Monitoring graft responses to the transplant niche in allogeneic cell replacement therapy

项目摘要/摘要 1型糖尿病患者通过每日外源性胰岛素调节血糖 给药--通往胰岛素独立之路的唯一其他选择是供体胰岛/整个器官 移植，一种有限的资源，可以取代患者由于自身免疫而丢失的细胞队列 进攻。这些移植的成功与否有很大的变数，有几名患者需要第二次或 有时需要第三次输注供体细胞。同种异体细胞/组织移植失败通常是由于免疫 宿主内的激活和强大的全身性免疫抑制是成功植入 移植手术。已经采取了许多方法来缓解对系统性的需求 免疫抑制-那些专注于操纵移植物的人已经从基因上移除了细胞的表达 表面蛋白，当不匹配时，导致适应性和先天免疫系统的激活。是这样的 细胞对免疫系统是潜在的不可见的，并构成了潜在的严重问题 感染的水库。如果我们有一种方法来识别移植物何时经历炎性应激，我们 可以设计急性抗炎疗法来保护移植物免受攻击-延长移植物的时间 移植物需要适应这个利基市场并开始运作。最近，干细胞已经 作为供体胰岛的替代品，出现了无限的细胞来源，目前正在进行早期临床尝试 正在进行中。干细胞衍生品的替代绕过了有限的移植材料的障碍-- 由于移植后的异体反应，他们仍然面临着免疫激活的问题。 我们的目标是使用我们首创的最先进的技术来应对这些挑战。结合了 利用细胞内生物传感系统提供实时反馈的干细胞治疗的潜力 应对移植利基，我们将跨越异基因细胞移植进入未来--例如 检测系统可以测量移植物的活性，可以预测移植物的成功植入并帮助 明确干预措施，以保护细胞在移植后不久免受炎性应激。我们有 结合并响应miRNA水平变化并作为细胞内传感器的工程纳米探针 为了细胞健康。我们的纳米探针是特异的、敏感的，可以在动物模型中进行活体检测。 在这里，我们测试了我们的先导炎症敏感生物传感器在干细胞来源的胰岛素产生有机化合物-这 在体外和小动物模型中，组合可以提供独特的光谱读数，这是由于 在细胞中触发的特定炎症反应，降低了我们最终的细胞治疗产品将成为 对1型糖尿病患者的功能性治疗的一部分。

Project Summary/Abstract Patients living with Type 1 Diabetes regulate their blood glucose through daily exogenous insulin administration – the only other option towards a path to insulin independence is donor islets/whole organ transplant, a limited resource, that replaces the cohort of cells that are lost in patients due to an autoimmune attack. The success of these transplants is highly variable, with several patients requiring a second or sometimes a third infusion of donor cells. Failure of allogeneic cell/tissue transplant often results from immune activation within the host, and strong systemic immunosuppression is necessary for successful engraftment of the transplant. Numerous approaches have been undertaken to alleviate the need for systemic immunosuppression – those focused on manipulating the graft have genetically removed the expression of cell surface proteins that, when mis-matched, lead to activation of the adaptive and innate immune systems. Such cells are potentially invisible to the immune system and pose the grave problem of serving as potential reservoirs of infection. If we had a method of identifying when a graft was experiencing inflammatory stress, we could design acute anti-inflammatory therapies to protect the graft from the assault – prolonging the time it takes for the graft to get accustomed to the niche and begin functioning. More recently, stem cells have emerged as a limitless resource of cells as a surrogate to donor islets, and early clinical attempts are currently underway. Replacement with stem cell derivatives circumvents the roadblock of limited transplant material – they are still faced with immune activation due to an allo-response after transplantation. Our goal is to address these challenges using state-of-the-art technologies we have pioneered. Combining the potential of stem cell therapy with an intracellular biosensing system that provides real time feedback in response to the transplantation niche, we will leapfrog allogeneic cell transplantation into the future – such a detection system can measure viability of grafts that can be predictive of successful engraftment and help define interventions to protect cells against inflammatory stress soon after transplantation. We have engineered nanoprobes that bind and respond to changes in miRNA levels and serve as intracellular sensors for cell health. Our nanoprobes are specific, sensitive, and can be detected in vivo in animal models. Here, we test our lead inflammation-sensitive biosensor in stem cell-derived insulin producing organoids – this combination can provide unique spectral read outs, in vitro and in small animal models, that result from a specific inflammatory response triggered in the cells, de-risking our final cell therapy product that will become part of a functional cure for patients living with type 1 diabetes.