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 虽然是由于创伤和/或燃烧引起的严重缺陷的重建越来越多的选择 受伤，VCA由于修复的复杂伦理演算，可改善生命质量 急剧，但要付出长期免疫抑制的副作用。从长远来看 有前途的方法是通过骨髓共转移诱导混合嵌合感诱导耐受性。在 较短的任期，监测免疫抑制以及时干预并减少的新策略 VCA的免疫原性对于使需要的患者更容易获得这种治疗方法。这 该应用程序的目的是开发一个功能保护平台，该平台能够创建工程 通过外源给予遗传改性细胞的VCA移植物，它们在临床上可行的保存 方案，并测试降低啮齿动物排斥和公差模型中免疫原性的功效 就职。生物传感器细胞将以转录因子响应元件作为基因进行基因设计 发起人作为一种疗法，同时推动基于血液的生物标志物的分泌 治疗性蛋白质可减轻排斥反应。我们将将其与基于灌注的超冷 保存技术可以使细胞植入植入，并提供一个时间窗口，使得 在临床时间范围内可行的工程移植物的利用。这个目标是根据我们的 先前的工作和初步结果，我们已经显示了细胞的成功工程，并需要响应 特征，离体大鼠肢体灌注系统的开发，可以用作雕刻的平台 这些细胞生物传感器在移植前进入血管移植物，并证明转导细胞可以保留 在体内可行长达4个月。拟议的工作将导致一种智能的技术，可以敏感 衡量局部组织信号传导将使急性排斥发作的实际监测和诊断，可以 转变移植患者的护理，因为药物方面的依从性问题将大大减少，并且 可能会提高整体功效，从而减少移植物拒绝。从长远来看，保存 此处开发的方法也可以使VCA移植的耐受性诱导，因此允许 对于他们的广泛使用，目前是不可能的。