Developing a Temporally-Regulated Gene Therapy for Therapeutic Angiogenesis

开发用于治疗性血管生成的时间调控基因疗法

• 批准号: 10535141
• 负责人: Mai Ngo
• 金额: $ 6.72万
• 依托单位: BOSTON UNIVERSITY (CHARLES RIVER CAMPUS)
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-30 至 2024-09-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10535141
• 关键词: Affect; Alternative Therapies; Angiogenic Factor; Angioplasty; Arteries; Behavior; Biocompatible Materials; Biological; Blood Vessels; Blood flow; Boston; Bypass; Cardiovascular Diseases; Cause of Death; Cell Therapy; Cells; Cessation of life; Chronic; Clinic; Clinical Trials; Complex; Development; Dose; Endothelial Cells; Engineering; Event; Gene Delivery; Gene Expression; Gene Expression Regulation; Generations; Genes; Genetic; Grant; Growth; Growth Factor; Growth Factor Gene; Half-Life; Health; Hindlimb; Hypoxia; Hypoxia Inducible Factor; Immunotherapy; Impairment; In Vitro; Ischemia; Maturation-Promoting Factor; Measures; Mentors; Modeling; Molecular; Operative Surgical Procedures; Oxygen; Patients; Perfusion; Persons; Play; Process; Proliferating; Quality of life; Recombinant Proteins; Recovery; Regenerative Medicine; Response Elements; Role; Signal Transduction; Site; Stroke; Switch Genes; Testing; Tissue Engineering; Tissues; Training; Transcriptional Regulation; Translating; Tube; Universities; Vascular blood supply; Vascularization; Writing; angiogenesis; career; cellular engineering; collaborative environment; comorbidity; cost; design; disability; experience; gene therapy; genetic element; genetically modified cells; improved; in vitro Model; in vivo; in vivo Model; insight; interest; ischemic cardiomyopathy; next generation; novel; paracrine; patient population; recruit; sensor; stem cells; success; symposium; synthetic biology; therapeutic angiogenesis; tool

Project Summary/Abstract Cardiovascular diseases affect millions of patients worldwide and account for nearly a third of deaths globally. Ischemia, or a reduced blood supply, occurs in many cardiovascular diseases and is a pressing health challenge. While current treatments primarily focus on re-vascularization of existing blood vessels, a significant sub- population of patients are unable to tolerate the associated surgical procedures due to existing comorbidities. Thus, there is great interest in developing strategies for therapeutic angiogenesis, which seeks to stimulate new vascularization at the ischemic site. While many gene and cell therapies for therapeutic angiogenesis have been tested in clinical trials, a clear benefit for patients remains to be seen. To date, most gene therapies deliver one or two genes to the ischemic site, while cell therapies deliver progenitor or stem cells to produce paracrine factors and self-organize into vasculature. A central limitation of these therapies is the inability to control the temporal presentation of the expressed genes or secreted factors. Angiogenesis is a complex and temporally regulated process, in which angiogenic factors first initiate the formation of a primitive vascular network before maturation factors promote mural cell recruit and vessel stabilization. While studies with growth factors suggest that sequential delivery of angiogenic and maturation factors is beneficial for establishing functional vasculature, how the timing of the angiogenic-to-maturation transition impacts the functionality of the established vasculature is unknown. How tissues naturally sense the correct timing for the angiogenic-to- maturation transition is also unclear, but incorporating a sensor to regulate the expression of angiogenic and maturation genes would be beneficial for creating a gene therapy with controlled dosing and minimal off-target effects. In this proposal, synthetic biology tools will be combined with engineered models of vascularization and an in vivo model of hindlimb ischemia to evaluate how the timing of angiogenic and maturation gene expression impacts functional vascular network formation and recovery from ischemia. In Aim 1, a two-channel genetic switch will be used to establish the relationship between the timing of the angiogenic-to-maturation transition and vascular network functionality. In Aim 2, hypoxia response elements will be used to generate a hypoxia-regulated genetic switch to control the induction of angiogenic and maturation genes. The genetic switch will be evaluated for its ability to rescue perfusion in an in vivo hindlimb ischemia model. The associated training plan will prepare the fellow for an academic career by enabling the fellow to obtain new skillsets in synthetic biology and in vivo models. The fellow will have many opportunities for professional development through mentoring, networking, attending conferences, and experience with grant writing. The fellow will train in the Biological Design Center at Boston University, which holds extensive expertise in molecular, cellular, and tissue engineering and presents an interdisciplinary and collaborative environment for the fellow to develop scientifically and professionally.

项目概要/摘要 心血管疾病影响着全球数百万患者，占全球死亡人数的近三分之一。 缺血或血液供应减少发生在许多心血管疾病中，是一个紧迫的健康挑战。 虽然目前的治疗主要集中于现有血管的血运重建，但一个重要的亚 由于现有的合并症，患者群体无法耐受相关的手术程序。 因此，人们对开发治疗性血管生成策略非常感兴趣，该策略旨在刺激新的血管生成。 缺血部位的血管化。虽然许多用于治疗性血管生成的基因和细胞疗法已被 经过临床试验测试，对患者的明显益处仍有待观察。迄今为止，大多数基因疗法都提供一种 或两个基因到达缺血部位，而细胞疗法则递送祖细胞或干细胞以产生旁分泌因子 并自组织成脉管系统。这些疗法的一个主要限制是无法控制 表达基因或分泌因子的时间呈现。血管生成是一个复杂且 时间调节过程，其中血管生成因子首先启动原始血管的形成 成熟因子之前的网络促进壁细胞募集和血管稳定。在学习与成长的同时 因素表明，血管生成因子和成熟因子的顺序递送有利于建立 功能性脉管系统，血管生成到成熟转变的时间如何影响功能性脉管系统 已建立的脉管系统未知。组织如何自然地感知血管生成的正确时机 成熟转变也不清楚，但结合传感器来调节血管生成和血管生成的表达 成熟基因将有利于创建剂量受控和脱靶最小化的基因疗法 影响。在该提案中，合成生物学工具将与血管化和工程模型相结合 后肢缺血的体内模型，用于评估血管生成和成熟基因表达的时间 影响功能性血管网络的形成和缺血的恢复。在目标 1 中，双通道遗传 开关将用于建立血管生成到成熟转变的时间与 血管网络功能。在目标 2 中，缺氧反应元件将用于产生缺氧调节的 控制血管生成和成熟基因诱导的基因开关。基因开关将被评估 因其能够在体内后肢缺血模型中挽救灌注。相关培训计划将制定 通过使研究员获得合成生物学和体内的新技能来获得学术生涯 模型。该研究员将通过指导、网络、 参加会议，以及撰写资助金的经验。该研究员将在生物设计中心接受培训 波士顿大学在分子、细胞和组织工程方面拥有广泛的专业知识，并提出了 为研究员提供科学和专业发展的跨学科和协作环境。