Role of Glycosylation of Dsg1 in Pemphigus acantholysis

描述(由申请人提供)： P.I.最近进入了皮肤自身免疫的研究领域，为其提供了强大的分子生物学背景。她的职业发展计划旨在获得自身免疫方面的高级培训，获得糖基化研究方面的研究经验，并提高动物模型研究的技术技能。这些新目标的实现，加上她过去的专业知识，将为候选人开发研究方向做好准备，解决糖基化在细胞黏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。她的研究计划探索多糖在天疱疮抗原/自身抗体相互作用中的作用。这一计划是建立在一个非常新的发现基础上的，即两种半乳糖特异性凝集素，JAXALIN和花生凝集素(PNA)，结合杆状病毒表达叶天疱疮(PF)抗原，桥粒芯糖蛋白1(RDsgl)，并保护小鼠免受PF IgG诱导的皮肤水泡。其他凝集素是无效的。假设通过空间位阻或通过诱导DSGL构象变化来干扰DSG1/自身抗体的相互作用，病原性抗DSGL抗体和凝集素竞争相同的DSG1表位也是可能的。目的1检测这些凝集素与哺乳动物表达的DSG1(糖基化程度可能不同于杆状病毒表达的DSG1)的相互作用。在目标2中，我们将选择性地去除DSG1的O-或N-糖链，然后测试它们与PF-Ig G和凝集素的结合能力。在目标3中，我们将定位DSG1上与凝集素和抗体结合的糖基化位点。在目标4中，我们将测试凝集素在体内是否与表皮结合，并阻止致病自身抗体的结合。我们将开发一种新的策略来测试凝集素是否可以防止现有病变的进展。这项资助的结果将有助于阐明PF的发病机制，并可能导致新的治疗方法。此外，产生的数据和获得的广泛知识将支持应聘者发展独立的职业生涯。

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.