Non-viral Reprogramming of Intervertebral Disc Cells for the treatment of Discogenic back pain

椎间盘细胞非病毒重编程治疗椎间盘源性背痛

• 批准号: 9911144
• 负责人: Natalia Higuita-Castro
• 金额: $ 36.77万
• 依托单位: OHIO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-20 至 2022-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9911144
• 关键词: Acute; Address; Adolescence; Adult; Animal Model; Automobile Driving; Behavior; Biological; Biomedical Engineering; Blood Vessels; Brachyury protein; Canis familiaris; Catabolism; Cell Culture Techniques; Cells; Cellularity; Chronic; Chronic low back pain; Clinical; Clinical Trials; Cognition; Communities; Control Groups; DNA; Data; Development; Disease; Dog Diseases; Drug Addiction; Economic Burden; Endothelial Cells; Engineering; Extracellular Matrix; FOXF1 gene; Failure; Fibroblasts; Fibrosis; Functional disorder; GAG Gene; Gene Proteins; Generations; Genetic; Genome; Glycosaminoglycans; Goals; Health; Height; Human; Hyperalgesia; In Situ; In Vitro; Inflammation; Inflammatory; Injury; Intervertebral disc structure; Lead; Low Back Pain; Magnetic Resonance Imaging; Mechanics; Medical; Methods; Modeling; Mus; Nerve; Operative Surgical Procedures; Opioid; Pain; Pathologic; Pathology; Patients; Phase; Phase Transition; Phenotype; Public Health; Puncture procedure; Quality of life; Research; Research Personnel; Research Priority; Rodent Model; Societies; Somatic Cell; Speed; Stem cells; Structure; System; Technology; Therapeutic; Therapeutic Effect; Tissues; Transfection; Translating; Translational Research; Treatment Efficacy; United States National Institutes of Health; Validation; Vertebral column; Veterinarians; Viral Vector; Work; addiction; clinically relevant; cytotoxicity; discogenic pain; extracellular; extracellular vesicles; gene therapy; healing; human disease; improved; in vivo; in vivo Model; inflammatory pain; innovation; interdisciplinary approach; intervertebral disk degeneration; minimally invasive; mouse model; multidisciplinary; nano; neurotrophic factor; novel; nucleus pulposus; opioid epidemic; optimal treatments; pain behavior; pain model; pain patient; pain relief; pain symptom; pre-clinical; programs; protein expression; reduce symptoms; restoration; socioeconomics; spontaneous pain; stem; stem cell therapy; tool; transcription factor; translational approach; translational model

Project Summary/Abstract Chronic low back pain exerts a significant socio-economic burden on society and is a major contributor to the growing opioid crisis. Last year, NIH “launched the HEAL (Helping to End Addiction Long-term) initiative, an aggressive, trans-agency effort to speed scientific solutions to stem the national opioid public health crisis”. This enormous burden is largely because studies have failed to target the underlying mechanisms associated with pain generation. Intervertebral disc (IVD) degeneration is strongly associated with the pathophysiology of low back pain and identifying non-addictive minimally invasive treatments for discogenic back pain (DBP) is a research priority. Pathological IVD changes include extracellular matrix (ECM) breakdown, inflammation and aberrant nerve/vascular ingrowth which have been shown to significantly correlate with pain. Therefore the optimal therapy for DBP would target both structural restoration and reduce the symptoms of pain. Yet current strategies involving the use of stem cells or gene therapy are faced with a number of challenges which include failure of stem cells to adapt to the harsh IVD microenvironment, the use of viral vectors and unwarranted DNA deletions within the host genome. Furthermore clinicians do not have access to clinically relevant tools or technologies that could directly help treat the underlying disease in patients with DBP. There is a critical need for a biological non-addictive strategy that addresses these limitations. Our goal is to use novel cellular reprogramming technologies to alter the innate cell phenotype of native diseased IVD cells to a healthy extracellular producing and anti-catabolic/inflammatory phenotype in human in vitro cell culture and in vivo models of DBP. Non-viral delivery for transporting genetic cargo into the cell such as engineered extracellular vesicles (EVs) or tissue nano-transfection (TNT) offer safe and minimally invasive methods for reprogramming somatic cells and recent work by the investigators has demonstrated successful reprogramming of adult fibroblasts into endothelial cells in vivo mouse models. We propose using these innovative non-viral delivery systems to deliver genetic cargo to IVDs in vitro and in vivo. The first specific aim (R61 Phase 1 Aim 1) focuses on the effects of EV or TNT delivery of transcription factors on diseased human nucleus pulposus cells and tissue in vitro examining changes in ECM and catabolic, inflammatory and pain markers. The second aim (R61 Phase 1 Aim 2) investigates the effects of EV or TNT delivery of transcription factors in mouse IVD puncture models of DBP assessing changes in disc structure/function, pain, cognition and cytotoxicity. These studies are both significant and highly innovative because they combine a unique multi-disciplinary team of medical and veterinary clinicians, spine biologists, neuroscientists, biomedical engineers, and a biostatistician to interrogate the use of these novel concepts and technologies to treat DBP. The broader impacts of this proposal and transition to the R33 portion involve assessing this strategy in clinically relevant chondrodystrophic dogs that develop DBP spontaneously followed by clinical trials in patient dogs with DBP.

