Validation of Prenatal Rabbit Hypoxia Ischemia as a Model of Cerebral Palsy-induced Pain

产前兔缺氧缺血作为脑瘫引起的疼痛模型的验证

• 批准号: 10813313
• 负责人: MEGAN R DETLOFF
• 金额: $ 272.7万
• 依托单位: UNIVERSITY OF RHODE ISLAND
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-19 至 2026-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10813313
• 关键词: ARHGEF5 gene; Address; Affect; Age; Aging; Anatomy; Animal Model; Anxiety; Area; Arthritis; Behavior; Behavior assessment; Birth; Brain; Brain Hypoxia-Ischemia; Brain Injuries; CDK6-associated protein p18; Cerebral Palsy; Chronic; Classification; Clinical; Cognition; Conflict (Psychology); Contracture; Data; Databases; Development; Etiology; Event; Exhibits; Experimental Models; Face; Family; Fiber; Functional disorder; Hand; Hip region structure; Histology; Hyperalgesia; Hyperreflexia; Hypersensitivity; Hypoxia; Impairment; Individual; Infection; Inflammation; Injury; Ischemia; Joints; Knowledge; Leporidae; Link; Lower Extremity; Measurement; Measures; Mechanics; Medical; Mental Depression; Modeling; Morbidity - disease rate; Motor; Movement; Multivariate Analysis; Muscle; Muscle Hypertonia; Muscle Spasticity; Musculoskeletal; Musculoskeletal Pain; Nature; Nerve Fibers; Neurons; Neuropathy; New Zealand; Nociception; Nociceptors; Oryctolagus cuniculus; Outcome; Pain; Pain Measurement; Pain management; Pathway interactions; Patient Self-Report; Perinatal; Perinatal Hypoxia; Peripheral; Persons; Pharmaceutical Preparations; Phenotype; Physiological; Placental Insufficiency; Pregnancy; Procedures; Psychological Stress; Quality of life; Reporting; Reproducibility; Research; Research Personnel; Rodent; Rodent Model; Schizophrenia; Sensory; Severities; Site; Spastic Cerebral Palsy; Spinal Cord; Stress; Sucrose; Surgeon; Syndrome; System; Testing; Torque; Training; Validation; Validity and Reliability; anxiety-like behavior; autism spectrum disorder; biopsychosocial; chronic pain; chronic pain patient; comorbidity; comparative; density; depressive symptoms; detection sensitivity; disability; experience; experimental group; foot; gabapentin; motor deficit; motor disorder; motor impairment; neonate; neural; novel; object recognition; pain perception; pain relief; pain sensitivity; painful neuropathy; patch clamp; physical insult; postnatal; preclinical study; preference; prenatal; prescription opioid; sex; spastic paralysis; subluxation; therapeutic target; validation studies; virtual; white matter injury

Project Summary / Abstract Cerebral palsy (CP) is a condition defined by motor impairment, but pain is the most prevalent co-morbidity. Pain is commonly attributed to stiff, spastic muscles, joint contractures, hip subluxation and arthritis. However, the presence of chronic pain does not neatly correlate with severity of musculoskeletal problems and may be entirely independent (ie thermal hypersensitivity). CP associated pain can be both nociplastic and neuropathic in nature and could be caused by the same global neural damage that disrupted movement. Research on pain in CP requires using an animal model that displays both phenotypes of motor dysfunction and enhanced pain. We address this clinical need by validating a larger, non-rodent animal model of CP which exhibits both sensory and motor impairments after prenatal hypoxia. New Zealand White (NZW) rabbits (family Leporidae) have higher face, construct and criterion validity than rodent models of CP and are widely used for comparative preclinical studies of CP. Recent data from our labs reveal these rabbits also have enhanced nociception. We propose to further validate HI rabbits as a model of chronic pain in CP, an area that is completely unstudied. Aim 1 Determine face validity of HI rabbits as a model of CP-associated pain. Anatomical and physiological evidence of musculoskeletal pain, nociplastic pain and neuropathic pain will be assessed in naïve control, HI affected, and sham-operated rabbit kits from birth to sexual maturity (at P5, P18, P60, P180). We will characterize nociceptor dysfunction using patch clamp of DRG neurons (n = 100 neurons from 10 rabbits per age per experimental group), histology of peripheral nociceptors (sprouting of central afferent fibers, counts of nociceptors in the DRGs, and epidermal nerve fiber density in 6 kits per age per experimental group), and musculoskeletal impairment (modified Ashworth, joint torque and muscle shortening in all kits in Aims 1-2). Aim 2 Validate that sensory behavior in HI rabbits is consistent with pain phenotypes in CP. We will build upon our initial studies on nociception in neonates to verify the progression of behaviors related to biopsychosocial pain during aging. Specifically, we will perform tests of nociception, pain perception, cognition, and depressive- and anxiety-like behaviors in 20 kits per sex per experimental group, measured longitudinally as in Aim 1. A multivariate analysis will compare measures in control, sham and HI affected groups, sex and severity of motor deficits. All tests will be assessed for reliability and validity. The most efficient and valid procedures will be identified to maintain high sensitivity for detection of pain. These tests will determine if HI rabbits show robust, reproducible behaviors indicative of the multifaceted pain similar to people with CP. Aim 3 Determine reproducibility and reliability of the rabbit HI model of CP for studies of pain. Model reproducibility will be determined by having experienced surgeons, caretakers, and behaviorists train novice investigators. Inter-experimenter reliability will be assessed in all procedures and assessments in Aims 1 - 2. Together, these tests will provide a validation of the HI rabbit for use in studies of pain in the context of CP.

