Intranasal Delivery of Telomerase Reverse Transcriptase mRNA for Therapy ofTraumatic Brain Injury

鼻内递送端粒酶逆转录酶 mRNA 用于治疗创伤性脑损伤

基本信息

批准号：
10602034
负责人：
Biana Godin
金额：
$ 44.41万
依托单位：
METHODIST HOSPITAL RESEARCH INSTITUTE
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：
2022-09-26 至 2024-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10602034
关键词：
Address Affect Aging Animal Model Animals Apoptosis Area Attention Biodistribution Biological Availability Biomedical Engineering Blood flow Brain Brain Diseases Brain Injuries Brain region Bypass COVID-19 Catalytic Domain Cause of Death Cell Death Cell division Cells Central Nervous System Diseases Cessation of life Chromosome Structures Chromosomes Chronic Clinical Cognition DNA Damage DNA Repair DNA Sequence Data Degenerative Disorder Development Diagnostic Diffusion Drug Transport Effectiveness Enzymes Epigenetic Process Evaluation Event Functional disorder Head Health Care Costs Hour Immunologic Markers Impairment In Vitro Inflammation Injury Interruption Intranasal Administration Knowledge Label Length Link Longevity Longitudinal Studies Luciferases Measures Medical Messenger RNA Methodology Modeling Modification Morphology Motor Mus Nerve Degeneration Neurodegenerative Disorders Neurosciences Research Organ Outcome Oxidative Stress Patients Performance Persons Pharmaceutical Preparations Pharmacology Physiological Play Population Process Prognosis Property Proteins Psyche structure Quality of life RNA RNA vaccine RNA-Directed DNA Polymerase Regenerative capacity Reporter Reporter Genes Reporting Reproducibility Research Role Route System TBI treatment TERT gene Telomerase Telomere Shortening Testing Therapeutic Thinness Tissues Transfection Traumatic Brain Injury United States Untranslated RNA Vaccines Work base behavior test biomaterial compatibility brain cell brain tissue cell injury cerebrovascular cognitive ability cognitive disability cognitive function controlled cortical impact cyanine dye 5 design disability effective therapy experience gene therapy genetic information high reward high risk improved improved outcome in vivo in vivo Model innovation insight lipid nanoparticle mRNA Expression mRNA Transcript Degradation microfluidic technology mouse model multidisciplinary nanoparticle delivery nervous system disorder neuroinflammation neuron apoptosis neuronal survival novel novel therapeutics pandemic disease prevent programs recruit senescence synergism telomere tissue regeneration tissue repair tool uptake

项目摘要

ABSTRACT It is estimated that annually ~3 million Traumatic brain injury (TBI) cases occur in the U.S. Moderate to severe TBI can cause significant impairments in mental and motor functions or death. There are no effective therapies to improve cognitive abilities after moderate-severe TBI. Many novel pharmacological approaches for TBI therapy are not effective enough. Gene therapy can provide an unmet solution in protecting against several neurodegenerative disorders, including TBI. Recent advances in mRNA therapeutics/ vaccines have drawn significant attention due to their ability to tackle unmet clinical needs. For instance, the prompt development of COVID-19 mRNA vaccines aided in controlling the pandemics worldwide. Efficient mRNA therapies require a Lipid Nanoparticles (LNP) carrier to protect mRNA from degradation. Telomeres, repetitive non-coding DNA sequences, have a pivotal role in tissue repair and aging. Telomere shortening in the brain results from blood flow impairment and cell-death related inflammation and affects tissue regeneration ability. A catalytic subunit of telomerase, an enzyme responsible for maintaining telomere length (TL) during cell division, is telomerase reverse transcriptase (TERT). TL dysfunction has been implicated in neuroinflammatory and neurodegenerative processes and proposed as a marker for TBI outcomes. Additionally, TERT was shown to be important in neuronal survival and cognition, protecting from oxidative stress and blocking neuronal apoptosis. While TERT is a potential target in neurological disorders, no studies evaluating RNA therapy to address TL in TBI were reported yet. Here we propose a transformational therapeutic approach for TBI therapy which includes intranasal (IN) TERT mRNA- LNP delivery to the brain. LNP protect mRNA en route to the target TBI tissue. The immediate focus of our work is an impairment in the brain's normal function caused by an impact to the head. Our data show that IN delivery of LNP bypasses BBB, enhancing drug transport to the brain. Our teams have demonstrated that TERT mRNA can enhance TL in vitro and in vivo improving prognosis in other degenerative conditions. We have also shown that there is a shortening of telomeres and reduction in TERT levels in our TBI mouse model. Our approach may radically change therapy for TBI, as well as other brain disorders. In this exploratory project, our central hypothesis is that IN administration of TERT mRNA will enable temporary expression of TERT in the affected brain tissue restoring cognitive functions after TBI. Our Specific aims are: (1) Design, characterization, and biocompatibility of mRNA-LNP in vitro: four TERT-mRNA-LNP systems will be designed, characterized, and assessed in vitro with various brain cells; (2) Evaluate the biodistribution and therapeutic efficiency of IN administration of TERT mRNA-LNPs in a mouse model of TBI (Controlled cortical impact, CCI). Biodistribution of reporter mRNA and fluorescently labeled LNP in the brains of mice with TBI will be performed. Efficiency will be tested based on TERT levels, TL, behavioral tests, and immunological markers. This exploratory project can be the first step in the development of a much-needed therapy for TBI.

