Developing in situ transcriptomics of a bioprinted follicular skin model

项目摘要/摘要 这项提议的目标是开发一个平台来模拟皮肤中宿主和微生物的相互作用，具有3D 生物印染毛囊皮肤模型和宿主细胞和微生物细胞的空间转录以保持生理 相关互动。在人类皮肤中，大多数微生物与宿主皮肤上皮和皮肤 免疫被认为发生在更深的保护性附属器结构中，例如毛囊和腺体。为 例如，最近在老鼠身上的研究表明，不同组织驻留细胞之间存在动态平衡相互作用， 皮脂腺和微生物组。考虑到包括微生物组的大量多样性 数百种不同的细菌和真菌物种，对这种潜力进行建模将是非常有影响力的 系统地进行互动。此外，研究与人类细胞的相互作用将最好地提供 关于细胞对微生物定植的反应的翻译数据。然而，3D技术有很大的局限性 现有的皮肤模型--它们缺乏皮肤微生物所在的复杂结构，第二，目前 研究寄主-微生物相互作用的方法缺乏对空间活动的分辨率。在这里，我们的两个目标 解决这些关键限制。在目标1中，我们将开发包含毛囊的3D生物打印皮肤组织模型 结构。在目标2中，我们将开发用于微生物空间图谱的原位微生物测序技术 这些组织和其他可用的人体3D皮肤模型的定植。这些目标加在一起将使 我们将解决皮肤微生物组潜在生物学的基本问题，我们将在 这辆R21。我们的成功将带来两项新技术，这将使构建高分辨率 微生物组和宿主相互作用的空间图，使基础研究成为可能 菌株间的相互作用和毒力调节，以促进皮肤动态平衡。

PROJECT SUMMARY / ABSTRACT The goal of this proposal is to develop a platform to model host-microbe interactions in skin, featuring a 3D bioprinted follicular skin model and spatial transcriptomics of host and microbial cells to retain physiologically relevant interactions. In human skin, a majority of microbial interactions with the host epithelium and cutaneous immunity is thought to occur in deeper, protective adnexal structures, e.g., hair follicles and glands. For example, recent work in mice has suggested a homeostatic interaction between different tissue resident cells, sebaceous glands, and the microbiome. Given the substantial diversity of the microbiome – encompassing hundreds of different bacterial and fungal species, it would be highly impactful to model such potential interactions systematically. In addition, examining interactions with human cells would best provide translational data on cellular response to microbial colonization. However, there are major limitations in the 3D skin models that are available – they lack complex structures where skin microbes reside, and second, current approaches to investigate host-microbiome interactions lack resolution on spatial activity. Here, our two aims address these key limitations. In Aim 1, we will develop a 3D bioprinted skin tissue model containing follicular structures. In Aim 2, we will develop in situ microbial sequencing technology for spatial profiling of microbial colonization of these tissues and other available human 3D skin models. These aims taken together will allow us to address fundamental questions on the underlying biology of the skin microbiome, which we will pilot in this R21. Our success would result in two new technologies that would enable construction of a high-resolution spatial map of the microbiome and host interactions, enabling foundational investigations into mechanisms of interstrain interactions and regulation of virulence to promote skin homeostasis.