项目概要/摘要 脑瘫 (CP) 是一种以运动障碍为特征的疾病，但疼痛是最常见的合并症。 疼痛通常归因于肌肉僵硬、痉挛、关节挛缩、髋关节半脱位和关节炎。 慢性疼痛的存在与肌肉骨骼问题的严重程度并不完全相关，并且可能与 完全独立的（即热过敏）相关的疼痛可以是伤害性的和神经性的。 在自然界中，可能是由扰乱疼痛研究的相同的全球神经损伤引起的。 脑瘫的治疗需要使用同时表现出运动功能障碍和疼痛加剧的表型的动物模型。 我们通过验证更大的非啮齿动物 CP 动物模型来满足这一临床需求，该模型表现出 产前缺氧后的感觉和运动障碍。新西兰白兔（兔科）。 与CP啮齿动物模型相比，具有更高的面值、结构和标准效度，广泛用于比较 我们实验室的最新数据表明，这些兔子的伤害感受也有所增强。 建议进一步验证 HI 兔子作为 CP 慢性疼痛的模型，这是一个完全未经研究的领域。 目标 1 确定 HI 兔子作为 CP 相关疼痛模型的表面有效性。 肌肉骨骼疼痛、伤害性疼痛和神经性疼痛的生理学证据将在幼稚时进行评估 对照组、HI 受影响组和假手术组兔从出生到性成熟（P5、P18、P60、P180）。 将使用 DRG 神经元的膜片钳来表征伤害感受器功能障碍（n = 10 个来自 10 只兔子的 100 个神经元） 每个实验组的年龄），外周伤害感受器的组织学（中枢传入纤维的萌发，计数 DRG 中伤害感受器的数量，以及每个实验组每个年龄 6 个试剂盒中的表皮神经纤维密度），以及 肌肉骨骼损伤（目标 1-2 中所有套件中的改良 Ashworth、关节扭矩和肌肉缩短）。 目标 2 验证 HI 兔子的感觉行为与 CP 的疼痛表型一致。 以我们对新生儿伤害感受的初步研究为基础，以验证与以下行为相关的行为进展 具体来说，我们将进行伤害感受、疼痛感知、认知、 纵向测量每个实验组每个性别 20 个实验组的抑郁和焦虑样行为 如目标 1 所示。多变量分析将比较对照组、假手术组和 HI 受影响组的测量值、性别和 所有测试都将进行可靠性和有效性评估。 将确定程序以保持疼痛检测的高灵敏度。这些测试将确定是否存在 HI。 兔子表现出稳健、可重复的行为，表明与 Aim 3 患者相似的多方面疼痛。 确定用于疼痛模型研究的兔 HI 模型的再现性和可靠性。 可重复性将取决于经验丰富的外科医生、护理人员和行为学家对新手的培训 研究人员之间的可靠性将在目标 1 - 2 的所有程序和评估中进行评估。 总之，这些测试将验证 HI 兔子用于 CP 背景下的疼痛研究。