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.

摘要 我们建议开发一种体内超声介导的核酸靶向系统， 药作为骨骼组织再生领域的专家，我们打算为骨科损伤提供概念验证 模特们先来骨不连骨折和肌腱/韧带撕裂是外科领域中未满足的临床需求。 整形外科医学和需要开发新的疗法。骨折不愈合的治疗现状 和韧带撕裂包括自体移植和同种异体移植，这都涉及严重的并发症和长期 恢复期和丧失活动能力期。骨形态发生蛋白（BMP）基因递送，使用 病毒，已被证明可以诱导啮齿动物和大型动物的骨折不愈合。而病毒 载体是最有效的基因递送工具，它们也引入了潜在的致瘤风险， 免疫原性反应非病毒载体被认为对人类使用更安全，尽管对人类使用的效率要低得多。 基因表达。另一种称为声致穿孔的基因转染物理方法， 由于当今超声在临床中的广泛使用，因此对于临床应用是有吸引力的。在这里，我们将 采用局部注射微泡和随后的超声成像和治疗， 高效局部转染。最近，我们能够证明基于超声的BMP-6基因 递送在小型猪模型中产生了完全的节段性骨缺损修复（ScienceTranslationalMedicine， 2017年）。此外，使用类似的方法将质粒DNA递送到为骨形成的骨隧道中。 小型猪膝关节韧带重建术。然而，目前有三个主要障碍阻止了广泛的 声致孔在骨科手术中应用：1.节段性骨折的部位有独特的金属- 骨-软组织界面，其引起强超声反射; 2.很难瞄准超声波 脉冲进入狭窄的骨隧道，用于肌腱/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。在这里，我们建议依靠 凭借费拉拉集团在超声控制药物输送领域的广泛专业知识， 并将超声介导的骨再生工程推向临床。 我们提出了两个目标：目标1：工程师和测试图像引导转染临界尺寸的骨折。 目标：1A）开发探头和软件，以便在标测时在3D体积内扫描US射束， 控制空化活动。目的：1B）在猪临界尺寸骨折模型中测试骨折愈合的功效。 目标2：设计和测试为韧带/肌腱骨隧道定制的图像引导转染。目标：2A） 开发探头和软件，在标测和控制的同时，引导骨隧道内的超声束 空化活动2B）在猪韧带重建模型中测试增强修复的功效。如果 如果成功，该项目可能对骨科目标以外的许多临床应用非常有益， 包括癌症治疗。