Core C Cell and Tissue Imaging and Analysis

核心 C 细胞和组织成像与分析

• 批准号: 10494657
• 负责人: GEORGE F MURPHY
• 金额: $ 25.63万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-30 至 2027-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10494657
• 关键词: Age; Antibodies; Archives; Behavior; Bioinformatics; Biological Markers; Cells; Chronology; Clinical; Data; Data Analyses; Dermatopathology; Diagnostic; Ensure; Epigenetic Process; Epitopes; Event; Experimental Models; Feedback; Formalin; Functional disorder; Funding; Genomics; Histology; Human; Image; Imaging technology; Immunology; In Situ; In Vitro; Infrastructure; Integumentary system; Label; Lead; Molecular; Morphology; Mus; Natural regeneration; Paraffin Embedding; Pathology; Pathway interactions; Patients; Physicians; Process; Proteomics; Protocols documentation; Recording of previous events; Research; Research Personnel; Resolution; Resources; Role; Sampling; Scanning; Scientist; Services; Skin; Skin Aging; Skin Cancer; Specimen; Standardization; Structure of thyroid parafollicular cell; Technology; Time; Tissue imaging; Tissues; Training; Treatment Efficacy; United States National Institutes of Health; Validation; age related; base; biobank; cellular imaging; clinically relevant; comorbidity; comparative; data sharing; digital; experience; experimental study; in vivo; innovation; insight; mouse model; multiplexed imaging; novel; prospective; repaired; response; skin disorder; skin regeneration; stem cell biology; stem cells; success; targeted treatment; technology development; web site; wound healing

SUMMARY This Cell and Tissue Imaging and Analysis Core seeks provides a centralized and integrated resource for in situ proteomic analysis and imaging of the cellular and antigenic bioarchitecture of skin stem cells as a function of chronologic age and in the context of age-related deficiency in wound healing. It also provides translational validation through comparative examination of experimental murine and human skin selected as matched pairs by expert dermatopathologists to ensure cross-relevance. The Core thus combines highly skilled professional dermatopathology analysis and cutting-edge multiplex imaging technologies that permit sequential assessment of how cells and molecules behave in healing wounds, and how this behavior is compromised in the setting of age-related skin stem cell depletion/dysfunction. Specific Aim 1 provides PPG investigators with conventional histology of formalin-fixed, paraffin-embedded (FFPE) biospecimens, multiplex labeling for 3-4 epitopes, image quantification, and a variety of optional specialized technologies on an as-needed basis. This aim is essential to assess the cellular constituents and efficiency of wound healing responses in the context of various experimental models proposed in Projects 1-3 and in a manner that permits comparisons across projects. It also provides key data that will interface with the Bioinformatics Core (Core B), and assists in developing strategies to enhance wound healing through understanding potential utility of targeted therapeutic approaches directed at defective stem cell-driven repair pathways. Importantly, it offers standardized and uniform expertise and experience not available through simultaneous use of multiple institutional pathology resources. Specific Aim 2 takes multiplex labeling to the level of high-resolution, highly-plexed imaging, whereby cellular and proteomic intricacies can for the first time be visually unraveled at the level of 20 or more simultaneously identifiable antibody probes. This approach leverages in-house technology involving NanoZoomer scanning technology and QuPath data analysis in order to provide state-of-the art quantitative and spatial analysis of multiple epitopes expressed in situ by cells and their microenvironments relevant to skin aging and stem cell involvement in the wound healing process. It is fortified by collaborative interactions with Dr. Peter Sorger and application of related CyCIF labeling technology, as well as the availability of GeoMx technology for digital spatial proteomic and genomic profiling. Aim 3 provides an extensive human skin biobank whereby data generated by Core analyses of murine models may be readily compared and contrasted with relevant age-matched and co-morbidity-controlled wound healing responses in the human integument, as well as accessing and prospectively accumulating a wealth of relevant biospecimens for PPG analyses. Core C thus provides an essential, centralized, and highly integrated analytical component to the PPG, offering insight into actual cellular and molecular events that occur in situ in skin where potentially critical microenvironmental relationships remain intact, thus representing a key feedback loop in assessing experimental results, clinical relevance, and therapeutic efficacy.

概括 该细胞和组织成像和分析核心旨在为原位分析提供集中和集成的资源 皮肤干细胞的细胞和抗原生物结构的蛋白质组分析和成像作为 实际年龄以及与年龄相关的伤口愈合缺陷的情况。它还提供翻译 通过对作为配对的实验鼠和人类皮肤进行比较检查进行验证 由皮肤病理学家专家进行，以确保交叉相关性。因此，核心结合了高技能的专业人士 皮肤病理学分析和尖端多重成像技术可进行连续评估 细胞和分子在愈合伤口时如何表现，以及这种行为在以下环境中如何受到损害 与年龄相关的皮肤干细胞耗竭/功能障碍。具体目标 1 为 PPG 调查人员提供常规的 福尔马林固定、石蜡包埋 (FFPE) 生物样本的组织学，3-4 个表位的多重标记，图像 量化，以及根据需要的各种可选的专业技术。这一目标至关重要 在各种实验的背景下评估伤口愈合反应的细胞成分和效率 项目 1-3 中提出的模型以及允许跨项目进行比较的方式。它还提供了密钥 数据将与生物信息学核心（核心 B）接口，并协助制定策略以增强 通过了解针对缺陷的靶向治疗方法的潜在效用来治愈伤口 干细胞驱动的修复途径。重要的是，它提供标准化和统一的专业知识和经验，而不是 通过同时使用多个机构病理学资源来获得。具体目标 2 采用多重方式 标记到高分辨率、高度复杂的成像水平，从而可以分析细胞和蛋白质组的复杂性 首次在 20 个或更多同时可识别的抗体探针的水平上以视觉方式解开。这 该方法利用了涉及 NanoZoomer 扫描技术和 QuPath 数据分析的内部技术 为了对细胞原位表达的多个表位提供最先进的定量和空间分析 及其与皮肤衰老和干细胞参与伤口愈合过程相关的微环境。它 与 Peter Sorger 博士的合作互动以及相关 CyCIF 标签的应用得到了加强 技术，以及用于数字空间蛋白质组和基因组分析的 GeoMx 技术的可用性。 Aim 3 提供了一个广泛的人类皮肤生物库，其中通过小鼠模型的核心分析生成数据 可以很容易地与相关的年龄匹配和共病控制的伤口愈合进行比较和对比 人类外皮的反应，以及获取和前瞻性地积累大量相关的 用于 PPG 分析的生物样本。因此，Core C 提供了一个必要的、集中的、高度集成的分析 PPG 的组成部分，可深入了解皮肤原位发生的实际细胞和分子事件，其中 潜在的关键微环境关系保持完整，因此代表了关键的反馈循环 评估实验结果、临床相关性和治疗效果。