Molecular profiling of resident and infiltrating mononuclear phagocytes during rapid adult retinal degeneration using single-cell RNA sequencing
Molecular profiling of resident and infiltrating mononuclear phagocytes during rapid adult retinal degeneration using single-cell RNA sequencing
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DOI：
10.1038/s41598-019-41141-0
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发表时间：
2019-03-19
影响因子：
4.6
通讯作者：
Burns, Marie E.
中科院分区：
文献类型：
--
作者：
Ronning, Kaitryn E.;Kaden, Sarah J.;Burns, Marie E.
神经炎症通常伴随着神经退行性变，但是在退行性变期间驻留和浸润免疫细胞的具体作用仍然存在争议。在疾病进展期间评估髓样细胞特异性功能的大部分困难来自于不能清楚地区分处于中枢神经系统内分化和活化的各个阶段的活化的小胶质细胞和骨髓来源的单核细胞和巨噬细胞。使用感光细胞死亡的诱导模型，我们研究了小鼠视网膜变性开始后浸润单核细胞和巨噬细胞亚群的患病率。体内视网膜成像显示，在光感受器变性的48小时内，CCR2（+）白细胞穿过视网膜血管浸润并进入实质。免疫组织化学和流式细胞术证实并鉴定这些白细胞为CD11b（+）CD45（+）细胞。整个CD11b（+）CD45（+）群体的单细胞mRNA测序显示存在静息小胶质细胞、活化小胶质细胞、单核细胞和巨噬细胞以及这四种主要细胞类别中的12个不同亚群。我们的研究结果表明，中枢神经系统内细胞自主变性的先天免疫反应具有不可估量的分子异质性，并强调了无偏见的高通量和高维分子技术（如scRNAseq）的必要性，以了解疾病进展期间免疫应答者的复杂性和不断变化的景观。
Neuroinflammation commonly accompanies neurodegeneration, but the specific roles of resident and infiltrating immune cells during degeneration remains controversial. Much of the difficulty in assessing myeloid cell-specific functions during disease progression arises from the inability to clearly distinguish between activated microglia and bone marrow-derived monocytes and macrophages in various stages of differentiation and activation within the central nervous system. Using an inducible model of photoreceptor cell death, we investigated the prevalence of infiltrating monocytes and macrophage subpopulations after the initiation of degeneration in the mouse retina. In vivo retinal imaging revealed infiltration of CCR2(+) leukocytes across retinal vessels and into the parenchyma within 48 hours of photoreceptor degeneration. Immunohistochemistry and flow cytometry confirmed and characterized these leukocytes as CD11b(+)CD45(+) cells. Single-cell mRNA sequencing of the entire CD11b(+)CD45(+) population revealed the presence of resting microglia, activated microglia, monocytes, and macrophages as well as 12 distinct subpopulations within these four major cell classes. Our results demonstrate a previously immeasurable degree of molecular heterogeneity in the innate immune response to cell-autonomous degeneration within the central nervous system and highlight the necessity of unbiased high-throughput and high-dimensional molecular techniques like scRNAseq to understand the complex and changing landscape of immune responders during disease progression.