Novel Gene Therapy for Critical Limb Ischemia and Limb Salvage in Vascular Occlusive Disease

针对血管闭塞性疾病严重肢体缺血和保肢的新型基因疗法

• 批准号: 10447520
• 负责人: ZHAO-JUN LIU
• 金额: $ 53.33万
• 依托单位: UNIVERSITY OF MIAMI SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-05-24 至 2023-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10447520
• 关键词: Amputation; Angiogenic Factor; Angioplasty; Animal Experimentation; Animals; Arteries; Atherectomy; Atherosclerosis; Biodistribution; Biological; Biological Assay; Blood; Blood Tests; Blood Vessels; Blood capillaries; Blood flow; Bypass; Caring; Cell Adhesion Molecules; Cell Communication; Cell physiology; Cells; Cellular Morphology; Clinical; Clinical Trials; Data; Development; Disease; Docking; Dose; E-Selectin; Endothelial Cells; Face; Fibrin fragment D; Fibroblast Growth Factor; Foundations; Funding; Future; Gangrene; Gene Delivery; Generations; Goals; Harvest; Health Care Costs; Homing; Human; In Vitro; Inflammation; Intervention; Ischemia; Isolated limb perfusion; Ligands; Limb Salvage; Limb structure; Long-Term Effects; Mechanics; Medical; Metabolic; Modeling; Mus; Muscle Cells; Muscle Fibers; National Heart, Lung, and Blood Institute; Natural regeneration; Organ; Pain; Pain in lower limb; Paracrine Communication; Pathologic; Patients; Peripheral arterial disease; Phase; Physical Function; Pilot Projects; Plasmids; Process; Protocols documentation; Recovery; Renal function; Research; Rest; Running; Safety; Salvage Therapy; Serum; Site; Skeletal Muscle; Stains; Stents; Structure; Testing; Therapeutic; Therapeutic Effect; Thrombosis; Tissues; Toxic effect; Transgenes; Tumor-infiltrating immune cells; Vascular Endothelial Growth Factors; Viral; Walking; Work; adenoviral-mediated; angiogenesis; artery occlusion; base; clinically relevant; critical limb Ischemia; delivery vehicle; efficacy validation; endothelial stem cell; experience; gene delivery system; gene therapy; healing; high risk; human study; improved; in vivo; innovation; intercellular communication; limb amputation; limb ischemia; limb loss; liver function; mortality risk; mouse model; neovascularization; non-healing wounds; novel; novel therapeutics; preclinical study; prevent; progenitor; recruit; regenerative; regenerative cell; repaired; scale up; skin ulcer; stability testing; stem; stem cells; success; systemic toxicity; therapeutic angiogenesis; tissue regeneration; tissue repair; translational research program; treadmill; treatment optimization; vector

Project Summary/Abstract: Critical limb ischemia (CLI) is the most advanced stage of peripheral arterial disease (PAD). It manifests with leg pain, decreased walking distance, non-healing wounds and gangrene. About 1/3 of CLI patients don't respond to standard therapy and ultimately undergo major limb amputation. This critical unmet need demands a solution. CLI causes >105 major limb amputations in the USA per year. Our overarching goal is to develop a novel limb-salvage therapy. Gene therapy is a promising option to induce therapeutic angiogenesis, increasing blood flow in ischemic limbs for repair. This approach has raised a great deal of hope for `no-option' CLI patients. However, previous gene therapy approaches using naked plasmids encoding angiogenic factors, such as VEGF and FGF, failed to show substantial clinical benefits. Moreover, early adenovirus-mediated gene delivery systems raised concerns over safety. Hence, more efforts for the identification of alternative effective transgenes and safe/efficient delivery vectors are crucial. Success of gene therapy for CLI requires activation and participation of endogenous healing mechanisms including recruitment/homing of stem/progenitor cells to ischemic tissue. Enhanced recruitment/homing depends upon a `hospitable' tissue microenvironment that is receptive of endogenous regenerative cells. Our previous research discovered that in ischemia, homing for these critical repair cells relies on specific cell-cell interactions via a panel of adhesion molecules, including E- selectin/ligand pairs. The levels of vascular E-selectin in the ischemic limb tissues is down-regulated. We hypothesize that E-selectin is pivotal in the recruitment of stem/progenitor cells necessary for neovascularization in ischemic tissues. E-selectin can serve as a docking site for endogenous stem/progenitor cells to anchor. The resulting downstream cell-cell signaling cascades can increase angiogenesis and tissue repair/regeneration. We have recently demonstrated feasibility, safety and superior efficacy of E-selectin/AAV- based gene therapy for improving limb perfusion, increasing neovascularization, decreasing ischemia, reducing gangrene, minimizing tissue damage, augmenting regeneration of the ischemic skeletal muscle, and extending running capacity, in murine models. In the clinical setting, we believe that such biologic effects can increase functional walking capacity and prevent limb loss. We thus propose to induce a supportive tissue microenvironment by priming endothelial cells in capillaries and other ischemic limb-tissue cells with the adhesion molecule, E-selectin, using a safe and efficient AAV2 vector. We will utilize a clinically relevant mouse model of CLI – the footpad gangrene, to test this novel gene therapy. The objectives are to validate and optimize the translational protocol for a future large-animal pre-clinical study and conduct the requisite IND-enabling GLP/GMP animal research for a subsequent pilot human study. The development of this novel gene therapy may ultimately eliminate the need for most amputations by accelerating therapeutic angiogenesis and tissue regeneration, and promises to revolutionize the treatment for PAD/CLI patients.

