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.

总结 该细胞和组织成像和分析核心旨在为原位成像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学分析和成像， 年龄和与年龄相关的伤口愈合缺陷的背景下。它还提供了翻译 通过选择作为匹配对的实验鼠和人皮肤的比较检查进行验证 以确保交叉相关性。因此，核心结合了高技能的专业人员， 皮肤病理学分析和尖端的多重成像技术，允许连续评估 细胞和分子在愈合伤口中的行为，以及这种行为如何在 与年龄相关的皮肤干细胞耗竭/功能障碍。特定目标1为PPG研究者提供了常规的 福尔马林固定、石蜡包埋（FFPE）生物标本组织学，3 - 4个表位的多重标记，成像 量化和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，并以允许跨项目进行比较的方式进行。它还提供了关键 将与生物信息学核心（核心B）接口的数据，并协助制定战略， 通过了解针对缺陷性伤口的靶向治疗方法的潜在效用， 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， 可通过同时使用多个机构病理学资源获得。Specific Aim 2采用多路复用 标记到高分辨率，高度复杂的成像水平，从而细胞和蛋白质组的复杂性可以 第一次在20个或更多个同时可识别的抗体探针的水平上被视觉解开。这 该方法利用了内部技术，包括NanoZoomer扫描技术和QuPath数据分析 为了提供细胞原位表达的多个表位的现有技术的定量和空间分析 以及它们与皮肤老化和干细胞参与伤口愈合过程相关的微环境。它 通过与Peter Sorger博士的协作互动以及相关CyCIF标签的应用得到了加强 技术，以及GeoMx技术用于数字空间蛋白质组学和基因组分析的可用性。 目的3提供了一个广泛的人类皮肤生物库，其中通过对小鼠模型的核心分析产生的数据 可以容易地与相关年龄匹配和共病对照的伤口愈合进行比较和对比 在人类外皮的反应，以及访问和前瞻性地积累了丰富的相关 用于PPG分析的生物样本。因此，核心C提供了一个基本的、集中的、高度集成的分析 PPG的组成部分，提供对皮肤中原位发生的实际细胞和分子事件的深入了解， 潜在的关键微环境关系保持不变，因此代表了一个关键的反馈回路， 评估实验结果、临床相关性和治疗功效。