Molecular understanding of self immunity in the brain

我最近开发了一种可编程噬菌体显示器，用于识别与人类副肿瘤神经系统疾病（PNDs）相关的致病性抗原。这个工作场所我在神经免疫学的迅速兴起的领域，因为pnd是衰弱中枢神经系统疾病，包括自身免疫性脑炎。PND患者体内含有针对脑抗原的自身抗体，自身抗体谱的特异性可以预测临床症状和预后。我描述了与pnd患者特异性相关的最大抗原集，其中一个是最近发表的新抗原KLHL11。在Aim 1中，我将描述额外的抗原并验证我的假设，即单个表位是PND患者自身免疫和脑炎的原因。K99可以让我掌握表位分析所需的先进生物信息学技能，这是导师实验室的优势。在Aim 2中，我测试了细胞内PND抗原是否通过非规范MHC-I类依赖途径引起自身免疫性脑炎，这是一种意想不到的机制，对治疗有影响。我将建立一个新的抗KLHL11 PND小鼠模型和一个互补的KLHL11敲除小鼠。结合现有免疫基因缺失的小鼠，我将使用免疫组织化学（IHC）和单细胞测序来测试PND脑内t细胞特异性靶向对非规范MHC途径的依赖性。K99将使我能够在课堂和指导老师的帮助下，获得目标2和目标3中所包含的必要的免疫学高级知识，并帮助指导最初的T细胞研究。此外，加州大学旧金山分校的使命，包括基础设施和研究环境，特别面向促进基础和临床免疫学家与神经学家之间的直接互动。我计划在这个目标过渡到R00阶段时继续利用这个专门的环境。在Aim 3中，我带来了我以前在小鼠遗传学和细胞神经生物学方面的专业知识，以及我的K99获得的分子免疫学专业知识，利用PND抗原（Aims 1和2）来生成大脑中靶向炎症的3D解剖图谱。我将测试脑区域特异性PND脑炎是否由中枢神经系统淋巴系统介导。在PND开始后的不同时间，免疫组化对T细胞进行脑清除，将提供入侵T细胞及其与抗原表达细胞关系的全面“3D”路线图。我所提出的研究将对决定神经元抗原如何引发大脑自身免疫反应的机制形成一个全面的图景，并假设T细胞迁移路线对解剖学上不同的自身免疫是特异性的，为识别不同PND病理背后的神经回路铺平道路，这是我作为一名独立研究者的目标。

I recently developed a programmable phage display for identifying pathogenic antigens related to human paraneoplastic neurological disorders (PNDs). This workplace me in the rapidly emerging area of neuro- immunology, as PNDs are debilitating central nervous system disorders that involve autoimmune encephalitis. Patients with PND harbor autoantibodies against brain antigens, and specificity in the autoantibody profile predicts clinical symptoms and prognosis. I have characterized the largest set of antigens that are linked specifically to patient PNDs, one of which, KLHL11, was novel and published recently. In Aim 1, I will characterize the additional antigens and test my hypothesis that a single epitope is causal to PND patient autoimmunity and encephalitis. The K99 will allow me to acquire the advanced bioinformatics skills necessary for epitope analysis, strengths of the mentor's lab. In Aim 2, I test whether intracellular PND antigens cause autoimmune encephalitis through a non-canonical MHC-I class-dependent pathway, an unexpected mechanism with implications for treatment. I will create a new mouse model for anti-KLHL11 PND and a complementary KLHL11 knockout mouse. Together with existing mice deleted in immunity genes, I will use immunohistochemistry (IHC) and single cell sequencing to test dependence in specific targeting of T-cells in the brain of PND on the non-canonical MHC pathway. The K99 will allow me, with classes and advisors, to acquire the necessary advanced knowledge in immunology built into Aims 2 and 3 and help guide the initial T cell studies. Further, the UCSF mission, including infrastructure and research environment, is geared particularly towards facilitating direct interactions between basic and clinical immunologists and neurologists. I plan to continue to exploit this specialized environment as I transition to the R00 phase in this aim. In Aim 3, I bring to bear my previous expertise in mouse genetics and cellular neurobiology, and my K99 acquired molecular immunology expertise, to leverage PND antigens (Aims 1 and 2) to generate 3D anatomical maps of targeted inflammation in the brain. I will test whether brain region-specific PND encephalitis is mediated by the CNS lymphatic system. IHC with brain clearing for T cells, at different times after initiation of PND, will provide a comprehensive “3D” roadmap of the invading T cells and their relationship to antigen-expressing cells. My proposed studies will result in a comprehensive picture of the mechanisms determining how neuronal antigens elicit a brain autoimmune response, and assuming T cell migration routes are specific for anatomically distinct auto immunities, pave the way for identifying neuronal circuitries underlying the distinct PND pathologies, my goal as an independent investigator.