Integrating tissue engineering and microfluidics to model the spatial niches of the human endometrium in vitro with guidance from in vivo multiomics data

子宫内膜--子宫最里面的粘膜层，提供胚胎的位置 植入-是至关重要的，我们的物种保存和改变其功能， 疾病单细胞和空间技术的发展照亮了以前未知的亚型 细胞及其在子宫内膜中的空间排列，提供了有价值的见解， 参与不同子宫内膜隔室的途径，并暗示它们在决定细胞增殖中的作用。 在管腔或基底区的规格。虽然可以从这些静态图片中推断出 生理功能，从胚泡植入和侵入到大量月经出血， 捕获子宫内膜动态空间相互作用的易处理的体外模型， 关于子宫内膜功能的研究仍然具有挑战性。在这里，我们开发了子宫内膜的体外模型， 通过将组织水平的空间映射方法与体外组织工程和微流体相结合， 接近。我们将改进现有的方法，开发/测试新的模型，优先考虑鲁棒性， 可重复性，并允许更广泛的社区传播和实施，如下 目标1：设计一个强大的和可扩展的微生理系统（MPS）模型，复制空间 人子宫内膜早期至中期分泌期的小生境。我们将使用上皮细胞的共培养 以及微流体装置中的基质，其中类器官在特殊的水凝胶中经历形态发生。我们将 通过查询体内细胞图谱来告知培养基组成和其它度量。 目标2：量化细胞起源（“自然”）和空间环境（“养育”）之间的相互作用， 目的1中建立的体外模型中的上皮细胞身份。我们将使用单细胞和空间转录组学 我们将使用克隆追踪技术来绘制细胞类型的起源。通过整合 体内/体外数据集，我们将对表征 细胞源（如管腔/基底），以及扩散信号的梯度大小 分子和营养物质。 目的3：研究将内皮细胞引入体外模型的功能后果， 目标1，使用简单的“单层”协议。我们将重点关注细胞状态转换， 蜕膜化过程中内皮-间质-上皮细胞相互作用，并利用该模型研究免疫细胞 贩卖人口我们的最终目标是获得组织的系统生物学视图，这将使我们能够告知组织的结构。 开发新的治疗方法，治疗与尿道有关的疾病。 我们的工具和知识有可能使子宫内膜的临床前模型更牢固地进入临床。 人性化研究的领域这项工作对学术界和工业界都具有重要意义 研究，导致个性化的妇科医学方法。

The endometrium -- the innermost mucosal lining of the uterus that provides the site of embryo implantation- is crucial for our species preservation and alterations on its function underlay infertility and disease. Developments in single-cell and spatial technologies have illuminated previously unknown subtypes of cells and their spatial arrangements in the endometrium, providing valuable insights into the signalling pathways involved in the different endometrial compartments and hinting at their roles in determining cell specification in the luminal or basal zones. While inferences can be made from these static pictures about physiological functions ranging from blastocyst implantation and invasion to heavy menstrual bleeding, without tractable in vitro models that capture the endometrium's dynamic spatial interactions, mechanistic hypotheses about endometrial function remain challenging to test. Here, we develop in vitro models of the endometrium by combining tissue-level spatial mapping approaches with in vitro tissue engineering and microfluidic approaches. We will refine existing approaches and develop/test new models, prioritizing robustness and reproducibility, and allowing dissemination and implementation by the broader community, as follows Aim 1: Design a robust and scalable microphysiological systems (MPS) model that replicates the spatial niches of the human endometrium in the early-to-mid secretory phase. We we will use a co-culture of epithelia and stroma in a microfluidic device, in which organoids undergo morphogenesis in a special hydrogel. We will inform the media composition and other metrics by querying the in vivo cellular atlases. Aim 2: Quantify the interplay between cell origin (“nature”) and spatial environment (“nurture”) in defining epithelial cell identity in the in vitro models built in Aim 1. We will use single-cell and spatial transcriptomics technologies to profile the devices, and we will use clonal tracing to map the origin of a cell type. By integrating vivo/in vitro datasets, we will conduct a systematic investigation into the various parameters that characterize the cell source (such as luminal / basalis), as well as the magnitudes of gradients in diffusible signaling molecules and nutrients. Aim 3: Investigate the functional consequences of introducing endothelial cells into the in vitro model from Aim 1, using a simple “monolayer” protocol. We will focus on the cell state transitions that are modulated by endothelial-stromal-epithelial crosstalk during decidualization, and use the model to investigate immune cell trafficking. Our ultimate goal is to obtain a systems biology view of the tissue, which will allow us to inform the development of new treatments for endometrial-related disorders. Our tools and knowledge have potential to bring preclinical models of the endometrium more firmly into the realm of humanised research. This work has significant implications for both academic and industrial research, leading towards a personalised medicine approach for gynaecology.