): We propose to test an exciting new hypothesis regarding the causes and molecular mechanisms of a devastating childhood neurodevelopmental progeria known as Cockayne syndrome (CS). If our hypothesis is correct, it will open up new therapeutic options for this currently untreatable disease. CS has long been thought to be a disease of DNA repair because all 5 genes that can cause CS (CSA, CSB, XPB, XPD, and XPG) are essential for transcription-coupled nucleotide excision repair (TC-NER). Yet CS patients usually present with myelin defects and brain calcifications resembling those seen in Aicardi- Goutières syndrome (AGS), a childhood neurodevelopmental leukodystrophy caused by constitutive activation of an innate antiviral interferon response in the absence of viral infection. We recently discovered that the CSB gene, which causes over two thirds of all known cases of CS, contains a piggyBac transposon (PGBD3). As a result, the human CSB locus generates not one but two CSB-related proteins: functional CSB, and a CSB-PGBD3 fusion protein joining the N-terminal domain of CSB to a C-terminal PGBD3 transposase domain (Newman et al., 2008). Most surprisingly, we found that expression of the CSB-PGBD3 fusion protein in CSB-null cells induces a powerful interferon-like innate immune response, and that loss of CSB from normal cells also induces an innate antiviral response (Bailey et al., 2012). Our data suggest that the fusion protein may contribute to CS disease, and could provide an entry point for intervention to delay or ameliorate CS symptoms. We propose two complementary aims to determine whether CS patients display a similar interferon, antiviral, inflammatory, or autoimmune response: (Aim 1) Using Affymetrix microarrays, we will compare mRNA expression in peripheral blood mononuclear cells (PBMCs) of CS patients to our published data for CSB-null cells that stably express the CSB-PGBD3 fusion protein. Microarrays will enable us to detect an interferon, antiviral, or inflammatory gene expression signature, even if the genes are downregulated in patients by SOCS1 or related regulatory pathways. (Aim 2) Using Luminex bead-based assays and cell-based reporter assays, we will examine expression of interferons, cytokines, and chemokines at the protein level in CS serum and PBMCs. We will also use Invitrogen ProtoArray v5.0 protein arrays that display >9,000 human proteins to search for CS serum autoantibodies. Our proposal is high risk because the interferon-like response seen in CS cells may be muted in CS patients; however, the rewards could be high because confirmation of an interferon or autoimmune response would justify the use of immunomodulatory or anti-inflammatory biologicals and drugs for treating CS and related conditions.

描述（由申请人提供）：我们建议测试一个令人兴奋的新假设，该假设涉及被称为科凯恩综合征（CS）的破坏性儿童神经发育性早衰症的原因和分子机制。如果我们的假设是正确的，它将为这种目前无法治愈的疾病开辟新的治疗选择。长期以来，CS 一直被认为是一种 DNA 修复疾病，因为可引起 CS 的所有 5 个基因（CSA、CSB、XPB、XPD 和 XPG）对于转录偶联核苷酸切除修复 (TC-NER) 都是必需的。然而，CS 患者通常会出现髓磷脂缺陷和脑钙化，类似于 Aicardi-Goutières 综合征 (AGS)，这是一种儿童神经发育性脑白质营养不良，由先天抗病毒干扰素反应在没有病毒感染的情况下持续激活所致。我们最近发现，超过三分之二的已知 CS 病例均由 CSB 基因引起，该基因含有 PiggyBac 转座子 (PGBD3)。因此，人类 CSB 基因座产生的不是一种而是两种 CSB 相关蛋白：功能性 CSB 和将 CSB N 端结构域连接到 C 端 PGBD3 转座酶结构域的 CSB-PGBD3 融合蛋白（Newman 等人，2008）。最令人惊讶的是，我们发现CSB缺失细胞中CSB-PGBD3融合蛋白的表达会诱导强大的干扰素样先天免疫反应，而正常细胞中CSB的缺失也会诱导先天抗病毒反应（Bailey等人，2012）。我们的数据表明，融合蛋白可能导致 CS 疾病，并可以为延迟或改善 CS 症状的干预提供切入点。我们提出了两个互补的目标，以确定 CS 患者是否表现出类似的干扰素、抗病毒、炎症或自身免疫反应：（目标 1）使用 Affymetrix 微阵列，我们将 CS 患者外周血单核细胞 (PBMC) 中的 mRNA 表达与我们已发表的稳定表达 CSB-PGBD3 融合蛋白的 CSB-null 细胞的数据进行比较。微阵列将使我们能够检测干扰素、抗病毒或炎症基因表达特征，即使这些基因在患者中被 SOCS1 或相关调控途径下调。 （目标 2）使用基于 Luminex 微珠的测定和基于细胞的报告基因测定，我们将检查 CS 血清和 PBMC 中蛋白质水平的干扰素、细胞因子和趋化因子的表达。我们还将使用显示 > 9,000 种人类蛋白质的 Invitrogen ProtoArray v5.0 蛋白质阵列来搜索 CS 血清自身抗体。我们的建议是高风险的，因为在 CS 细胞中观察到的干扰素样反应在 CS 患者中可能会减弱；然而，回报可能很高，因为干扰素或自身免疫反应的确认将证明使用免疫调节或抗炎生物制剂和药物来治疗 CS 和相关病症是合理的。