Enhanced Bone Healing Around Implants by Transplanted NF-kB Driven Immunomodulating MSCs

通过移植 NF-kB 驱动的免疫调节 MSC 增强植入物周围的骨愈合

• 批准号: 10222569
• 负责人: STUART B GOODMAN
• 金额: $ 33.52万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-09-15 至 2023-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10222569
• 关键词: Acute; Age; Aging; Animals; Anti-Inflammatory Agents; Biological Assay; Bone Density; Bone Diseases; Bone Marrow; Bone remodeling; CMV promoter; Cells; Chronic; Coculture Techniques; Custom; Disease; Elderly; Environment; Exposure to; Female; Fluorescence; Grant; Immune; Implant; Inflammation; Inflammatory; Infusion procedures; Interleukin-4; Lentivirus Vector; Lipopolysaccharides; Luciferases; Medical Device; Mesenchymal Differentiation; Mesenchymal Stem Cell Transplantation; Mesenchymal Stem Cells; Modeling; Molecular; Mus; NF-kappa B; Orthopedics; Osseointegration; Osteoclasts; Osteogenesis; Osteolysis; Outcome; Phase; Phenotype; Plasmids; Population; Process; Production; Prosthesis; Replacement Arthroplasty; Reporter; Research; Signal Transduction; Stimulus; System; TNF gene; Therapeutic Effect; Tissues; Transplantation; aged; arginase; body system; bone; bone healing; bone loss; chemokine; chronic inflammatory disease; clinically relevant; cytokine; efficacious treatment; immunoregulation; implantation; improved; in vivo; inflammatory bone loss; macrophage; male; minimally invasive; novel; osteogenic; paracrine; particle; preconditioning; regenerative; regenerative tissue; sex; stem cell therapy; stem cells; transcription factor; translational approach

Acute transient inflammation is crucial for initiation of bone healing, osseointegration of implants, osteogenic differentiation of MSCs, and immunomodulation. However, harmful chronic inflammation and bone loss (osteolysis) are induced by adverse stimuli including wear byproducts from joint replacements (JR). Nuclear Factor kappa B (NF-κB) is a critical transcription factor in both macrophages (mϕ) and MSCs that regulates inflammation, bone formation/remodeling, and aging. We showed that preconditioning MSCs with lipopoly- saccharide (LPS) and tumor necrosis factor-α (TNF-α) to induce acute transient activation of NF-κB synergistically enhances osteogenesis, and modulates mϕ polarization from a pro-inflammatory (M1) to an anti-inflammatory pro-regenerative (M2) phenotype. Furthermore, inhibition of persistent NF-κB signaling using an NF-κB decoy OligoDeoxyNucleotide (ODN) was shown to mitigate inflammatory bone loss. We have genetically modified MSCs to sense NF-κB activation and then increase production of the anti-inflammatory pro-regenerative cytokine IL-4. The purpose of this grant is to accelerate net bone formation and mitigate chronic inflammation and osteolysis via NF-kB driven immunomodulation using preconditioned or genetically modified MSCs transplanted to the local environment. Specific Aim #1a -To define the critical immunomodulatory interactions of preconditioned vs. NF-κB sensing and IL-4 secreting MSCs on mϕ exposed to wear byproducts from orthopaedic implants. Specific Aim #1b - To enhance osteogenesis by using preconditioned vs. NF-κB sensing and IL-4 secreting MSCs in an MSC/mϕ co-culture model exposed to wear byproducts from orthopaedic implants. Specific Aim #2a - To demonstrate the therapeutic effects of transplanted preconditioned MSCs on bone during the acute inflammatory stage (simulating the stage of initial implantation of a prosthesis and osseointegration) using the murine continuous femoral particle infusion model. Specific Aim #2b - To demonstrate the therapeutic effects of preconditioned vs. NF-κB sensing and IL-4 secreting MSCs on net bone formation during the chronic inflammatory stage (simulating established wear particle disease) using the murine continuous femoral particle infusion model. Specific Aim #3 - To demonstrate that the above principles of modulating acute and chronic inflammation are valid irrespective of animal sex (male vs. female mice) or age (young vs. elderly mice) in vivo. We hypothesize that NF-κB preconditioning or NF-κB sensing and IL-4 secreting MSCs will enhance the osteogenic and immunomodulatory signaling of MSCs, enhancing bone formation via crosstalk by mϕ and MSCs. NF-kB driven immunomodulation of MSCs should mitigate particle-induced inflammation, osteoclast activation, and enhance bone formation in young and aged, male and female mice. NF-kB driven immune modulation of MSCs is novel, mechanistic, and directly translational to inflammatory disorders in other organ systems.

