Ultrasound image-guided treatment of ischemia-reperfusion injury using argon microbubbles

超声图像引导氩气微泡治疗缺血再灌注损伤

• 批准号: 10303690
• 负责人: Daeyeon Lee
• 金额: $ 8.13万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-01 至 2023-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10303690
• 关键词: 1-Phosphatidylinositol 3-Kinase; 3-Dimensional; Acoustics; Acute; Animal Model; Animals; Applications Grants; Argon; Biological Assay; Biological Markers; Blood Circulation; CASP3 gene; Cardiac; Caspase; Cell Culture Techniques; Cell Line; Cell Survival; Cell membrane; Cells; Centrifugation; Clinic; Clinical; Collaborations; Colloids; Cytoprotective Agent; Development; Encapsulated; Engineering; Ensure; Environment; Exhibits; Exposure to; Formulation; Future; Gases; Glucose; Glutamate Receptor; Goals; Health; Heart Arrest; Heart Injuries; Hypoxia; In Vitro; Incentives; Inhalation; Injury; Kidney; Length; Lipids; Liposomes; Literature; Measures; Mediating; Methods; Microbubbles; Microfluidics; Modeling; Myocardial Infarction; Neuronal Injury; Neurons; Nitric Oxide; Noble Gases; Oxidative Stress; Oxygen; Pathway interactions; Pediatric Hospitals; Perfusion; Philadelphia; Phospholipids; Physiological; Preclinical Testing; Preparation; Production; Property; Proto-Oncogene Proteins c-akt; Protocols documentation; Reperfusion Injury; Reperfusion Therapy; Reporting; Research; Rest; Signal Pathway; Signal Transduction; Site; Solubility; Sonication; Stroke; System; Testing; Therapeutic; Therapeutic Effect; Time; Tissues; Translating; Translations; Traumatic Brain Injury; Ultrasonography; Up-Regulation; Validation; Work; Xenon; aqueous; base; clinical application; deprivation; design; disability; early phase clinical trial; efficacy evaluation; efficacy testing; experience; hypoxic ischemic injury; image guided; image guided therapy; improved; in vivo; in vivo imaging; interfacial; ischemic injury; mouse model; non-invasive imaging; prevent; scale up; side effect; simulation; theranostics; treatment effect; validation studies

The key health significance of this proposal involves the ultrasound-mediated, image-guided, localized treatment of ischemia reperfusion injury (IRI) in neuronal and cardiac models using echogenic argon microbubbles (ArMBs). There exist no clinically approved methods for treating damaged tissue after experiencing hypoxic ischemic and reperfusion injuries such as stroke or cardiac arrest. Noble gases like argon (Ar) and xenon (Xe) are highly promising cytoprotective agents that have been shown to successfully treat acute IRI in vitro and in animal models. Whereas Xe has been researched in greater detail including in early clinical trials, it can be prohibitively expensive and difficult to obtain. Ar is a hundred times cheaper and widely available, while exhibiting excellent organoprotective efficiency. Furthermore, the mechanism of Xe action depends on its interaction with glutamate receptors on cell membranes, whereas Ar is reported to work by stimulating various endogenous cellular protecting signaling pathways, making it a more versatile antiapoptotic agent. Current Ar therapy is long and systemic, via inhalation, making it non-specific to the injury site, likely diminishing therapeutic effect. As a solution, we propose the development of MBs (MBs) for localized delivery of the therapeutic gas. MBs are inherently echogenic due to their non-linear oscillations induced by clinical ultrasound. Therapeutic gases such as Ar, however, are difficult to stabilize inside bubbles due to the former's high aqueous solubility. The team has recently succeeded in small-scale production of stable, echogenic, noble gas MBs through optimization of the MB shell composition, leading to a productive, ongoing collaboration with clinicians at the Children's Hospital of Philadelphia (CHOP). The proposed research will be conducted through the implementation of three specific aims. (1) 1-10 µm ArMBs will be formulated at a high yield of >1010 MBs per mL. Ultrasound signal of optimized ArMBs will be investigated in flow phantoms and in a mouse model by measuring the magnitude, perfusion, and persistence of contrast. (2) The therapeutic effect of ultrasound mediated Ar release from bubbles in treating IRI will be estimated in in vitro cell culture-based simulations of neuronal and cardiac injuries induced by oxygen glucose deprivation. The validity of ArMBs will be proved by enhanced cell viability, decrease in caspase activation, and upregulation in the phosphatidylinositol 3 kinase (PI3K-AKT) pathway. (3) Further incentive to use ArMBs will be recognized by comparing their IRI treatment results to that of bulk Ar exposure to cells and exposure to XeMBs. ArMB activity even with the deactivation of glutamate receptors will be shown to cement the feasibility of ArMBs for a variety of cytoprotective treatments. The PI team will leverage their expertise in colloidal design, cellular dynamics, and ultrasound imaging to precisely engineer the ArMB shell and to rigorously establish the validity of this new, inexpensive agent in vitro for the team's long-term goal of testing ArMBs for non-invasive, image guided treatment of IRI in large animal models and translating them to clinical settings.

