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

描述(由申请人提供)：我在这个提案中的总体目标是解决目前膀胱重建手术选择上的局限性，同时发展我的专业能力，以过渡到一名独立的研究人员。我的职业目标是在一所一流的学术机构担任实验室主任的职位，这将使我能够通过为泌尿系统组织重建创造基于干细胞的创新疗法来继续扩大再生医学的前沿。我的主要研究培训目标将是获得泌尿系统组织工程系统和人类多能干细胞生物学方面的新技术技能。这将通过额外的基础课程和实验室方案和技术的实际指导来实现，如提案的目的所详述。职业发展机制将包括在补助金撰写、学术工作面试、实验室管理和手稿准备方面的指导性研讨会和指导指导。一个咨询委员会将评估我的科学和职业发展里程碑的完成情况，并促进我向独立调查员的转变。最近，利用种植有自体平滑肌细胞(SMCs)的3D支架，已经开发出用于膀胱的组织工程方法。这些策略已经在短期临床试验中部分恢复了器官功能。然而，这些构建物用于完全修复缺陷的有效性可能会受到传统生物材料载体的次优性能和功能合格细胞来源的可用性的限制。基于丝素的生物材料提供了非常适合支持膀胱功能的物理特性的特殊组合，因此可能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)，可能满足对能够产生器官和患者特异性SMC的替代细胞来源的需求。为了解决目前膀胱功能组织工程(FTE)的局限性，我们将挑战以下总体假设：以丝素为基础的生物材料种植原代或干细胞来源的SMC将促进平滑肌缺损的巩固和膀胱功能的恢复。本研究的具体目的是：目的1：确定丝质生物材料与原代SMC复合后对膀胱平滑肌组织再生的效果。目的：评价ESC和iPS来源的SMC作为功能性膀胱组织工程细胞来源的可行性。 公共卫生相关性：这项提案的结果预计将产生新的组织再生策略，将克服目前膀胱重建的生物材料和细胞来源限制。这项研究将提供比我们的具体目标范围更广的新信息，可能适用于其他泌尿生殖组织(如尿路)的修复，也可能适用于其他中空器官的修复。

DESCRIPTION (provided by applicant): 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. PUBLIC HEALTH RELEVANCE: The results of this proposal are anticipated to produce new tissue regeneration strategies that will overcome current biomaterial and cell source limitations for urinary bladder reconstruction. This study will provide new information that is wider in scope than our specific goals and which may be applicable for repair of other urogenital tissues (e.g. urethra) and possibly other hollow organs.