Creating a foundation for personalized age- and sex-based immune-targeted therapies from an ALS longitudinal cohort by identifying peripheral and central immune signatures

摘要 免疫系统有助于肌萎缩侧索硬化症（ALS）的进展和生存，以及治疗 靶向免疫系统的研究正受到越来越多的关注。然而，免疫系统的变化， ALS的病程和免疫功能的性别特异性改变需要更深入的分析， 开发个性化的ALS疗法和生物标志物。长期目标是利用免疫系统- tem减缓和阻止ALS进展的潜力。总体目标是确定 免疫特征、性别、年龄和性激素与神经元损伤、神经炎症和ALS进展相关- 锡永与生存。中心假设是外周免疫谱是一个重要的病理生理 ALS进展和生存的因素;性别、年龄和性激素水平影响这些特征;以及 对ALS患者特异性免疫谱的深入了解将产生新的药物靶点和治疗窗口。我们的- 将患者特异性免疫细胞谱与ALS进展和生存联系起来将有助于人- ALS的免疫调节治疗开发，并确定潜在的治疗窗口。中心- tral假说将追求三个目标：1）确定与ALS原相关的特异性免疫谱， 退化和存活率。2)评估性激素对ALS免疫特征、进展、 和生存;和3）确定免疫概况和相应的细胞途径，是最有毒的神经， 与中枢炎症相关的神经系统。In Aim 1外周血纵向免疫表型 来自ALS受试者的样品将产生复合免疫谱， 特征、进展和生存。目标2将确定观察到的性别依赖性关联是否- 免疫特征与ALS进展和存活之间是由性激素介导的， 可以改变免疫系统目标3将通过免疫特征聚类来丰富ALS受试者的队列;他们的外周血 将使用1）RNA-seq和2）细胞毒性研究，通过与以下物质共培养来分析所有免疫群体： iPSC衍生的神经元;受试者还将进行正电子发射断层扫描（PET）成像，以量化中枢神经系统的功能。 神经炎症数据集将被合成以构建预测模型并创建深度神经网络 能够将免疫特征与性别、年龄、疾病严重程度、进展和生存联系起来。 在申请人看来，所提出的是创新的，因为它严格审查了性别、年龄和 性激素水平对免疫细胞和ALS进展和生存的纵向研究。它也说明了 通过形成个体受试者的免疫特征来研究免疫细胞之间的相互作用， 与失调途径、细胞毒性和神经炎症相关的特定特征。拟议的重新- 搜索是重要的，因为它将提供关于不同免疫特征和相关途径的关键数据， 与ALS的进展和存活率相关，与性别、年龄和性激素有关。

ABSTRACT The immune system contributes to amyotrophic lateral sclerosis (ALS) progression and survival, and therapies to target the immune system are of burgeoning interest. However, the changes in the immune system during the course of ALS and the sex-specific alterations in immune function warrant a more in depth analysis in order to develop personalized ALS therapies and biomarkers. The long-term goals are to harness the immune sys- tem’s potential to slow and stop the progression of ALS. The overall objective is to determine how peripheral immune profiles, sex, age, and sex hormones, link to neuronal damage, neuroinflammation, and ALS progres- sion and survival. The central hypothesis is that peripheral immune profiles are an important pathophysiologic agent of ALS progression and survival; that sex, age and sex hormone levels impact these profiles; and that insight into ALS patient-specific immune profiles will yield new drug targets and therapeutic windows. Our ra- tionale is that linking patient-specific immune cell profiles to ALS progression and survival will facilitate person- alized immunomodulatory therapeutic development for ALS and identify potential treatment windows. The cen- tral hypothesis will be pursued with three aims: 1) Identify specific immune profiles that associate with ALS pro- gression and survival rates.; 2) Evaluate the effects of sex hormones on ALS immune profiles, progression, and survival; and 3) Identify immune profiles and corresponding cellular pathways that are most toxic to neu- rons and that associate with central inflammation. In Aim 1 longitudinal immunophenotyping of peripheral blood samples from ALS subjects will generate composite immune profiles that will then be linked to ALS subject characteristics, progression, and survival. Aim 2 will determine if observed sex-dependent associations be- tween immune profiles and ALS progression and survival are mediated by sex hormones, as sex hormones can alter immune profiles. Aim 3 will enrich a cohort of ALS subjects by immune profile clusters; their periph- eral immune populations will be analyzed using 1) RNA-seq and 2) cell toxicity studies via co-cultures with iPSC-derived neurons; subjects will also have positron emission tomography (PET) imaging to quantify central neuroinflammation. Datasets will be synthesized to build prediction models and create deep neural networks capable of associating immune profiles with sex, age, disease severity, progression, and survival The research proposed is innovative, in the applicants’ opinion, because it rigorously examines the effects of sex, age, and sex hormone levels on immune cells and ALS progression and survival in a longitudinal study. It also accounts for the interactions between immune cells by forming immune profiles for individual subjects and assesses how specific profiles associate with dysregulated pathways, cytotoxicity, and neuroinflammation. The proposed re- search is significant because it will provide critical data on the distinct immune profiles and pathways associ- ated with ALS progression and survival in a sex-, age- and sex hormone- specific fashion.