Novel Strategies in Bladder Tissue Engineering: Stem Cells and Silk Biomaterials

项目摘要/摘要 我在这项提案中的总体目标是解决目前膀胱手术选择上的局限性。 重建，同时发展我的专业能力，过渡到一名独立的调查员。我的 职业目标是在一所一流的学术机构担任实验室主任，这将使我 通过创造基于干细胞的创新疗法，继续拓展再生医学的前沿 用于泌尿系统组织重建。我的主要研究培训目标将是获得新的技术 泌尿外科组织工程系统和人类多能干细胞生物学方面的技能。这将是 通过额外的基础课程和实验室方案的实践指导来完成 如提案目标中详细说明的技术。职业发展机制将包括具有启发性的 资助撰写、学术工作面试、实验室管理等方面的研讨会和指导 手稿准备工作。一个咨询委员会将评估我的科学和职业生涯的完成情况 发展里程碑，并促进我向独立调查员的转变。 膀胱的组织工程学方法最近被开发出来，利用 种植自体平滑肌细胞(SMC)的3D支架。这些策略导致了部分 短期临床试验中器官功能的恢复。然而，这些构造对于 完整的缺陷修复可能受到传统生物材料载体的次优性能的限制 功能上有能力的细胞来源的可用性。基于丝绸的生物材料提供了一种特殊的 非常适合支持膀胱功能的身体特征的组合，因此可以 解决与当前支架材料相关的限制。多能干细胞群体，如 胚胎干细胞(ESCs)和/或诱导的多能干细胞(IPS)可以满足以下需求 能够产生器官和患者特定的SMC的替代细胞来源。为了解决 目前膀胱功能组织工程(FTE)的局限性，我们将挑战总体假设： 使用丝质生物材料种植原代或干细胞来源的SMC的FTE模式将促进 平滑肌缺损巩固和膀胱功能恢复。这项研究的具体目的 目的1：确定原代SMC接种丝质生物材料对膀胱的治疗效果 平滑肌组织再生。目的2：评价ESC和iPS来源的SMC作为细胞的实用性 功能性膀胱组织工程的来源。

PROJECT SUMMARY/ABSTRACT My overall goal in this proposal is to address the current limitations in surgical options for bladder reconstruction while developing my professional abilities to transition into an independent investigator. My professional objective is to have a position as a lab director in a superb academic institution that will allow me to continue to expand the frontiers of regenerative medicine by creating innovative stem cell-based therapies for urological tissue reconstruction. My primary research training objectives will be to acquire new technical skills in urological tissue engineered systems and human pluripotent stem cell biology. This will be accomplished through additional fundamental coursework and hands on instruction in laboratory protocols and techniques as detailed in the aims of the proposal. Career development mechanisms will include instructive seminars and mentored guidance in grant writing, academic job interviewing, laboratory management, and manuscript preparation. An advisory committee will evaluate the completion of both my scientific and career development milestones and facilitate my transition into an independent investigator. Tissue engineering approaches for the urinary bladder have recently been developed utilizing 3D scaffolds seeded with autologous smooth muscle cells (SMCs). These strategies have led to partial restoration of organ functionality in short-term clinical trials. However, the efficacy of these constructs for complete defect repair may be limited by the suboptimal properties of conventional biomaterial carriers and availability of functionally competent cell sources. Silk-based biomaterials provide an exceptional combination of physical characteristics that are well suited to support bladder function and therefore may address limitations associated with current scaffold materials. Pluripotent stem cell populations such as embryonic stem cells (ESCs) and/or induced pluripotent stem (iPS) cells may fulfill the need for alternative cell sources capable of generating organ and patient-specific SMCs. In order to address the current limitations in bladder functional tissue engineering (FTE), we will challenge the overall hypothesis that: FTE modalities using silk-based biomaterials seeded with primary or stem cell-derived SMCs will promote smooth muscle defect consolidation and restoration of bladder function. The specific aims of the study are: Aim 1: Determine the efficacy of silk-based biomaterials seeded with primary SMCs for bladder smooth muscle tissue regeneration. Aim 2: Evaluate the utility of ESC and iPS-derived SMCs as cell sources for functional bladder tissue engineering.