Identification of Biomechanically Valid Cell Therapeutics for Vocal Fold Scar

描述（由申请方提供）：目前尚无有效的治疗方案可用于改善声带瘢痕患者的发声功能和生活质量。基于细胞的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疗法的发展。候选细胞来源包括来源于骨髓（BM-MSC）或脂肪组织（AT-MSC）的间充质基质细胞。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，这些动物模型在生物力学上不能转化为人类喉部。本申请的目的是确定MSC（BM和/或AT）是否是用于声带组织工程的生物力学有效的细胞来源。基于我们的初步数据，我们的中心假设是MSC（AT和BM）可以通过模拟人类发声的振动应变激活为声带成纤维细胞（VFF）基因型。将使用能够离体再现声带机械环境的生物反应器来检验目的1和2的这一假设。具体目标1对比基因组表达的MSC和VFF后振动和拉伸应变（拉伸）在人类生理水平使用DNA微阵列。全局基因表达谱将提供生物力学基因型，其将用作确定MSC（BM和AT）与VFF相比作为声带固有层再生的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于声带生物力学力的MSC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声带再生的理想细胞类型的基本问题。此外，我们的实验设计将确定与振动诱导的VFF调制有关的基因，这很重要，因为几乎不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（如声音创伤性疾病的发展和治疗）的无数领域提供信息，并将启动申请人的独立研究。

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.