这项建议的关键健康意义涉及超声介导的、图像引导的局部治疗 利用回声微泡对神经元和心脏缺血再灌注损伤(IRI)的影响 (ARMBs)。目前尚无临床认可的治疗缺氧后受损组织的方法。 缺血和再灌注损伤，如中风或心脏骤停。惰性气体，如Ar(Ar)和Xe(Xe) 是非常有希望的细胞保护剂，已被证明在体外和在 动物模型。尽管已经对Xe进行了更详细的研究，包括在早期临床试验中，但它可以 昂贵得令人望而却步，而且很难获得。AR的价格要便宜一百倍，而且可以广泛获得，同时展示了 极佳的有机保护效率。此外，Xe的作用机制依赖于它与 细胞膜上的谷氨酸受体，而AR被报道通过刺激各种内源性 保护细胞信号通路，使其成为一种更通用的抗凋亡剂。目前的Ar疗法是长期的 全身，通过吸入，使其对损伤部位没有特异性，可能会降低治疗效果。作为一名 为了解决这一问题，我们建议开发用于局部输送治疗性气体的MBS(MBS)。MBS是 由于临床超声引起的非线性振荡，导致固有的回声。治疗气体，如 然而，由于Ar在水中的高溶解度，Ar很难稳定在气泡中。这支队伍有 最近通过优化工艺，成功地小规模生产了稳定的、回声的、惰性气体MBS MB壳成分，导致与儿童医院的临床医生进行富有成效的持续合作 费城(CHOP)。拟议的研究将通过实施三个具体的 目标。(1)1-10微米ARMBs将以每毫升1010 MBS的高产量配制。优化后的超声信号 将在流动模体和小鼠模型中通过测量大小、灌注量和 对比度的持续性。(2)超声介导Ar气泡释放治疗IRI的疗效观察 将在体外细胞培养基础上模拟氧引起的神经元和心脏损伤进行评估 葡萄糖缺乏。ARMBs的有效性将通过提高细胞存活率、降低caspase来证明。 磷脂酰肌醇3激酶(PI3K-AKT)的激活和上调。(3)进一步鼓励 使用ARMBs将通过将其IRI治疗结果与大量暴露于细胞和 暴露在XeMBs中。ArMB活性即使在谷氨酸受体失活的情况下也会显示为粘结剂 ARMBs用于多种细胞保护治疗的可行性。PI团队将利用他们的专业知识在 胶体设计、细胞动力学和超声成像，以精确设计ArMB壳并严格地 确定这种新的、廉价的体外试剂的有效性，以实现该团队测试ARMBs的长期目标 在大型动物模型中对IRI进行非侵入性、图像引导的治疗，并将其转化为临床环境。