BLR&D Research Career Scientist Award

BLR

• 批准号: 10265365
• 负责人: Kin-Hing William Lau
• 金额: --
• 依托单位: VA LOMA LINDA HEALTHCARE SYSTEM
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-04-01 至 2022-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10265365
• 关键词: Age; Aging; Area; Award; Binding; Biogenesis; Bone Development; Bone Diseases; Bone Resorption; Calcium; Cartilage; Cells; ChIP-seq; Chimeric Proteins; Chondrocytes; Chondrogenesis; Complex; DNA cassette; Degenerative polyarthritis; Development; Disease Progression; Enzymes; Epigenetic Process; Etiology; Female; Foundations; Fracture; Functional disorder; Funding; Genes; Genetic Transcription; Grant; Grant Review; Health; Health Care Costs; Hip Fractures; Human; In Vitro; Incidence; Inflammatory; Intra-Articular Injections; Investigation; Joints; Knee; Knockout Mice; