Transcriptome dynamics during vocal fold injury and repair

摘要 声带（VF）疤痕是声带手术后声音差的最大原因， 对生活质量产生重大负面影响。VF瘢痕形成的特征是细胞外基质的紊乱， 基质（ECM）的固有层，以及周围上皮细胞的完整性的改变， VF柔韧性的总体降低。没有治疗方法可以消除已形成的疤痕或逆转疤痕 阵由于缺乏对声带瘢痕形成的了解， 在损伤空间的背景下，贡献细胞亚群和细胞状态动态。的总目标 这项工作是为了洗脱有助于VF损伤反应的细胞亚群、位置和邻近性 并在体内修复。目的1将使用单细胞RNA测序分析来鉴定细胞亚群 在损伤和愈合过程中存在，以及基因之间和之间的差异表达（DE） 它们来确定转录组中细胞特异性变化的重要性。目标2将研究空间 使用空间RNA测序分析鉴定富集不同基因组的小生境的差异表达 并在伤口愈合和修复期间在空间和时间上将细胞亚群定位于VF。中央 这种指导性研究培训建议的假设是存在免疫细胞亚群， 成纤维细胞和上皮细胞对损伤做出反应，并且这些细胞的空间位置和与每个细胞的接近程度 其他对协调基因表达有显著影响。该提案将确定参与细胞 亚群，并将基因表达位置同一性映射到炎症期间VF中的小生境， 伤口愈合的增殖、重塑和成熟瘢痕阶段，以告知靶向再生 接近。

ABSTRACT Vocal fold (VF) scarring is the single greatest cause of poor voice after vocal fold surgery and can have a significant negative impact on quality of life. VF scarring is characterized by disorganization in the extracellular matrix (ECM) of the lamina propria as well as alterations in the integrity of the surrounding epithelium and an overall reduction in VF pliability. There is no treatment that can eliminate formed scars or reverse scar formation. Research specific to vocal fold scarring has been limited by a lack of understanding of the contributing subpopulations of cells and cell state dynamism in the context of injury space. The overall goal of this work is to eluicate subpopulations, location, and proximity of cells that contribute to the VF injury response and repair in vivo. Aim 1 will use single cell RNA sequencing analysis to identify the subpopulations of cells present during injury and healing, as well as genes that are differentially expressed (DE) between and among them to determine where cell-specific changes in the transcriptome are important. Aim 2 will investigate spatial differential expression using spatial RNA sequencing analysis to identify niches enriched for distinct gene sets and localize cell subpopulations to the VF in space and time during wound healing and repair. The central hypothesis of this mentored research training proposal is that there are subpopulations of cells – immune, fibroblasts and epithelial cells that respond to injury, and that these cells’ spatial location and proximity to each other have a significant effect on coordinated gene expression. This proposal will identify participating cell subpopulations and map gene expression positional identities to niches in the VF during inflammatory, proliferative, remodeling and mature scar phases of wound healing to inform targeted regenerative approaches.