The immune system beyond protection: role of innate lymphoid cells in the regulation of the estrous cycle.

免疫系统无法保护：先天淋巴细胞在动情周期调节中的作用。

• 批准号: RGPIN-2021-03885
• 负责人: Ardolino, Michele
• 金额: $ 2.19万
• 依托单位: University of Ottawa
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2022
• 资助国家: 加拿大
• 起止时间: 2022-01-01 至 2023-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=744310
• 关键词: immune; system; beyond; protection; role

Perturbations to steady state conditions are sensed by tissue resident immune cells, which elaborate the information received and respond by producing signals that shape the functions of other immune, as well as non-immune, cells. In recent years, it became clear that such perturbations are not only caused by harmful pathogens, rather immune cells play fundamental roles in many physiological processes not related to host protection. Major players in these scenarios are Innate Lymphoid Cells (ILCs) and myeloid cells, which populate the connective tissues underneath the epithelial layer of barrier organs and prime inflammatory responses through the production of different cytokines. Such signals then lead to the recruitment and activation of circulating lymphocytes, which mediate a response. In this research program, we propose to study immune cells in the female reproductive tract, a dynamic system which undergoes dramatic changes during the estrous cycle. Our working hypothesis is that immune cells are important regulators of the dynamic changes occurring during the estrous cycle. We will carry this project in collaboration with Dr. Vanderhyden, an expert in reproductive biology. In line with our research program, the short-term objectives of this Discovery Grant are: 1)To build an atlas of innate lymphoid cells in the female reproductive system: We will use single-cell RNA-sequencing (scRNA-seq) to characterize the immune composition of the female reproductive system. Ovaries, oviducts and uteri will be analyzed from mice at various stages of the estrous cycle and the periovulatory period. This analysis will reveal which immune cell populations and which pathways are modulated at the different stages of the estrous cycle. Immunohistochemistry, qPCR and flow cytometry will be employed to corroborate the scRNA-seq results. A particular emphasis will be given to innate lymphoid cells, which have been vastly overlooked in the female reproductive tract. 2)To determine if and how innate lymphoid cells regulate the progression through the estrous cycle: As we hypothesize that innate lymphoid cells are important coordinators of the dynamics of the estrous cycle, we will determine how experimental perturbation of this immunological compartment affects the estrous cycle. 3)To understand how endocrine signals regulate innate lymphoid cell functions during the estrous cycle: As innate lymphoid cells express receptors for neurotransmitters and hormones, we will study if they are directly regulated by the neuro-endocrine system. We will conduct functional in vivo and ex vivo experiments to determine which neuro endocrine signals regulate immune functions. This project will shed light on how immune cells regulate the estrous cycle and will represent a stepping stone to our broader goal of determining how the immune system contributes to the host physiology in the absence of a pathogenic challenge.

稳态条件的扰动由组织驻留免疫细胞感测，其通过产生塑造其他免疫细胞以及非免疫细胞的功能的信号来阐述所接收的信息并作出响应。近年来，人们越来越清楚，这种扰动不仅是由有害的病原体引起的，而且免疫细胞在许多与宿主保护无关的生理过程中发挥着重要作用。这些情况中的主要参与者是先天性类淋巴细胞（ILC）和髓样细胞，其填充屏障器官的上皮层下方的结缔组织，并通过产生不同的细胞因子引发炎症反应。然后，这些信号导致循环淋巴细胞的募集和活化，其介导应答。在这项研究计划中，我们建议研究女性生殖道中的免疫细胞，这是一个动态系统，在发情周期中会发生巨大变化。我们的工作假设是，免疫细胞是发情周期中发生的动态变化的重要调节器。我们将与生殖生物学专家Vanderhyden博士合作开展这个项目。 根据我们的研究计划，这项发现资助的短期目标是：1）建立女性生殖系统中先天淋巴细胞的图谱：我们将使用单细胞RNA测序（scRNA-seq）来表征女性生殖系统的免疫组成。将在发情周期和排卵期的不同阶段分析小鼠的卵巢、输卵管和子宫。这种分析将揭示哪些免疫细胞群和哪些途径在发情周期的不同阶段受到调节。将采用免疫组织化学、qPCR和流式细胞术来证实scRNA-seq结果。一个特别的重点将给予先天淋巴细胞，这已被大大忽视了在女性生殖道。2）确定先天淋巴细胞是否以及如何调节发情周期的进程：由于我们假设先天淋巴细胞是发情周期动力学的重要协调者，我们将确定这种免疫区室的实验扰动如何影响发情周期。3）了解内分泌信号如何调节先天淋巴细胞在发情周期中的功能：由于先天淋巴细胞表达神经递质和激素的受体，我们将研究它们是否直接受神经内分泌系统的调节。我们将进行功能性的体内和体外实验，以确定哪些神经内分泌信号调节免疫功能。该项目将阐明免疫细胞如何调节发情周期，并将代表我们更广泛的目标，即确定免疫系统如何在没有病原体挑战的情况下对宿主生理学做出贡献的垫脚石。