Role of Stem Cells in Skeletal Aging

干细胞在骨骼衰老中的作用

• 批准号: 7904240
• 负责人: JULIANNE GLOWACKI
• 金额: $ 17.09万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-08-01 至 2011-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7904240
• 关键词: Adult; Age; Aging; Antioxidants; Area; Biology; Bone Marrow; Bone Regeneration; Cells; Control Groups; DNA; Defect; Dose; Elderly man; Engraftment; Evaluation; Experimental Designs; Female; Femur; Fluorescent in Situ Hybridization; Grant; Healed; Hindlimb; Hour; Human; Immunohistochemistry; In Situ; In Situ Hybridization; In Vitro; Injection of therapeutic agent; Joints; Kinetics; Left; Leg; Limb structure; Marrow; Measures; Mesenchymal Stem Cells; Methods; Mitochondria; Modeling; Mus; NOD/SCID mouse; Operative Surgical Procedures; Orthopedic Surgery procedures; Orthopedics; Osteoblasts; Osteogenesis; Paraffin Embedding; Pathology; Peer Review; Premature aging syndrome; Procedures; Process; Production; Publications; Radiation-Protective Agents; Radiobiology; Recording of previous events; Relative (related person); Research; Role; Stem cells; Stromal Cells; Techniques; Testing; Tissues; Whole-Body Irradiation; Wound Healing; Y Chromosome; adult stem cell; age effect; age related; aged; bone; bone healing; design; efficacy testing; experience; healing; human subject; immunocytochemistry; in vivo; intraperitoneal; irradiation; male; men; mouse model; normal aging; novel; older men; osteoblast differentiation; progenitor; public health relevance; research study; skeletal; stem cell therapy; tibia; tool; wound

DESCRIPTION (provided by applicant): This application responds to PA-06-137 "Testing stem cell therapy in mouse models of premature aging"; this project explores in the R21 format the potential use of stem cell therapy in basic aging research. Two models of aging stem cells, one mouse and one human; will be used as donor cells for evaluation of their relative roles in bone wound healing. The SAMP6 mouse is known to be osteopenic and have fewer osteoblast progenitors in marrow compared with control SAMR1 mice. Marrow stromal cells (MSCs, a.k.a. mesenchymal stem cells), which contain osteoblast progenitors, will be prepared from male SAMP6 and SAMR1 mice and from marrow discarded from young and older men during orthopedic surgery. It is known that in vitro osteoblast differentiation of human MSCs declines with age of the subject. These murine and human MSCs will be engrafted into immunoprivileged female NOD/SCID recipient mice with a validated technique that results in greater engraftment of stromal cells because of the clearing of the recipient marrow niche with high dose irradiation only to the hindlimb. Following irradiation and before injection of MSCs, standardized unicortical tibial wounds will be made. Thus the hindlimb will contain donor stromal cells from young or old male mice or humans, allowing for tracking of their relative functional contribution to bone wound healing. In addition, we shall test the efficacy of a novel mitochondria-targeted antioxidant, JP4-039, to increase the functional contribution of the engrafted MSCs to bone wound healing. Methods include isolation of MSCs from male SAMP6 and SAMR1 mice and from young and older men undergoing orthopedic surgery, production of chimeric mice, measuring kinetics of tibial wound healing by weekly radiographs and <CT, measuring bone repair by histomorphometric evaluation, and quantifying the contribution of donor male cells to wound neo-osteogenesis by immunohistological and in situ analysis of Y- chromosome. This project is designed to reveal the degree to which aging compromises adult stem cell pools and whether a novel mitochondrial-targeted agent, JP4-039, effectively "rejuvenates" the contribution of aged MSCs, either from SAMP6 mice or from older human subjects, to enhanced bone wound healing. It will also reveal whether murine SAM cells show the same differences with age as do human cells. PUBLIC HEALTH RELEVANCE: New tools are needed to assess the effects of age on stem cells, in the contexts of normal aging and of potential stem cell therapy. This project is designed to reveal the effect of aging on marrow stromal cells' contribution to bone healing and the efficacy of a novel antioxidant on this process.

描述（由申请人提供）：本申请响应 PA-06-137“在过早衰老的小鼠模型中测试干细胞疗法”；该项目以 R21 形式探索干细胞疗法在基础衰老研究中的潜在用途。两种衰老干细胞模型，一种是小鼠，一种是人类；将用作供体细胞来评估它们在骨伤口愈合中的相对作用。已知 SAMP6 小鼠骨质疏松，与对照 SAMR1 小鼠相比，骨髓中的成骨细胞祖细胞较少。骨髓基质细胞（MSC，又名间充质干细胞）含有成骨细胞祖细胞，将从雄性 SAMP6 和 SAMR1 小鼠以及骨科手术期间年轻和老年男性丢弃的骨髓中制备。已知人MSC的体外成骨细胞分化随着受试者的年龄而下降。这些小鼠和人类 MSC 将被移植到免疫豁免的雌性 NOD/SCID 受体小鼠中，采用经过验证的技术，由于仅对后肢进行高剂量照射，可清除受体骨髓生态位，从而导致基质细胞更大的植入。照射后和注射间充质干细胞之前，将制作标准化的单皮质胫骨伤口。因此，后肢将含有来自年轻或年老雄性小鼠或人类的供体基质细胞，从而可以追踪它们对骨伤口愈合的相对功能贡献。此外，我们将测试一种新型线粒体靶向抗氧化剂 JP4-039 的功效，以增加植入的 MSC 对骨伤口愈合的功能贡献。方法包括从雄性 SAMP6 和 SAMR1 小鼠以及接受骨科手术的年轻和老年男性中分离 MSC，产生嵌合小鼠，通过每周 X 光照片和 CT 测量胫骨伤口愈合动力学，通过组织形态学评估测量骨修复，并通过免疫组织学和 Y-原位分析量化供体雄性细胞对伤口新骨生成的贡献。 染色体。该项目旨在揭示衰老对成体干细胞库的损害程度，以及新型线粒体靶向剂 JP4-039 是否能够有效地“恢复”衰老 MSC（无论是来自 SAMP6 小鼠还是来自老年人类受试者）对增强骨伤口愈合的贡献。它还将揭示小鼠 SAM 细胞是否表现出与人类细胞相同的随年龄变化的差异。公共健康相关性：在正常衰老和潜在干细胞治疗的背景下，需要新的工具来评估年龄对干细胞的影响。该项目旨在揭示衰老对骨髓基质细胞对骨愈合的影响，以及新型抗氧化剂对此过程的功效。