Establishing an immune mechanome

摘要 作为炎症反应的结果，机械力经常在病变或受伤的组织中被调节 由受影响部位的免疫细胞活动驱动(S)。事实上，病理性的异常机械力的产生 环境可能调节疾病进展和治疗耐药性。然而，人们对这种反应知之甚少。 免疫细胞对这些机械力的反应，特别是在组织长度的尺度上，甚至在正常情况下 生理环境。因此，有一种严重未得到满足的需要，即填补我们对 组织水平的机械力和免疫细胞行为之间的相互作用，无论是集体还是在 单细胞水平。在NIGMS R35 Mira的支持下，在未来五年内进行早期调查， 我的实验室将建立第一个免疫机制。我们将研究组织机械的影响 对各种器官中的先天免疫细胞和适应性免疫细胞的表型和功能的影响。利用 基于工程的工具和方法，我们将在以下位置将无偏见组学平台与机械测试相结合 多个尺度(例如，体内细胞、体外组织和体内器官)，以便与免疫反应相关 机械力。在多尺度压缩期间，贩运、分布、运动性、细胞-细胞相互作用以及 免疫细胞的功能行为将通过以下方式进行检查和干扰：i)活体和动态成像(例如， 利用荧光细胞或基因工程小鼠模型的多光子显微镜)；ii)具有免疫活性， 转基因和免疫动物模型(例如，OT-I/OT-II抗原系统)；以及iii)人工智能- 基于细胞状态分析(例如，来自单细胞RNA测序)。我们还将探讨我们的假设 面对病理情况时的有益免疫活动被增强的组织机械作用所抑制 力量。重要的是，拟议的项目将在独立于 组织类型、器官或疾病，以便最大限度地发挥在生物医学科学中广泛影响的潜力。 所产生的知识将为未来在这两个领域的机械学和翻译研究奠定基础 健康和患病的环境。通过在机械工程和免疫学的界面上操作 在“机械免疫学”这一新兴领域，我的研究项目非常适合揭示新的生物物理学。 人类疾病的洞察力和病理生理学目标。

ABSTRACT Mechanical forces are often modulated in diseased or wounded tissues as a result of inflammatory responses driven by immune cell activity in the affected site(s). In fact, aberrant mechanical force generation in pathological settings may mediate disease progression and treatment resistance. However, little is known about the response of immune cells to these mechanical forces, particularly at the tissue-length scale, and even in normal physiological settings. Thus there is a critically unmet need to fill overlooked gaps in our basic understanding of the interplay between tissue-level mechanical forces and immune cell behavior, both collectively and at the single-cell level. With the support of the NIGMS R35 MIRA for Early Stage Investigators over the next five years, my laboratory will establish the first immune mechanome. We will investigate the impact of tissue mechanical forces on the phenotype and function of innate and adaptive immune cells in a variety of organs. Leveraging engineering-based tools and approaches, we will couple unbiased omics platforms to mechanical testing at multiple scales (e.g., on cells in vivo, tissues ex vivo, and organs in vivo) in order to relate immune response to mechanical forces. During multiscale compression, the trafficking, distribution, motility, cell-cell interactions, and functional behavior of immune cells will be examined and perturbed via: i) intravital and dynamic imaging (e.g., with multiphoton microscopy of fluorescent cells or genetically engineered mouse models); ii) immunocompetent, transgenic, and immunogenic animal models (e.g., OT-I/OT-II antigen systems); and iii) artificial intelligence- based cell state analysis (e.g., from single cell RNA sequencing). We will also explore our hypothesis that beneficial immune activity in the face of pathological conditions is suppressed by heightened tissue mechanical forces. Importantly, the proposed Projects are to be performed in non-specific contexts that are independent of tissue type, organ, or disease in order to maximize the potential for broad impact in the biomedical sciences. The knowledge generated will lay the groundwork for future mechanistic and translational research in both healthy and diseased settings. By operating at the interface of mechanical engineering and immunology in the burgeoning field of “mechano-immunology,” my research program is uniquely suited to reveal new biophysical insights and pathophysiological targets for human disease.