项目摘要/摘要： 严重肢体缺血(CLI)是外周动脉疾病(PAD)的最晚期。它表现为 腿部疼痛、步行距离缩短、无法愈合的伤口和坏疽。大约三分之一的CLI患者没有 对标准治疗有反应，并最终接受主要肢体截肢。这一关键的未得到满足的需求 一个解决方案。CLI每年在美国造成105例大腿截肢。我们的首要目标是开发一种 新的保肢疗法。基因治疗是诱导治疗性血管生成的一个有前途的选择，增加了 供修复的缺血肢体的血液流动。这种方法为“没有选择”的CLI患者带来了极大的希望。 然而，以前的基因治疗方法使用编码血管生成因子的裸质粒，例如 血管内皮细胞生长因子和成纤维细胞生长因子未能显示出实质性的临床疗效。此外，早期的腺病毒介导的基因传递 这些系统引发了人们对安全的担忧。因此，更多的努力寻找有效的替代方案 转基因和安全/高效的递送载体至关重要。CLI基因治疗的成功需要激活 以及内源性修复机制的参与，包括干细胞/祖细胞的招募/归巢 缺血组织。增强招募/归巢取决于组织微环境的好客， 内源性再生细胞的接受性。我们之前的研究发现，在缺血状态下，归巢 这些关键的修复细胞依赖于通过一组黏附分子进行的特定的细胞间相互作用，包括E- 选择素/配体对。缺血肢体组织中血管E-选择素水平下调。我们 假设E-选择素在干细胞/祖细胞的募集中起关键作用 缺血组织中的新生血管。E-选择素可作为内源性干/祖细胞的对接部位 要锚定的细胞。由此产生的下游细胞-细胞信号级联可以增加血管生成和组织 修复/再生。我们最近已经证明了E-选择素/AAV的可行性、安全性和优越的疗效。 基于基因治疗的改善肢体血流，增加新生血管，减少缺血，减少 坏疽，最大限度地减少组织损伤，增强缺血骨骼肌的再生，并延伸 跑步能力，在小鼠模型中。在临床环境中，我们相信这样的生物效应可以增加。 功能行走能力，防止肢体丧失。因此，我们建议诱导一种支持组织 微环境通过用血管内皮细胞和其他缺血肢体组织细胞 黏附分子，E-选择素，使用安全有效的AAV2载体。我们将利用一种临床相关的小鼠 CLI模型-足部坏疽，以测试这一新的基因疗法。目标是验证和优化 未来大型动物临床前研究的翻译方案，并进行必要的IND使能 GLP/GMP动物研究，用于随后的先导性人体研究。这种新型基因疗法的研究进展 可能最终通过加速治疗性血管生成和组织来消除大多数截肢的需要 再生，并承诺将彻底改变PAD/CLI患者的治疗方法。