Ultrasound-guided DNA delivery for regenerative medicine

用于再生医学的超声引导 DNA 递送

• 批准号: 10113360
• 负责人: DAN GAZIT
• 金额: $ 69.09万
• 依托单位: CEDARS-SINAI MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-05-01 至 2022-06-16
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10113360
• 关键词: 3-Dimensional; Allografting; Animal Model; Animals; Auditory; Autologous Transplantation; BMP6 gene; Bone Injury; Bone Morphogenetic Proteins; Bone Regeneration; Brain; Cell Nucleus; Clinic; Clinical; Complex; Computer software; Custom; DNA; DNA delivery; Defect; Development; Diagnostic Imaging; Drug Delivery Systems; Electroporation; Engineering; Family suidae; Fracture; Fracture Healing; Frequencies; Gene Delivery; Gene Expression; Genes; Goals; Heart; Human; Injections; Injury; Knee joint; Ligaments; Lithotripsy; Location; Lung; Mediating; Medicine; Mesenchymal Stem Cells; Metals; Methods; Microbubbles; Miniature Swine; Modeling; Monitor; Non-Viral Vector; Nucleic Acids; Ocular orbit; Orthopedic Surgery; Orthopedics; Osseointegration; Osteogenesis; Patients; Physiologic pulse; Plasmids; Quality of life; Reaction; Regenerative Medicine; Regenerative engineering; Reporter Genes; Reporting; Research; Risk; Rodent; Rotator Cuff; Site; Skeleton; Source; Structure; System; Technology; Tendon structure; Testing; Transducers; Transfection; Translational Research; Traumatic injury; Ultrasonic Therapy; Ultrasonics; Ultrasonography; Viral Vector; Virus; anterior cruciate ligament reconstruction; anterior cruciate ligament rupture; base; body system; bone; bone loss; cancer therapy; clinical application; clinically relevant; efficacy testing; healing; image guided; imaging system; immunogenic; in vivo; innovation; long bone; novel; novel therapeutics; nucleic acid delivery; overexpression; plasmid DNA; porcine model; pre-clinical; prevent; reconstruction; repaired; side effect; signal processing; skeletal; skeletal tissue; soft tissue; sonoporation; stem cells; tissue regeneration; tool; translational medicine; tumor ablation; tumorigenic

ABSTRACT We propose to develop a system of in vivo ultrasound-mediated nucleic acid targeting for regenerative medicine. As experts in skeletal tissue regeneration, we intend to provide proof of concept in orthopedic injury models, first. Nonunion bone fractures and tendon/ligament tears are unmet clinical needs in the field of orthopedic medicine and require the development of novel therapies. Current treatments to nonunion fractures and ligament tears include autografts and allografts, which all involve serious complications and prolonged periods of healing and loss of mobility. Bone Morphogenetic Protein (BMP) gene delivery, accomplished using viruses, has been shown to induce healing of nonunion fractures in rodents and large animals. While viral vectors are the most efficient gene delivery tools, they also introduce potential risks of tumorigenic and immunogenic reactions. Nonviral vectors are considered safer for human use, albeit much less efficient for gene expression. An alternative physical method of gene transfection termed sonoporation is especially attractive for clinical applications due to the widespread use of ultrasound in the clinic today. Here, we will employ local injection of microbubbles and subsequent ultrasound imaging and therapy to achieve high efficiency local transfection. Recently we were able to demonstrate that ultrasound-based BMP-6 gene delivery yielded complete segmental bone defect repair in a mini-pig model (Science Translational Medicine, 2017). Furthermore, a similar approach was used to deliver plasmid DNA to bone tunnels created for the ligament reconstruction in a minipigs knee joints. However, currently three main hurdles prevent widespread implementation of sonoporation for orthopaedic surgery: 1. The segmental fracture site poses a unique metal- bone-soft tissue interface, which causes strong ultrasound reflections; 2. It is difficult to target the ultrasonic pulse into narrow bone tunnels for tendon/ligament graft osseointegration, and 3. The control of cavitation activity is also a hurdle and an opportunity for innovation and clinical advancements. Here we propose to rely on the extensive expertise of the Ferrara group in the field of ultrasound–controlled drug delivery to overcome the abovementioned hurdles and promote ultrasound mediated skeletal regenerative engineering to the clinic. We propose two aims: Aim 1: Engineer and test image-guided transfection for critical-size bone fractures. Goal: 1A) Develop transducer and software to sweep the US beam within a 3D volume while mapping and controlling cavitation activity. Goal: 1B) Test the efficacy of fracture healing in a pig critical-size fracture model. Aim 2: Engineer and test image-guided transfection customized for ligament/tendon bone tunnels. Goal: 2A) Develop transducer and software to direct the US beam within a bone tunnel while mapping and controlling cavitation activity. 2B) Test the efficacy of enhanced repair in a pig ligament reconstruction model. If successful, this project could be highly beneficial for numerous clinical applications beyond orthopedic targets, including cancer therapy.

抽象的 我们建议开发一个体内超声介导的核酸靶向重生系统 药品。作为骨骼组织再生的专家，我们打算提供骨科损伤的概念证明 模型，首先。骨不骨骨折和肌腱/韧带撕裂是未满足的临床需求 骨科医学需要开发新疗法。当前对骨不骨骨折的治疗 韧带眼泪包括自体移植和同种异体移植物，所有这些都涉及严重的并发症并延长 康复期和流动性丧失。骨形态发生蛋白（BMP）基因递送，使用 病毒已被证明可诱导啮齿动物和大动物中骨不连断裂的愈合。虽然病毒 向量是最有效的基因输送工具，它们还引入了肿瘤症和 免疫原性反应。非病毒载体被认为对人类的使用更安全，尽管效率较低 基因表达。尤其是称为somotoration的基因转染的另一种物理方法是 由于当今诊所的超声使用宽度，对临床应用有吸引力。在这里，我们会的 员工局部注入微泡以及随后的超声成像和治疗，以达到高 效率本地翻译。最近，我们能够证明基于超声的BMP-6基因 输送在迷你铅模型中产生了完全节骨缺损修复（科学翻译医学， 2017）。此外，使用类似的方法将质粒DNA输送到为骨隧道提供的 韧带重建在膝关节关节中。但是，目前的三个主要障碍阻止了宽度 实施骨科手术的超声层：1。节段骨折部位构成独特的金属 - 骨软组织界面，会引起强大的超声反射； 2。很难针对超声波 将肌腱/韧带移植骨整合的狭窄骨隧道脉冲，3。 活动也是创新和临床进步的障碍和机会。我们建议在这里依靠 在超声控制的药物输送领域，费拉拉集团的广泛专业知识以克服 上述障碍并促进了诊所的超声介导的骨骼再生工程。 我们提出了两个目的：目标1：工程师和测试图像引导的翻译，用于临界大小的骨折。 目标：1A）开发传感器和软件，以在映射和映射时将美国光束扫荡在3D卷中 控制空化活动。目标：1B）测试猪临界大小骨折模型中断裂愈合的效率。 AIM 2：针对韧带/肌腱骨隧道定制的工程师和测试图像引导的翻译。目标：2a） 开发换能器和软件，以在绘制和控制的同时将美国光束引导到骨隧道内 空化活动。 2b）测试猪韧带重建模型中增强修复的效率。如果 成功，该项目可能对骨科目标以外的众多临床应用非常有益， 包括癌症治疗。