Ultrasound-guided DNA delivery for regenerative medicine

用于再生医学的超声引导 DNA 递送

基本信息

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
项目类别：
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

项目摘要

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种体内超声介导的核酸靶向再生系统作为骨骼组织再生方面的专家，我们打算提供骨科损伤的概念证明。首先，骨折不愈合和肌腱/韧带撕裂模型尚未满足临床需求。骨科医学并需要开发新的治疗方法来治疗不愈合骨折。韧带撕裂包括自体移植和同种异体移植，这些都涉及严重的并发症和长期的治疗。骨形态发生蛋白（BMP）基因传递，通过使用来完成愈合和丧失活动能力的时期。病毒已被证明可以在啮齿动物和大型动物中诱导骨折不愈合。载体是最有效的基因传递工具，但它们也带来了潜在的致瘤风险和非病毒载体被认为对人类使用更安全，但对人类使用效率较低。基因表达的另一种物理方法称为声孔法。由于当今超声在临床中的广泛使用，它对临床应用很有吸引力。采用局部注射微泡以及随后的超声成像和治疗来实现高最近我们能够证明基于超声的 BMP-6 基因。在小型猪模型中，分娩实现了完整的节段性骨缺损修复（科学转化医学， 2017）此外，还使用了类似的方法将质粒 DNA 输送到为小型猪膝关节的韧带重建目前存在三个主要障碍。声孔在骨科手术中的应用： 1. 节段性骨折部位具有独特的金属- 骨-软组织界面，造成超声波反射强烈；2.超声波难以瞄准；脉冲进入狭窄的骨隧道以实现肌腱/韧带移植骨整合，以及 3. 空化的控制活动也是创新和临床进步的障碍和机会。依靠费拉拉集团在超声控制药物输送领域的广泛专业知识来克服克服上述障碍，推动超声介导的骨骼再生工程走向临床。我们提出两个目标：目标 1：针对临界尺寸骨折设计并测试图像引导转染。目标：1A) 开发传感器和软件以在 3D 体积内扫描 US 波束，同时进行测绘和控制空化活动目标：1B) 在猪临界尺寸骨折模型中测试骨折愈合的功效。目标 2：设计并测试针对韧带/肌腱骨隧道定制的图像引导转染。目标：2A) 开发传感器和软件，在测绘和控制的同时在骨隧道内引导超声束 2B) 在猪韧带重建模型中测试增强修复的功效。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。