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）移植正在成为治疗肌肉萎缩症的一种有前景的策略。 然而，MuSC 会自发地失去其“干性”和移植能力。 传统二维培养中的潜力，严重限制了它们在基于细胞的治疗中的适用性。 培养 MuSC 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 因此，迫切需要一种策略来维持和扩展具有治疗作用的 MuSC。 迄今为止，MuSC 培养系统概括了不对称的 MuSC 生态位，包括： 肌纤维和基底层，需要建立细胞极性、支持生理细胞分裂并引导 命运规范，不存在主动机械力在调节 MuSC 极性中的贡献。 分裂和命运规范也仍然被忽视。我们认为建立 MuSC 极性是一个问题。 通过生化和生物物理手段支持离体长期对称扩张的要求。 为此，拟议的研究将设计一个不对称的设计平台，以扩展自我更新 通过指导细胞极性和命运规范，MuSC 可以在体外长期存在。在目标 1 中，我们将最低限度地进行鉴定。 在目标 2 中，我们通过设计不对称水凝胶来建立 MuSC 极性的基本不对称线索。 将确定生化促进对称细胞分裂对 MuSC 长期扩增的影响 使用不对称水凝胶抑制不对称分裂并促进对称分裂。 在目标 3 中，我们将测试实现离体 MuSC 扩增的分裂效果。 使用不对称水凝胶调节对称 MuSC 扩张的生物力学拉伸。 外加力调整有丝分裂纺锤体方向并调节细胞分裂的机制 极化 MuSC 将接受测试，这项研究的成功成果不仅会开发一个平台。 MuSC 通过指导干细胞极性和命运规范来进行细胞治疗，还提供 建立一个新的范例，利用细胞指导生物材料中的干细胞极性进行再生 药品。