Autologous cell engineered bladder augmentation

在英国，大约有300万人经历过尿失禁，NHS每年的管理费用超过5亿英镑。它可以由先天性异常引起，如胎儿膀胱发育不全（膀胱外溢），也可以由后天条件引起，如膀胱的顺应性丧失（神经性膀胱）或膀胱癌。对于终末期膀胱疾病，最常用的增加或替换膀胱的外科手术包括纳入一个血管化的、重新配置的肠段（肠囊成形术）。虽然这种手术可以控制尿失禁并提高生活质量，但它通常与严重的并发症有关，包括感染、粘液产生和结石形成，以及长期的癌症风险。这些并发症主要是由于肠黏膜在结构和生理上与长期暴露于尿液不相容。已经提出了许多膀胱工程策略，这些策略通常依赖于将天然或合成的生物材料结合到膀胱中，这些材料可能会或不会与来自患者（自体）的细胞预培养。尽管有一些高调的报道，但很少有证据表明膀胱组织工程可以达到与当前外科技术相当的功能结果，并且仍然存在未满足的临床需求。该项目的目的是通过自体细胞工程策略为患者提供功能性膀胱重建。该项目将汇集干细胞和泌尿学（ICH， GOSH和UCLH）与尿路上皮细胞生物学和膀胱组织工程（约克大学）的专业知识，旨在开发一种膀胱重建的新技术，该技术将首先适用于膀胱外翻患者，但最终将扩展到所有需要膀胱增强的患者。该项目的阶段将首先开发方法，以确定和分离膀胱外翻病例中最合适的自体上皮细胞来源。为了达到这一目的，将从正常和病变膀胱以及口腔粘膜活检中分离和鉴定细胞。上皮细胞的扩增将使用“良好生产规范”技术进行，这是一种用于制药产品的标准方法，为转化为临床提供了一条现成的途径。人类细胞将异种移植到24头猪身上，用于膀胱增强，并在移植后3个月和6个月获得功能和组织学数据。主要结局指标将是：以生存期衡量的安全性；通过新膀胱的容量和顺应性来衡量疗效。这一阶段的次要措施将包括：手术发病率，增强和原生粘膜的死后免疫组织学分析，以及“反向翻译”结果措施，其中观察细胞行为用于在体内这些环境中提出关于干细胞/细胞功能植入的假设。

Some 3 million people in the UK experience incontinence and the cost of management is over £500 million per annum to the NHS. It can be caused by both congenital anomalies, such as a failure of the bladder in the foetus to form (bladder exstrophy), and acquired conditions such as a lost of the compliance of the bladder (neuropathic bladder) or bladder cancer. For end-stage bladder disease, the most commonly performed surgical procedure to augment or replace the bladder involves incorporating a vascularised, reconfigured segment of bowel (enterocystoplasty). Although this procedure can provide continence and enhance quality of life, it is commonly associated with serious complications, including infections, mucus production and stone formation, and long-term risk of cancer. These complications are primarily due to the fact that bowel mucosa is structurally and physiologically incompatible with the long-term exposure to urine. A number of bladder engineering strategies have been proposed and typically, these rely upon the incorporation of a natural or synthetic biomaterial into the bladder, which may or not be pre-cultured with cells derived from the patient (autologous). Despite some high profile reports, there is little evidence that bladder tissue engineering can yet achieve a functional outcome equivalent to current surgical techniques, and there remains an unmet clinical need. The aim of this programme is to deliver functional bladder reconstruction to patients through an autologous cell engineering strategy. This project will bring together expertise in stem cells and urology (ICH, GOSH and UCLH) with urothelial cell biology and bladder tissue engineering (York) with the purpose of developing a novel technique for bladder reconstruction that will be applicable in the first instance to patients with bladder exstrophy, but eventually extended to all patients requiring bladder augmentation. The stages of the project will be firstly to develop methods to identify and isolate the most appropriate source of autologous epithelial cells in the case of bladder exstrophy. To meet this aim, cells derived from normal and diseased bladder and from biopsies of buccal mucosa will be isolated, and characterized. Expansion of the epithelial cells will be conducted using "Good Manufacturing Practices" techniques, a standard methodology used for pharmaceutical products, to provide a ready route for translation to the clinic. Human cells will be xenogeneic transplanted in 24 pigs for bladder augmentation and functional and histological data will be obtained at 3 and 6 months after transplantation. Principal outcome measures will be: safety as measured by survival; efficacy as measured by capacity and compliance of the neo-bladder. Secondary measures for this phase will include: surgical morbidity, post-mortem immunohistological analysis of both the augmented and native mucosa, and also 'reverse translational' outcome measures, where observations of cellular behaviour are used to develop hypotheses about stem cell/cell functional engraftment in these settings in vivo.