Noninvasive preconditioning of mesenchymal stem cells to improve potency for bone repair

无创预处理间充质干细胞以提高骨修复效力

• 批准号: 10940869
• 负责人: Rebekah Margaret Samsonraj
• 金额: $ 8.85万
• 依托单位: UNIV OF ARKANSAS FOR MED SCIS
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-10-03 至 2028-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10940869
• 关键词: Allogenic; Biomechanics; Biophysics; Bone Regeneration; Calcium Channel; Calvaria; Cell Aging; Cell Therapy; Cell surface; Cells; Cephalic; Characteristics; Cues; Dasatinib; Defect; Exposure to; Gene Cluster; Gene Expression; Gene Expression Profiling; Genomic approach; Growth Factor; Heterogeneity; Human; In Vitro; Mechanical Stimulation; Mesenchymal Stem Cells; Modeling; Mus; Musculoskeletal Diseases; Osteoblasts; Osteogenesis; Outcome; Pathway interactions; Phenotype; Piezo 1 ion channel; Proliferating; Quercetin; Regenerative Medicine; Research; Safety; Serial Passage; Signal Transduction; Surface; Tissue-Specific Gene Expression; Transplantation; Transplantation Conditioning; bone; bone healing; bone repair; cell preparation; chemokine; cytokine; genetic manipulation; healing; histological stains; immunoregulation; improved; in vivo; innovation; irradiation; mechanical signal; novel; osteoblast differentiation; osteogenic; preconditioning; regenerative therapy; response; senescence; single-cell RNA sequencing; small molecule; stem cell population; stem cells; therapy outcome; tissue repair; vibration

Mesenchymal stem cells (MSCs) are trialed as therapies in bone regenerative medicine owing to their ability to differentiate into osteogenic, chondrogenic and myogenic lineages. However, therapeutic outcomes vary greatly. The basis for this inconsistency is unclear but can be attributed to MSC heterogeneity characterized by donor-to-donor variability and intra-donor heterogeneity in phenotypic and functional characteristics. Unfortunately, current strategies to improve MSC potency for bone repair via preconditioning with exogenous growth factors or genetic manipulation have limited potential for FDA approval owing to efficacy and safety concerns. Here, we propose novel noninvasive approaches to improve MSC functionality using biophysical stimulation and senescence clearance in heterogenous cultures. The objective of this project is to determine whether MSC potency for bone repair can be improved by (i) non-invasive, biomechanical signals and (ii) modulating cellular senescence in heterogeneous MSC populations. We propose three aims: (i) determine if non-invasive mechanical signaling promotes osteoblastic differentiation of MSCs and their bone-repair potency. (ii) determine the impact of MSC senescence on bone-repair potency and, (iii) define MSC heterogeneity using single-cell RNA sequencing and correlate key heterogeneity parameters with in vivo outcomes of new bone formation. To execute these aims, MSCs will be delivered low magnitude vibrations (LMV) and assessed for in vitro osteogenic differentiation and in vivo bone formation within critical-sized calvarial defect in mice. Secondly, heterogenous cultures of MSCs characterized by significant presence of senescent cells and associated secretory phenotype will be treated with senolytics followed by assessment for osteogenic differentiation capacity, cytokine and trophic factor secretion, and in vivo bone healing in a calvarial defect model. Thirdly, we will investigate gene expression differences related to key MSC characteristics such as proliferation, differentiation, and cell surface phenotype using single cell genomics approach on multiple donor MSCs. Unraveling stem cell subpopulations that may determine functional potency will be critical for the selection of suitable donors for allogeneic cell therapies. The proposed innovative use of LMV and senolytics as pre-transplantation conditioning to achieve homogeneous and potent MSC preparations holds significance in developing successful cell-based regenerative therapies for cranial tissue repair.

间充质干细胞（MSC）由于能够分化成成骨、软骨和肌源谱系，因此被尝试作为骨再生医学的疗法。然而，治疗结果差异很大。这种不一致的原因尚不清楚，但可归因于 MSC 异质性，其特征是供体之间的变异性以及供体内部表型和功能特征的异质性。不幸的是，由于功效和安全性问题，目前通过外源生长因子预处理或基因操作来提高 MSC 骨修复效力的策略获得 FDA 批准的潜力有限。在这里，我们提出了利用异质培养物中的生物物理刺激和衰老清除来改善间充质干细胞功能的新型非侵入性方法。该项目的目标是确定是否可以通过 (i) 非侵入性生物力学信号和 (ii) 调节异质 MSC 群体中的细胞衰老来提高 MSC 骨修复功效。我们提出三个目标：(i) 确定非侵入性机械信号传导是否促进 MSC 的成骨细胞分化及其骨修复能力。 (ii) 确定 MSC 衰老对骨修复能力的影响，(iii) 使用单细胞 RNA 测序定义 MSC 异质性，并将关键异质性参数与新骨形成的体内结果相关联。为了实现这些目标，MSC 将被传递低强度振动 (LMV)，并评估小鼠临界尺寸颅骨缺损内的体外成骨分化和体内骨形成。其次，以明显存在衰老细胞和相关分泌表型为特征的 MSC 异质培养物将用 senolytics 处理，然后评估成骨分化能力、细胞因子和营养因子分泌以及颅骨缺损模型中的体内骨愈合。第三，我们将使用单细胞基因组学方法在多个供体 MSC 上研究与关键 MSC 特征（例如增殖、分化和细胞表面表型）相关的基因表达差异。揭示可能决定功能效力的干细胞亚群对于选择合适的供体进行同种异体细胞疗法至关重要。所提出的创新性使用 LMV 和 senolytics 作为移植前调理以实现均质和有效的 MSC 制剂对于开发成功的基于细胞的颅组织修复再生疗法具有重要意义。