Generating spatial and functional maps of cell-to-cell interactions in MS lesions

生成MS病变中细胞间相互作用的空间和功能图 单核RNA测序（sNuc-Seq）研究极大地促进了我们对 多发性硬化（MS）病变中的细胞异质性。然而，目前的CNS sNuc-Seq 方案选择性地对星形胶质细胞和小胶质细胞核进行采样不足，导致低分辨率的 种群异质性此外，sNuc-Seq不捕获空间信息，诸如 MS病变环境中细胞群的分布和相互作用。 在这里，我们提出使用一种新的方法来进行MS病变组织的sNuc-Seq，富集MS病变组织的sNuc-Seq。 未代表/罕见的细胞核，如星形胶质细胞、小胶质细胞和浸润性免疫细胞，通过Flow 分类我们将联合收割机与先进的计算工具相结合， 用于改善生物信号恢复和细胞信号分辨率的插补和批量校正 急性脱髓鞘、慢性活动性和髓鞘再生MS病变的异质性。我们将 利用亚群特异性转录组来计算预测 细胞群体之间基于受体-配体对表达的相互作用。我们将 进一步使用高度多重组织学成像方法定位细胞亚群 并证实受体-配体对在MS病变中的共定位。我们最终将在体外进行 研究阐明选定的受体-配体相互作用的功能影响。 我们的研究将确定组成亚群和受体配体相互作用组， 与急性脱髓鞘、慢性低度炎症和髓鞘再生相关。这些 相互作用或相互作用的组合可以靶向减少炎症或增强炎症反应。 此外，我们的数据集将提供丰富的资源， 疾病机制的功能研究。

Generating spatial and functional maps of cell-to-cell interactions in MS lesions Single nucleus RNA sequencing (sNuc-Seq) studies have greatly advanced our understanding of cellular heterogeneity in multiple sclerosis (MS) lesions. However, current CNS sNuc-Seq protocols selectively under-sample astrocyte and microglia nuclei, resulting in low resolution of population heterogeneity. Moreover, sNuc-Seq does not capture spatial information, such as distribution and interactions of cell populations within the MS lesion environment. Here, we propose to use a novel approach to sNuc-Seq of MS lesion tissue, enrichment for unrepresented/rare nuclei such as astrocytes, microglia and infiltrating immune cells through Flow sorting. We will combine this approach with advanced computational tools for denoising, imputation and batch correction to improve recovery of biological signal and resolution of cellular heterogeneity in acutely demyelinating, chronic active and remyelinating MS lesions. We will leverage the subpopulation-specific transcriptomes to computationally predict possible interactions between cell populations, based on expression of receptor-ligand pairs. We will further use a highly multiplexed histology imaging approach to localize cellular subpopulations and confirm co-localization of receptor-ligand pairs in MS lesions. We will finally conduct in vitro studies to elucidate the functional impact of selected receptor-ligand interactions. Our study will identify the constituent subpopulations and receptor-ligand interactomes that are associated with acute demyelination, chronic low-grade inflammation and remyelination. These interactions or combinations of interactions can be targeted to reduce inflammation or to enhance neurorepair in MS. Moreover, our datasets will provide a rich resource that will guide more functional studies on disease mechanisms.