Identification of Biomechanically Valid Cell Therapeutics for Vocal Fold Scar

描述（由申请人提供）：对于声带瘢痕患者，目前还没有有效的治疗方案来改善其声带功能和生活质量。细胞疗法被认为具有促进声带瘢痕修复的先天能力，但缺乏关于最佳细胞来源的知识阻碍了这种治疗方法的发展。候选细胞来源包括骨髓间充质基质细胞（BM-MSC）或脂肪组织间充质基质细胞（AT-MSC）。这两种细胞类型都表现出对其生物力学环境的敏感性，这可以改变细胞功能的高度相关方面，包括营养因子的分泌、细胞外基质（ECM）重塑和细胞分化倾向。迄今为止，msc介导的声带瘢痕修复的试验已经在动物模型中进行，这些模型不能在生物力学上转化为人类喉部。本应用程序的目的是确定MSC （BM和/或AT）是否是生物力学上有效的声带组织工程细胞来源。根据我们的初步数据，我们的中心假设是MSC （AT和BM）可以通过模仿人类发音的振动菌株激活为声带成纤维细胞（VFF）基因型。一个能够在体外复制声带机械环境的生物反应器将用于测试目标1和目标2的假设。特异性目的1利用DNA微阵列在人体生理水平上对比振动和拉伸应变后MSC和VFF的基因组表达。全球基因表达谱将提供生物力学基因型，作为确定MSC （BM和AT）与VFF相比作为声带固有层再生细胞来源的适用性的标准。我们期望揭示涉及ECM重塑、细胞增殖、细胞-基质和细胞-细胞粘附以及免疫调节的基因的生物力学联系。在进行临床试验之前，还必须验证长期暴露在声带生物力学力下的间充质干细胞不会引发转化为有害的间充质基质细胞衍生物，如成骨细胞、软骨细胞或成肌细胞。在具体目标2中，我们将研究机械介导的MSC分化作为时间的函数。这项工作是非常重要和创新的，因为它利用生物力学联系来回答关于理想细胞类型来源的声带再生的基本问题。此外，我们的实验设计将确定与振动诱导的VFF调节有关的基因，这很重要，因为几乎没有人知道振动应变如何改变喉部细胞。这些基因标记将为声带生物学中无数领域的研究提供信息（如声部创伤性疾病的发展和治疗），并将启动申请人的独立研究方向。

DESCRIPTION (provided by applicant): Effective treatment options are not available for improving vocal function and quality of life for patients with vocal fold scarring. Cell-based therapies are suggested to have the innate capacity for promoting vocal fold scar repair, but lack of knowledge regarding the optimal cell source has stymied the development of such therapeutics. Candidate cell sources include mesenchymal stromal cells derived from either bone marrow (BM-MSC) or adipose tissue (AT-MSC). Both cell types demonstrate sensitivity to their biomechanical environment which can alter highly relevant aspects of cell function including secretion of trophic factors, extracellular matrix (ECM) remodeling, and propensity for cell differentiation. To date, trials of MSC-mediated repair of vocal fold scar have been performed in animal models which are not biomechanically translatable to the human larynx. The objective of this application is to determine if MSC (BM and/or AT) are biomechanically valid cell sources for vocal fold tissue engineering. Based on our preliminary data, our central hypothesis is that MSC (AT and BM) can be activated to a vocal fold fibroblast (VFF) genotype by vibratory strain which mimics human phonation. A bioreactor capable of reproducing the vocal fold mechanoenvironment ex vivo will be used to test this hypothesis for Aims 1 and 2. Specific Aim 1 contrasts genomic expression of MSC and VFF following vibration and tensile strain (stretch) at human physiological levels using DNA microarray. Global gene expression profiles will provide biomechanical genotypes which will be utilized as criteria for determining suitability of MSC (BM and AT) compared to VFF as cell sources for vocal fold lamina propria regeneration. We expect to uncover biomechanical linkage for genes involving ECM remodeling, cell proliferation, cell-matrix and cell-cell adhesion, and immunomodulation. Prior to proceeding to a clinical trial, it is also essential to verify that MSC exposed to biomechanical forces of the vocal folds long-term will not trigger transformation to harmful mesenchymal stromal cell derivatives, such as osteoblasts, chondroctyes or myoblasts. In Specific Aim 2, we will examine mechanically mediated MSC differentiation as a function of time. This work is highly significant and innovative because it exploits biomechanical linkage to answer the fundamental question concerning the ideal cell type(s) to source for vocal fold regeneration. Additionally, our experimental design will identify genes implicated in vibration-induced modulation of VFF, which is important because virtually nothing is known about how vibratory strain alters the cells in the larynx. These gene markers will inform innumerable areas of inquiry in vocal fold biology (such as phonotraumatic disease development and treatment) and will also launch the applicant's independent line of research. PUBLIC HEALTH RELEVANCE: The goal of this research is to identify optimal cell source(s) for vocal fold scarring therapeutics. To this end, we will evaluate mesenchymal stromal cells (MSC) in a context which simulates the biomechanical forces of the human larynx. Study findings will advance knowledge of cell response to phonation-like vibration and inform the design of biomechanically relevant treatment for vocal fold regeneration prior to a clinical trial.