Transcriptome dynamics during vocal fold injury and repair

摘要 声带(VF)疤痕是声带手术后声音不佳的最大原因，可以有 对生活质量有显著的负面影响。VF瘢痕形成的特征是细胞外的组织紊乱。 固有层的基质(ECM)以及周围上皮和血管的完整性改变 VF柔韧性的总体降低。没有一种治疗方法可以消除形成的疤痕或逆转疤痕 队形。由于缺乏对声带瘢痕形成的了解，针对声带疤痕的研究一直受到限制。 在损伤空间的背景下，贡献细胞亚群和细胞状态动态。的总目标是 这项工作是为了弄清参与室颤损伤反应的细胞的亚群、位置和邻近程度。 并在体内修复。目标1将使用单细胞RNA测序分析来识别细胞亚群 在损伤和愈合过程中存在，以及在和之间差异表达的基因 以确定转录组中哪些特定细胞的变化是重要的。目标2将调查空间 利用空间RNA测序分析进行差异表达以确定富含不同基因集的生态位 并在伤口愈合和修复过程中，在空间和时间上将细胞亚群定位于室颤。中环 这一指导性研究培训方案的假设是，存在免疫细胞亚群， 对损伤有反应的成纤维细胞和上皮细胞，以及这些细胞的空间位置和相互之间的距离 其他基因对协调基因的表达有显著影响。这项提案将确定参与的小区 在炎症过程中，将基因表达的亚群和定位定位到室颤中的小生境， 创面愈合的增殖期、重建期和成熟期疤痕提供靶向再生信息 接近了。

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.