Role of Glycosylation of Dsg1 in Pemphigus acantholysis

Dsg1 糖基化在棘层松解型天疱疮中的作用

• 批准号: 6970388
• 负责人: NING LI
• 金额: $ 12.13万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-09-16 至 2010-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6970388
• 关键词: antigen antibody reaction; autoantibody; autoantigens; cadherins; clinical research; conformation; cytoprotection; glycosylation; human tissue; laboratory mouse; lectin; molecular pathology; pemphigus; protein binding; protein structure function; receptor binding; skin disorder chemotherapy

DESCRIPTION (provided by applicant): The P.I. has recently entered the research field of cutaneous autoimmunity bringing a strong molecular biology background. Her career development plan is designed to secure advanced training in autoimmunity, gain research experience in glycosylation study, and enhance technical skills in animal model study. Achievement of these new objectives together with her past expertise will prepare the candidate to develop a research direction, addressing role of glycosylation in cell-adhesion and skin blistering diseases. Under the guidance of Dr. L.A. Diaz, her mentor, she will interact closely with co-mentors/consultants during the proposed research. Her research plan explores the role of glycans on pemphigus antigen/auto antibody interactions. This plan was build upon a very novel finding that two galactose-specific lectins, jacalin and peanut agglutinin (PNA), bind baculovirus expressed pemphigus foliaceus (PF) antigen, desmoglein 1 (rDsgl), and protect mice from PF IgG-induced skin blisters. Other lectins are ineffective. It is hypothesized that binding of jacalin or PNA to Dsg1 interfere the Dsg1/autoantibody interaction by steric hindrance or by inducing conformational changes on Dsgl It is also feasible that pathogenic anti-Dsgl antibodies and lectins compete for the same Dsg1 epitopes. Aim 1 tests the interaction of these lectins with mammalian-expressed Dsg1 (which may be glycosylated differently than the baculovirus-expressed Dsg1). In Aim 2, we will selectively remove the O- or N-glycans of Dsg1 and then test their binding ability to PF IgG and the lectins. In Aim 3, we will map the glycosylation sites on Dsg1 that are bound by lectins and antibodies. In Aim 4, we will test whether lectins bind epidermis in vivo and prevent the binding of pathogenic autoantibodies. We will develop a new strategy to test whether the lectins can prevent progression of existing lesions. The outcome of this grant will shed light on the pathogenesis of PF and may lead to novel therapies. Additionally, the data generated and broad knowledge gained will support the candidate to develop an independent career.

描述（由申请人提供）： P.I.最近进入皮肤自身免疫研究领域，带来了强大的分子生物学背景。她的职业发展计划旨在确保自身免疫方面的高级培训，获得糖基化研究的研究经验，并提高动物模型研究的技术技能。这些新目标的实现以及她过去的专业知识将为候选人制定一个研究方向做好准备，解决糖基化在细胞粘附和皮肤起泡疾病中的作用。在她的导师 L.A. Diaz 博士的指导下，她将在拟议的研究期间与合作导师/顾问密切互动。她的研究计划探讨聚糖对天疱疮抗原/自身抗体相互作用的作用。该计划基于一项非常新颖的发现，即两种半乳糖特异性凝集素（jacalin 和花生凝集素 (PNA)）可结合杆状病毒表达的落叶型天疱疮 (PF) 抗原桥粒芯糖蛋白 1 (rDsgl)，并保护小鼠免受 PF IgG 诱导的皮肤水疱。其他凝集素无效。假设jacalin或PNA与Dsg1的结合通过空间位阻或通过诱导Dsgl上的构象变化来干扰Dsg1/自身抗体相互作用。致病性抗Dsgl抗体和凝集素竞争相同的Dsg1表位也是可行的。目标 1 测试这些凝集素与哺乳动物表达的 Dsg1（其糖基化可能与杆状病毒表达的 Dsg1 不同）的相互作用。在目标 2 中，我们将选择性去除 Dsg1 的 O-或 N-聚糖，然后测试它们与 PF IgG 和凝集素的结合能力。在目标 3 中，我们将绘制 Dsg1 上与凝集素和抗体结合的糖基化位点图谱。在目标4中，我们将测试凝集素是否在体内结合表皮并阻止致病性自身抗体的结合。我们将开发一种新策略来测试凝集素是否可以阻止现有病变的进展。这笔资助的结果将揭示 PF 的发病机制，并可能带来新的治疗方法。此外，生成的数据和获得的广泛知识将支持候选人发展独立的职业生涯。