项目摘要/摘要 慢性下腰痛给社会带来了巨大的社会经济负担，是导致 日益严重的阿片类药物危机。去年，美国国立卫生研究院发起了Hear(帮助结束长期上瘾)倡议， 积极、跨机构的努力，以加快科学解决方案，以遏制国家阿片类药物公共卫生危机“。 这种巨大的负担在很大程度上是因为研究未能针对相关的潜在机制 伴随着痛苦的产生。椎间盘退变与腰椎间盘突出症的病理生理密切相关。 下腰痛和识别非成瘾性微创疗法治疗间盘源性腰痛(DBP)是一种 研究优先。病理性IVD改变包括细胞外基质(ECM)破坏、炎症和 异常的神经/血管内生，已被证明与疼痛显著相关。因此， DBP的最佳治疗将以结构恢复和减轻疼痛症状为目标。目前还很流行 涉及使用干细胞或基因疗法的战略面临一些挑战，包括 干细胞无法适应恶劣的IVD微环境、使用病毒载体和不必要的DNA 宿主基因组中的缺失。此外，临床医生无法获得临床相关工具或 可以直接帮助治疗DBP患者潜在疾病的技术。有一种迫切的需要 寻求一种生物上的非成瘾策略来解决这些限制。我们的目标是使用新的蜂窝 重新编程技术将自然疾病IVD细胞的固有细胞表型改变为健康的 人体外细胞培养和体内细胞外产生及抗分解/炎症表型 DBP的模型。非病毒递送，用于将基因货物运送到细胞内，如工程细胞外 囊泡(EVS)或组织纳米转基因(TNT)为重编程提供了安全和微创的方法 体细胞和研究人员最近的工作表明，成体细胞成功地重新编程 成纤维细胞转化为内皮细胞的体内小鼠模型。我们建议使用这些创新的非病毒传递 将基因货物运送到体外和体内IVD的系统。第一个具体目标(R61阶段1目标1)重点 EV或TNT转导转录因子对人髓核病变细胞的影响 组织体外检测细胞外基质及分解代谢、炎症和疼痛标志物的变化。第二个目标(R61 阶段1目的2)研究EV或TNT转导转录因子在小鼠IVD穿刺术中的作用 评估椎间盘结构/功能、疼痛、认知和细胞毒性变化的DBP模型。这些研究 都是重要的和高度创新的，因为他们结合了一个独特的多学科医疗团队 以及兽医临床医生、脊柱生物学家、神经科学家、生物医学工程师和一名生物统计学家 询问如何使用这些新的概念和技术来治疗DBP。它的更广泛的影响 建议和过渡到R33部分涉及评估该策略在临床上的相关性 自发发生DBP的软骨营养不良犬，随后在DBP患者犬身上进行临床试验。

项目成果

Non-viral Gene Delivery Methods for Bone and Joints.

• DOI: 10.3389/fbioe.2020.598466
• 发表时间: 2020
• 期刊: Frontiers in bioengineering and biotechnology
• 影响因子: 5.7
• 作者: Gantenbein B;Tang S;Guerrero J;Higuita-Castro N;Salazar-Puerta AI;Croft AS;Gazdhar A;Purmessur D
• 通讯作者: Purmessur D

Non-viral reprogramming of human nucleus pulposus cells with FOXF1 via extracellular vesicle delivery: an in vitro and in vivo study.

• DOI: 10.22203/ecm.v041a07
• 发表时间: 2021-01-19
• 期刊: European cells & materials
• 影响因子: 3.1
• 作者: Tang S;Salazar-Puerta A;Richards J;Khan S;Hoyland JA;Gallego-Perez D;Walter B;Higuita-Castro N;Purmessur D
• 通讯作者: Purmessur D