急性瞬态感染对于骨骼愈合的启动至关重要 MSC的分化和免疫调节。但是，有害的慢性注射和骨质流失 （骨溶解）是由不良刺激诱导的，包括关节置换（JR）的磨损副产品。核 因子Kappa B（NF-κB）是巨噬细胞（M ϕ）和调节MSC的关键转录因子 炎症，骨形成/重塑和衰老。我们表明，将MSC通过脂肪垄断 - 糖（LPS）和肿瘤坏死因子-α（TNF-α）诱导NF-κB的急性瞬态激活 协同增强成骨的作用，并调节从促炎（M1）到一个的M ϕ极化 抗炎促再生（M2）表型。此外，使用使用持续的NF-κB信号抑制 NF-κB诱饵寡脱氧核苷酸（ODN）已显示可减轻炎症性骨质流失。我们有 转基因MSC以感知NF-κB激活，然后增加抗炎的产生 促增长细胞因子IL-4。这笔赠款的目的是加速净骨形成并减轻 通过NF-KB驱动免疫调节的慢性炎症和骨溶解，使用预处理或一般 修改后的MSC移植到本地环境。 特定目标＃1a-定义预处理与NF -κB感应的关键免疫调节相互作用 和IL-4分泌MSC在M ϕ上，暴露于骨科的磨损副产品。 特定目标＃1b-通过使用预处理与NF-κB传感和IL-4分泌来增强骨生成 MSC/M ϕ共培养模型中的MSC，暴露于骨科的磨损副产品。 特定目标＃2a-以证明移植预处理MSC对骨骼的治疗作用 在急性炎症阶段（模拟假体的初始实施阶段和 使用鼠连续股骨颗粒输注模型）。 具体目标＃2b-以证明预处理与NF-κB传感和IL-4的治疗作用 在慢性炎症阶段分泌净骨形成的MSC（模拟已建立的磨损 使用鼠连续股骨颗粒输注模型）。 特定目的＃3-证明调节急性和慢性炎症的上述原理是 在体内，有效的动物性别（雄性与雌性小鼠）或年龄（年轻小鼠）的有效性。我们假设 NF-κB预处理或NF-κB感应和IL-4分泌MSC将增强成骨和 MSC的免疫调节信号传导，通过Mϕ和MSC通过串扰增强骨形成。 NF-KB驱动器 MSC的免疫调节应减轻颗粒诱导的注射，破骨细胞激活并增强 年轻人和年龄，雄性和雌性小鼠的骨形成。 MSC的NF-KB驱动免疫调制是新颖的， 机械性，并直接转化为其他器官系统中的炎症性疾病。

项目成果

Preconditioning of murine mesenchymal stem cells synergistically enhanced immunomodulation and osteogenesis.

• DOI: 10.1186/s13287-017-0730-z
• 发表时间: 2017-12-06
• 期刊: Stem cell research & therapy
• 影响因子: 7.5
• 作者: Lin T;Pajarinen J;Nabeshima A;Lu L;Nathan K;Jämsen E;Yao Z;Goodman SB
• 通讯作者: Goodman SB

Role of direct estrogen receptor signaling in wear particle-induced osteolysis.

• DOI: 10.1016/j.biomaterials.2012.10.030
• 发表时间: 2013-01
• 期刊: BIOMATERIALS
• 影响因子: 14
• 作者: Nich, Christophe;Rao, Allison J.;Valladares, Roberto D.;Li, Chenguang;Christman, Jane E.;Antonios, Joseph K.;Yao, Zhenyu;Zwingenberger, Stefan;Petite, Herve;Hamadouche, Moussa;Goodman, Stuart B.
• 通讯作者: Goodman, Stuart B.

Contributions of human tissue analysis to understanding the mechanisms of loosening and osteolysis in total hip replacement.

• DOI: 10.1016/j.actbio.2014.02.003
• 发表时间: 2014-06
• 期刊: Acta biomaterialia
• 影响因子: 9.7
• 作者: Gallo J;Vaculova J;Goodman SB;Konttinen YT;Thyssen JP
• 通讯作者: Thyssen JP

Innate immunity sensors participating in pathophysiology of joint diseases: a brief overview.

• DOI: 10.1615/jlongtermeffmedimplants.2014010825
• 发表时间: 2014
• 期刊: Journal of long-term effects of medical implants
• 作者: Gallo J;Raska M;Konttinen YT;Nich C;Goodman SB
• 通讯作者: Goodman SB

Toll-like receptors-2 and 4 are overexpressed in an experimental model of particle-induced osteolysis.

• DOI: 10.1002/jbm.a.34972
• 发表时间: 2014-09
• 期刊: JOURNAL OF BIOMEDICAL MATERIALS RESEARCH PART A
• 影响因子: 4.9
• 作者: Valladares, Roberto D.;Nich, Christophe;Zwingenberger, Stefan;Li, Chenguang;Swank, Katherine R.;Gibon, Emmanuel;Rao, Allison J.;Yao, Zhenyu;Goodman, Stuart B.
• 通讯作者: Goodman, Stuart B.