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 慢性下腰痛为社会带来了重要的社会经济伯恩，是导致该社会的主要贡献者 增长阿片类药物危机。去年，NIH“启动了治愈（有助于长期结束成瘾）倡议，这是 积极的，跨机构的努力，以加快科学解决方案来阻止国家阿片类药物公共卫生危机。” 这种巨大的伯宁主要是因为研究未能针对相关的潜在机制 产生疼痛。椎间盘（IVD）变性与病理生理密切相关 下背痛并识别出非添加性的微创腰痛治疗方法（DBP）是 研究优先。病理IVD变化包括细胞外基质（ECM）分解，注射和 异常的神经/血管内膜已显示与疼痛显着相关。因此 DBP的最佳治疗将既靶向结构恢复并减轻疼痛的症状。但最新 涉及使用干细胞或基因治疗的策略面临许多挑战，包括 干细胞无法适应HARMH IVD微环境，使用病毒载体和不必要的DNA 宿主基因组中的删除。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患者的潜在疾病的技术。有急需 针对解决这些局限性的生物非添加策略。我们的目标是使用新颖的蜂窝 重编程技术以将天然解散的IVD细胞的先天细胞表型改变为健康 人体体外细胞培养和体内的细胞外产生和抗代谢/炎症表型 DBP的模型。非病毒输送，用于将遗传货物运输到细胞中，例如工程细胞外 囊泡（EV）或组织纳米转染（TNT）提供了安全且微创的方法用于重编程 调查人员的躯体细胞和最近的工作表明，成人的成功重编程 成纤维细胞进入体内小鼠模型中的内皮细胞。我们建议使用这些创新的非病毒交付 在体外和体内向IVD运送遗传货物的系统。第一个特定目的（R61阶段1 AIM 1）聚焦 关于转录因子EV或TNT递送对解剖的人核细胞和 在体外检查ECM和分解代谢，炎症和疼痛标记的变化。第二个目标（R61） 第1阶段目标2）研究小鼠IVD穿刺中转录因子的EV或TNT递送的影响 DBP评估模型的椎间盘结构/功能，疼痛，认知和细胞毒性的变化。这些研究 既重要又高度创新，因为它们结合了一个独特的医学多学科团队 以及兽医临床医生，脊柱生物学家，神经科学家，生物医学工程师和生物统计学家 询问这些新颖概念和技术来治疗DBP的使用。更广泛的影响 提案和向R33部分的过渡涉及评估该策略的临床相关策略 在患有DBP的患者狗中，发育于发育DBP的软骨型狗，然后进行临床试验。

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