Engineered Asymmetric Hydrogel for Muscle Stem Cell Polarity and Fate Specification

用于肌肉干细胞极性和命运规范的工程不对称水凝胶

• 批准号: 10405716
• 负责人: Woojin Han
• 金额: $ 45.54万
• 依托单位: ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-03-01 至 2027-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10405716
• 关键词: Address; Adhesives; Apical; Basal lamina; Biochemical; Biocompatible Materials; Biological Assay; Biomechanics; Biophysics; Cachexia; Cell Culture System; Cell Polarity; Cell Survival; Cell Therapy; Cell division; Cells; Centrosome; Cues; Disease; Dystrophin; Elasticity; Employment; Engineering; Engraftment; Epidermal Growth Factor Receptor; Equilibrium; Genes; Hydrogels; In Vitro; Instruction; Integrins; Life Cycle Stages; Ligands; Maintenance; Mechanics; Mitotic spindle; Muscle; Muscle Fibers; Muscle satellite cell; Muscular Dystrophies; Myogenin; Outcomes Research; Patients; Peptides; Periodicity; Physiological; Proteins; Regenerative Medicine; Regulation; Research; Role; Signal Transduction; Skeletal Muscle; Source; Stem cell transplant; Stretching; System; Testing; Therapeutic; Transplantation; Trauma; adult stem cell; aging population; aurora kinase A; base; beta catenin; cell type; cellular engineering; combinatorial; design; ethylene glycol; mechanical force; mechanical stimulus; mimetics; muscle regeneration; planar cell polarity; receptor; sarcopenia; satellite cell; segregation; self-renewal; socioeconomics; stem cell biology; stem cell division; stem cell expansion; stem cell niche; stem cells; stemness; volumetric muscle loss

ABSTRACT Skeletal muscle stem cell (MuSC) transplantation is emerging as a promising strategy for treating muscle- associated trauma and diseases. However, MuSCs spontaneously lose their “stemness” and engraftment potential in conventional 2D culture, critically limiting their applicability for cell-based therapy. This inability to culture MuSCs further limits the employment of cell engineering strategies, such as ex vivo gene editing patient derived cells. Thus, there is a critical need for a strategy to maintain and expand therapeutically potent MuSC ex vivo long-term. To date, MuSC culture systems that recapitulate the asymmetric MuSC niche, consisting of myofiber and basal lamina, required to establish cellular polarity, support physiologic cell division, and guide fate specification, do not exist. The contribution of active mechanical forces in modulating MuSC polarity, division, and fate specification also remains overlooked. We believe that establishing MuSC polarity is a requirement for supporting long-term symmetric expansion ex vivo through biochemical and biophysical means. To this end, the proposed research will engineer an asymmetric designer platform for expanding self-renewing MuSCs ex vivo long-term by directing cellular polarity and fate specification. In Aim 1, we will identify minimally essential asymmetric cues for establishing MuSC polarity by engineering an asymmetric hydrogel. In Aim 2, we will determine the effects of biochemically promoting symmetric cell division on long-term expansion of MuSCs using the asymmetric hydrogel. Biochemical strategies to inhibit asymmetric division and to promote symmetric division to achieve ex vivo MuSC expansion will be tested. In Aim 3, we will determine the effects of dynamic biomechanical stretch on regulation of symmetric MuSC expansion using the asymmetric hydrogel. Mechanisms by which externally applied forces align mitotic spindle orientation and regulate cell division of polarized MuSCs will be tested. Successful outcomes of this research will not only develop a platform for expanding MuSCs for cell therapies by directing stem cell polarity and fate specification but also provide establish a new paradigm for harnessing stem cell polarity in cell-instructive biomaterials for regenerative medicine.

抽象的 骨骼肌干细胞（MUSC）移植是作为治疗肌肉的有前途的策略 相关的创伤和疾病。但是，MUSC的赞助会失去其“茎”和植入 在常规2D培养物中的潜力，严重限制了它们对基于细胞的治疗的适用性。这无力 培养物进一步限制了细胞工程策略的使用，例如离体基因编辑患者 衍生细胞。这是迫切需要一项策略来维护和扩展治疗潜在的MUSC 长期体内。迄今 肌纤维和碱性薄片，需要建立细胞极性，支持生理细胞分裂和指导 命运规范，不存在。主动机械力在调节MUSC极性时的贡献， 部门和命运规范也被忽略了。我们认为建立MUSC极性是 通过生化和生物物理手段支持长期对称扩展的要求。 为此，拟议的研究将设计一个不对称的设计师平台，用于扩大自我更新 MUSC长期通过指导细胞极性和命运规范。在AIM 1中，我们将最少识别 通过工程不对称水凝胶来建立MUSC极性的必需不对称提示。在AIM 2中，我们 将确定生化促进对称细胞分裂对MUSC长期扩张的影响 使用不对称水凝胶。抑制不对称分裂并促进对称的生化策略 将测试实现离体MUSC扩展的划分。在AIM 3中，我们将确定动态的影响 使用不对称水凝胶调节对称MUSC扩展的生物力学拉伸。 外部应用力对齐有丝分裂纺锤体方向并调节细胞分裂的机制 两极化的MUSC将进行测试。这项研究的成功结果不仅会为 通过指导干细胞极性和命运规范来扩展细胞疗法的MUSC，但也提供 建立一个新的范式，用于利用细胞结构生物材料中的干细胞极性来再生 药品。