miR-183/96/182 簇是常驻免疫细胞的检查点，可塑造角膜的细胞景观。
The miR-183/96/182 cluster is a checkpoint for resident immune cells and shapes the cellular landscape of the cornea.
机构信息
Predoctoral Training Program in Human Genetics, McKusick-Nathans Institute of Genetic Medicine, Department of Genetic Medicine, USA; Wilmer Eye Institute, School of Medicine, The Johns Hopkins University, Baltimore, MD, USA.
Genome Sciences Core, Wayne State University, Detroit, MI, USA.
出版信息
Ocul Surf. 2023 Oct;30:17-41. doi: 10.1016/j.jtos.2023.07.012. Epub 2023 Aug 2.
PURPOSE
The conserved miR-183/96/182 cluster (miR-183C) regulates both corneal sensory innervation and corneal resident immune cells (CRICs). This study is to uncover its role in CRICs and in shaping the corneal cellular landscape at a single-cell (sc) level.
METHODS
Corneas of naïve, young adult [2 and 6 months old (mo)], female miR-183C knockout (KO) mice and wild-type (WT) littermates were harvested and dissociated into single cells. Dead cells were removed using a Dead Cell Removal kit. CD45 CRICs were enriched by Magnetic Activated Cell Sorting (MACS). scRNA libraries were constructed and sequenced followed by comprehensive bioinformatic analyses.
RESULTS
The composition of major cell types of the cornea stays relatively stable in WT mice from 2 to 6 mo, however the compositions of subtypes of corneal cells shift with age. Inactivation of miR-183C disrupts the stability of the major cell-type composition and age-related transcriptomic shifts of subtypes of corneal cells. The diversity of CRICs is enhanced with age. Naïve mouse cornea contains previously-unrecognized resident fibrocytes and neutrophils. Resident macrophages (ResMφ) adopt cornea-specific function by expressing abundant extracellular matrix (ECM) and ECM organization-related genes. Naïve cornea is endowed with partially-differentiated proliferative ResMφ and contains microglia-like Mφ. Resident lymphocytes, including innate lymphoid cells (ILCs), NKT and γδT cells, are the major source of innate IL-17a. miR-183C limits the diversity and polarity of ResMφ.
CONCLUSION
miR-183C serves as a checkpoint for CRICs and imposes a global regulation of the cellular landscape of the cornea.
目的
保守的 miR-183/96/182 簇（miR-183C）调节角膜感觉神经支配和角膜固有免疫细胞（CRIC）。本研究旨在揭示其在 CRIC 中的作用以及在单细胞（sc）水平上塑造角膜细胞景观的作用。
方法
采集新生、年轻成年[2 个月和 6 个月（mo）]、雌性 miR-183C 敲除（KO）小鼠和野生型（WT）同窝小鼠的角膜，将其解离成单细胞。使用死细胞去除试剂盒去除死细胞。通过磁激活细胞分选（MACS）富集 CD45 CRIC。构建 scRNA 文库并进行测序，然后进行全面的生物信息学分析。
结果
WT 小鼠角膜的主要细胞类型组成在 2 至 6 mo 时相对稳定，然而角膜细胞亚类的组成随年龄而变化。miR-183C 的失活破坏了主要细胞类型组成的稳定性和角膜细胞亚类与年龄相关的转录组变化。CRIC 的多样性随年龄增长而增强。新生小鼠角膜中含有以前未被识别的固有纤维母细胞和中性粒细胞。固有巨噬细胞（ResMφ）通过表达丰富的细胞外基质（ECM）和 ECM 组织相关基因而获得角膜特异性功能。新生角膜具有部分分化的增殖性 ResMφ，并包含小胶质细胞样 Mφ。固有淋巴细胞，包括先天淋巴样细胞（ILCs）、NKT 和 γδT 细胞，是先天 IL-17a 的主要来源。miR-183C 限制了 ResMφ 的多样性和极性。
结论
miR-183C 是 CRIC 的检查点，对角膜细胞景观具有全局调节作用。
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