抽象的据估计，美国每年发生约 300 万例创伤性脑损伤 (TBI) 病例中度至重度 TBI 可导致精神和运动功能严重受损或死亡。没有有效的改善中重度 TBI 后认知能力的疗法。许多新颖的药理学方法 TBI 治疗不够有效。基因疗法可以提供一个未满足的解决方案来预防多种疾病神经退行性疾病，包括 TBI。 mRNA 疗法/疫苗的最新进展吸引了由于其解决未满足的临床需求的能力而受到极大关注。例如，迅速发展 COVID-19 mRNA 疫苗有助于控制全球流行病。有效的 mRNA 疗法需要脂质纳米颗粒 (LNP) 载体可保护 mRNA 免遭降解。端粒，重复的非编码DNA 序列，在组织修复和衰老中具有关键作用。血液导致大脑端粒缩短血流障碍和细胞死亡相关的炎症并影响组织再生能力。催化亚基端粒酶，一种在细胞分裂过程中负责维持端粒长度（TL）的酶，是端粒酶逆转录酶（TERT）。 TL 功能障碍与神经炎症和神经退行性病变有关过程并建议作为 TBI 结果的标记。此外，TERT 被证明在以下方面也很重要：神经元存活和认知，防止氧化应激并阻止神经元凋亡。而叔是神经系统疾病的潜在靶点，目前还没有评估 RNA 疗法解决 TBI 中 TL 的研究尚未报道。在这里，我们提出了一种 TBI 治疗的变革性治疗方法，其中包括鼻内治疗 (IN) TERT mRNA-LNP 递送至大脑。 LNP 可保护 mRNA 到达目标 TBI 组织。即时的我们工作的重点是头部撞击造成的大脑正常功能受损。我们的数据表明 LNP 的 IN 递送绕过 BBB，增强药物向大脑的转运。我们的团队有证明 TERT mRNA 可以在体外和体内增强 TL，改善其他退行性疾病的预后状况。我们还表明，我们的 TBI 中存在端粒缩短和 TERT 水平降低的情况鼠标模型。我们的方法可能会从根本上改变 TBI 以及其他脑部疾病的治疗。在这个探索性项目，我们的中心假设是 TERT mRNA 的 IN 管理将能够暂时 TERT 在受影响的脑组织中的表达可恢复 TBI 后的认知功能。我们的具体目标是： (1) mRNA-LNP 体外设计、表征和生物相容性：将开发四个 TERT-mRNA-LNP 系统用各种脑细胞进行体外设计、表征和评估； (2) 评估生物分布和在 TBI 小鼠模型中 IN 施用 TERT mRNA-LNP 的治疗效果（受控皮质影响，CCI）。报告基因 mRNA 和荧光标记 LNP 在小鼠大脑中的生物分布 TBI 将被执行。将根据 TERT 水平、TL、行为测试和免疫学标记来测试效率。这个探索性项目可能是开发急需的 TBI 疗法的第一步。