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 针对免疫系统的研究正引起人们的极大兴趣。然而，免疫系统在治疗过程中的变化 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，以便 开发个性化的肌萎缩侧索硬化症疗法和生物标记物。长期目标是利用免疫系统- 延缓和阻止肌萎缩侧索硬化症进展的潜力。总体目标是确定外围设备 免疫档案、性别、年龄和性激素，与神经元损伤、神经炎症和肌萎缩侧索硬化症的进展有关- 锡安和生存。中心假设是外周免疫模式是一种重要的病理生理机制。 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 洞察ALS患者特定的免疫特征将产生新的药物靶点和治疗窗口。我们的RA- 将患者特定的免疫细胞图谱与ALS的进展和生存联系起来，将有助于个人- 肌萎缩侧索硬化症的免疫调节治疗进展和确定潜在的治疗窗口。该中心- TRAL假说将以三个目标进行：1)识别与ALS PRO相关的特定免疫图谱。 评估性激素对肌萎缩侧索硬化症免疫状态、进展、 和存活率；以及3)确定对神经元毒性最大的免疫图谱和相应的细胞通路。 以及与中枢性炎症有关的因子。目标1外周血纵向免疫表型分析 来自ALS受试者的样本将生成复合免疫配置文件，然后将其链接到ALS受试者 特征、进展和生存。目标2将确定观察到的性别依赖关联是否- 吐温免疫模式和肌萎缩侧索硬化症的进展和存活是由性激素作为性激素来调节的 可以改变免疫模式。AIM 3将通过免疫图谱簇丰富ALS受试者队列；他们的外周- 外周免疫群体将通过1)rna-seq和2)细胞毒性研究通过与 IPSC来源的神经元；受试者还将进行正电子发射断层扫描(PET)成像，以量化中枢 神经炎。将综合数据集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 在申请者看来，该建议是创新的，因为它严格地检查了性别、年龄和 性激素水平对免疫细胞和肌萎缩侧索硬化症进展和生存的纵向研究。它还解释了 通过形成单个受试者的免疫配置文件来研究免疫细胞之间的相互作用，并评估如何 特定的特征与失调的通路、细胞毒性和神经炎症有关。建议的重行- 搜索具有重要意义，因为它将提供有关不同免疫图谱和相关途径的关键数据。 肌萎缩侧索硬化症的进展和生存以性别、年龄和性激素特有的方式进行。

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.