In vivo label free optical imaging of immune cells in human skin

人体皮肤免疫细胞体内无标记光学成像

• 批准号: 10664746
• 负责人: Mihaela Balu
• 金额: $ 17.27万
• 依托单位: UNIVERSITY OF CALIFORNIA-IRVINE
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-03-01 至 2024-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10664746
• 关键词: Address; Animal Experiments; Animal Model; Antibodies; Architecture; Area; Atopic Dermatitis; Autoimmune; Autoimmune Diseases; Autoimmune Process; Basal cell carcinoma; Binding Proteins; Biological Assay; Biological Markers; Blood Vessels; Cell Communication; Cells; Cellular Metabolic Process; Cellular Morphology; Chemicals; Clinical; Clinical Research; Collaborations; Cutaneous; Dermal; Detection; Devices; Disease; Disease remission; Early Diagnosis; Environment; Fluorescence; Germany; Goals; Heterogeneity; Histologic; Human; Image; Imaging Device; Immune; Immune response; Immune system; Immunologic Monitoring; Immunotherapy; Inflammatory; Investigation; Knowledge; Label; Lesion; Malignant Neoplasms; Measurement; Measures; Melanins; Metabolic; Methods; Microscopic; Modeling; Molecular; Morphology; Mus; NADH; Optics; Pathogenesis; Performance; Phase; Pigments; Play; Population; RNA Probes; Remission Induction Therapy; Reporter; Reporting; Resolution; Role; Scanning; Signal Transduction; Skin; Skin Cancer; Skin Carcinoma; Spatial Distribution; Specificity; Speed; Structure; Techniques; Technology; Three-Dimensional Image; Time; Tissues; Validation; Visualization; Work; cancer therapy; cell type; clinical imaging; combinatorial; density; design; detection sensitivity; fluorophore; human imaging; imaging approach; imaging platform; immune activation; immune imaging; immunoreaction; improved; in vivo; in vivo imaging; microscopic imaging; mouse model; multi-photon; multiphoton microscopy; optical imaging; response; skin disorder; spatiotemporal; success; tumor-immune system interactions

Summary. The study of mammalian immune cells and their interactions under native conditions is critical for understanding the mechanisms initiating many diseases, including cancer, and for improving their treatments. Intravital multiphoton microscopy (MPM), fluorescent reporter mouse models and in vivo cell and tissue labeling techniques have made possible the investigation of immune cells interactions at a cellular/subcellular level in their native environment. However, the significant differences in the skin and the immune system of mice and humans preclude studies of uniquely human immune responses. The goal of this proposal is to evaluate the ability of intravital MPM imaging to identify and distinguish immune cell populations in human skin, based on label-free molecular contrast. We will utilize a fast, large area multiphoton exoscope (FLAME), an imaging platform recently designed and developed by our group to have unique performance features optimized for clinical skin imaging. FLAME has the ability to generate in vivo 3D images of human skin over macroscopic areas (up to 8x8 mm2) with microscopic resolution (0.5-1µm) at fast acquisition rates (tens of seconds). FLAME features label-free specificity for melanin and NADH based on their spectral and rapid time-resolved endogenous fluorescence detection. Selective detection of melanin allows for imaging of pigment-rich cells such as melanophages with high specificity, while time-resolved NADH fluorescence detection reports on the protein binding activity of these molecules within the immune cells and on their metabolic heterogeneity. We have demonstrated the ability of FLAME to detect resident immune cells in normal human skin as well as an activation of a cellular immune response, based on endogenous molecular contrast. In this proposal, we will develop methods for the in vivo characterization of the spatial and metabolic heterogeneity of immune cells in human skin by using atopic dermatitis as an inflammatory skin condition model and Multi Omic Single-scan Assay with Integrated Combinatorial Analysis (MOSAICA) for validation. We will also evaluate the feasibility of FLAME to quantify changes in immune cell metabolism and spatiotemporal heterogeneity in response to immune activation. If successfully validated in a larger clinical study, this approach would be a critical first step in making it possible to characterize cellular-level immune responses in human skin at the bedside, with broad applications ranging from detecting early immune reactions to developing improved cancer treatments.

概括。 对哺乳动物免疫小球及其在天然条件下的相互作用的研究对于理解至关重要 引发许多疾病的机制，包括癌症，以及改善其治疗方法。弹药 多光子显微镜（MPM），荧光记者小鼠模型以及体内细胞和组织标记 技术使Immunocells相互作用在细胞/亚细胞水平上的投资成为可能 他们的本地环境。但是，小鼠皮肤和免疫系统的显着差异以及 人类排除了独特的人免疫复杂的研究。该提议的目的是评估 浸润室MPM成像识别和区分人皮肤中免疫群人群的能力 无标签的分子对比。我们将利用快速的大面积多光外观（火焰），成像 我们小组最近设计和开发的平台旨在具有优化的独特性能功能 临床皮肤成像。火焰有能力在宏观区域产生人体皮肤的体内3D图像 （高达8x8 mm2）以快速采集率（数十秒）以显微镜分辨率（0.5-1μm）。火焰功能 基于它们的光谱和快速时间分辨的内生的黑色素和NADH的无标签特异性 荧光检测。黑色素的选择性检测允许成像富含色素的细胞，例如 具有高特异性的素噬细胞，而时间分辨的NADH荧光检测报告有关蛋白质 这些分子在免疫细胞内及其代谢异质性的结合活性。我们有 证明了火焰检测正常皮肤中居民免疫细胞的能力以及激活 基于内源分子对比的细胞免疫反应。在此提案中，我们将发展 人体内表征人体内的体内表征的方法和代谢异质性 通过将特应性皮炎用作炎症皮肤状况模型和多余的单胶质测定 综合组合分析（Mosaica）进行验证。我们还将评估火焰的可行性 响应免疫激活而量化免疫球代谢和空间时间异质性的变化。 如果在较大的临床研究中成功验证，这种方法将是使它成为可能的关键第一步 为了表征床边人皮肤中的细胞水平免疫反应，范围广泛 从检测早期免疫反应到发展改善的癌症治疗方法。