Neuronal anatomy, connectivity, and phenotypic innervation of the knee joint

膝关节的神经元解剖学、连接性和表型神经支配

基本信息

批准号：
10608851
负责人：
Benjamin R Arenkiel
金额：
$ 738.87万
依托单位：
BAYLOR COLLEGE OF MEDICINE
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-09-23 至 2025-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10608851
关键词：
3-Dimensional Adenoviruses Afferent Neurons Age Anatomy Animal Model Autonomic nervous system Behavior Bioinformatics Biology Biopsy Blood Vessels Cartilage Cells Clinical Trials Complex Data Data Science Degenerative polyarthritis Development Disease Endothelial Cells Enterobacteria phage P1 Cre recombinase Exercise Fascia GTP-Binding Proteins Gender Genes Genetic Genetic Models Health Homeostasis Inflammation Infrastructure Interleukin-1 Intervention Joint Capsule Joints Knee Osteoarthritis Knee joint Knowledge Ligaments Maps Measures Medial meniscus structure Mediating Methodology Methods Modeling Molecular Molecular Profiling Morbidity - disease rate Mus Muscle Neuroanatomy Neurons Operative Surgical Procedures Opiate Addiction Pain Pattern Phenotype Physical activity Play Procedures Quality of life Rabies Rabies virus Receptor Activation Reporter Reporting Resolution Role Running Science Sensory Specificity Spinal Ganglia Surgical Models Techniques Technology Tendon structure Therapeutic Time Tissue imaging Tissues Translations Validation Vascularization Viral Viral Vector Virus Visualization age effect animal tissue arthropathies bone cell type combinatorial data management gene therapy high dimensionality human tissue improved molecular phenotype mortality mouse Cre recombinase neovascularization nerve supply neural circuit neuronal circuitry neuronal patterning new technology new therapeutic target opioid use pain perception pre-clinical protein biomarkers response response to injury retrograde transport sex single-cell RNA sequencing skeletal targeted treatment therapeutic target three-dimensional visualization tool transcriptomics translational impact translational potential two-dimensional

项目摘要

PROJECT SUMMARY Identifying patterns of neuronal connectivity is critical for understanding functional and anatomical circuits that mediate pain perception. However, knowledge about the types and distribution of neurons in joint tissues have generally been limited to traditional 2-dimension histopathological and immunohistopathological approaches, and little to no information is available on connectivity and neuronal phenotypes. New technologies have emerged that allow for both trans-synaptic circuit analysis and precise control of neuronal firing, including the use of retrogradely transported viral vectors (i.e., pseudotyped rabies virus) and heterologous receptor activation. At the same time, 3-dimensional visualization of neuronal and vascular patterns have been advanced by tissue clearing techniques in conjunction with cell type specific fluorescent markers generated by intercrossing cell type specific Cre recombinase mouse lines with a variety of conditionally activated reporters. Finally, the advent of single cell RNA sequencing has allowed for extending cellular phenotyping to a molecular level that has not only increases analytic resolution, but also therapeutic targeting with greater disease specificity than previously possible. The development of high resolution spatial transcriptomics, i.e., MERFISH, allows for correlation and validation of scRNA-seq data. In this context, osteoarthritis of the knee joint is an optimal model for applying these tools as abundant genetic and surgical models are available for orthogonal validation of findings. Moreover, in the preclinical context, various therapeutic approaches including gene therapy have been shown to impact pain measures, and as such, they constitute an important interventional validation of molecular changes that are identified in neurons in the disease state. The fact that some of these therapies are now in clinical trial adds to the potential translational impact of the proposed preclinical findings here. Ultimately, the combination of both anatomic, 3-D, and molecular signatures will facilitate the translation into human tissues and biopsies, while maximizing the likelihood of relevant new therapeutic targets.

项目概要识别神经元连接模式对于理解功能和解剖回路至关重要介导疼痛感知。然而，关于关节组织中神经元的类型和分布的知识已经通常仅限于传统的二维组织病理学和免疫组织病理学方法，关于连通性和神经元表型的信息很少甚至没有。新技术有出现了允许跨突触回路分析和神经元放电精确控制的技术，包括使用逆行运输的病毒载体（即假型狂犬病病毒）和异源受体激活。同时，神经元和血管模式的 3 维可视化已被通过组织透明技术结合细胞类型特异性荧光标记物产生的先进技术细胞类型特异性 Cre 重组酶小鼠系与多种条件激活报告基因的交叉。最后，单细胞 RNA 测序的出现使得细胞表型分析扩展到分子水平。不仅提高了分析分辨率，而且还针对更大的疾病进行了治疗靶向比以前可能的特异性。高分辨率空间转录组学的发展，即MERFISH，允许 scRNA-seq 数据的关联和验证。在这种情况下，膝关节骨关节炎是一种应用这些工具的最佳模型，因为丰富的遗传和手术模型可用于正交验证调查结果。此外，在临床前背景下，包括基因治疗在内的各种治疗方法治疗已被证明可以影响疼痛测量，因此，它们构成了重要的介入治疗验证疾病状态下神经元中发现的分子变化。事实上，其中一些目前正在进行临床试验的疗法增加了所提出的临床前研究结果的潜在转化影响这里。最终，解剖学、3D 和分子特征的结合将促进翻译进入人体组织和活检，同时最大限度地提高相关新治疗靶点的可能性。