Genetic-engineered control of the immunogeneic state of vascular composite allografts during preservation

血管复合同种异体移植物保存过程中免疫原性状态的基因工程控制

• 批准号: 10435519
• 负责人: Curtis Lisante Cetrulo
• 金额: $ 45.91万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 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 2025-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10435519
• 关键词: Acute; Allogenic; Allografting; Animals; Anti-Inflammatory Agents; Attenuated; Automobile Driving; Biological Markers; Biosensing Techniques; Biosensor; Blood; Blood Vessels; Bone Marrow; Burn injury; CD34 gene; CMV promoter; Calculi; Caring; Cell Density; Cell Transplantation; Cells; Characteristics; Chimerism; Clinical; Complex; Coupled; Defect; Development; Diagnosis; Diagnostic; Dose; Engineering; Engraftment; Ensure; Ethics; Generations; Genes; Genetic; Genetic Engineering; Goals; Graft Rejection; Hand; Health; Immunosuppression; In Vitro; Inbreeding; Interleukins; Intervention; Isolated limb perfusion; Lead; Libraries; Long-Term Effects; Luciferases; Measures; Methods; Modeling; Modification; Monitor; NF-kappa B; Organ Transplantation; Patients; Performance; Perfusion; Pharmaceutical Preparations; Plasma; Proteins; Protocols documentation; Quality Control; Quality of life; Rattus; Recovery; Reporter; Response Elements; Rodent Model; Sampling; Seeds; Serum; Signal Transduction; Site; Solid; System; Technology; Temperature; Testing; Therapeutic; Time; Tissues; Transcriptional Regulation; Transplant Recipients; Transplantation; Transportation; Trauma; Upper Extremity; Vascular Graft; Vascularized Composite Allotransplantation; Work; allograft rejection; base; blood-based biomarker; cell type; clinical practice; clinically relevant; cost; dosage; efficacy testing; experimental study; facial transplantation; genetically modified cells; immunogenicity; immunoreaction; immunosuppressed; improved; in vivo; in vivo evaluation; limb transplantation; macrophage; novel; preservation; pressure; promoter; prospective; reconstruction; release factor; response; response biomarker; sensor; side effect; synthetic biology; theranostics; therapeutic protein; transcription factor

ABSTRACT While being an increasingly used option of reconstruction for severe defects arising from trauma and/or burn injuries, VCA remains limited in use due to the complex ethical calculus of a treatment that improves life quality dramatically, but at the cost of the side effects of long-term immunosuppression. In the longer term the most promising approach is the induction of tolerance by mixed chimerism by bone-marrow co-transplantation. In the shorter term, new strategies for monitoring immunosuppression to enable timely interventions, and reducing immunogenicity of VCAs are necessary for making this treatment more accessible to patients in need. The objective of this application is to develop a functional preservation platform that enables creating engineered VCA grafts by exogenous administration of genetically-modified cells, their preservation in a clinically practicable protocol, and testing the efficacy in reducing immunogenicity in rodent models of rejection and tolerance induction. Biosensor cells will be genetically engineered with a transcription factor response element as a gene promoter to serve as a theranostic, simultaneously driving the secretion of blood-based biomarker and a therapeutic protein to attenuate a rejection response. We will combine this with our perfusion-based supercooled preservation technologies to enable both engraftment of cells and to provide a time-window that makes the utilization of engineered grafts viable in clinical time frames. This objective has been formulated based on our prior work and preliminary results, where we have shown successful engineering of cells with required response characteristics, development of an ex vivo rat limb perfusion system that can serve as a platform for engrafting these cell biosensors into vascular grafts prior to transplant, and demonstrating that transducted cells can remain viable in vivo for up to 4 months. The proposed work would lead to a smart-graft technology that can sensitively measure local tissue signaling would enable practical monitoring and diagnosis of acute rejection episodes, can transform care of transplant patients since compliance issues in medication would be dramatically reduced, and will likely increase the overall efficacy and therefore reduce graft rejections. In the longer term, the preservation methods developed here could also be enabling for tolerance induction in VCA transplants, and therefore allow for their wide-spread use which is not possible currently.

抽象的 同时越来越多地用于修复因创伤和/或烧伤引起的严重缺陷 由于改善生活质量的治疗存在复杂的伦理考量，VCA 的使用仍然受到限制 显着，但代价是长期免疫抑制的副作用。从长远来看最 有前途的方法是通过骨髓共移植通过混合嵌合诱导耐受。在 短期内，监测免疫抑制的新策略能够及时干预，并减少 VCA 的免疫原性对于使有需要的患者更容易获得这种治疗是必要的。这 该应用程序的目标是开发一个功能保存平台，能够创建工程化的 通过外源施用转基因细胞进行 VCA 移植，并将其保存在临床可行的环境中 方案，并测试在啮齿动物排斥和耐受模型中降低免疫原性的功效 就职。生物传感器细胞将采用转录因子反应元件作为基因进行基因工程 启动子作为治疗诊断，同时驱动基于血液的生物标志物和 治疗性蛋白质可减弱排斥反应。我们将把它与基于灌注的过冷技术结合起来 保存技术既能实现细胞的移植，又能提供一个时间窗口，使 利用在临床时间范围内可行的工程移植物。这一目标是根据我们的 先前的工作和初步结果，我们已经展示了具有所需响应的成功的细胞工程 特点，开发可作为移植平台的离体大鼠肢体灌注系统 在移植前将这些细胞生物传感器植入血管移植物中，并证明转导的细胞可以保留 在体内可存活长达4个月。拟议的工作将导致一种智能移植技术，可以敏感地 测量局部组织信号传导将使急性排斥反应的实际监测和诊断成为可能 改变移植患者的护理，因为药物依从性问题将大大减少，并且 可能会提高总体疗效，从而减少移植物排斥反应。从长远来看，保存 这里开发的方法也可以在 VCA 移植中诱导耐受，因此允许 其广泛使用目前是不可能的。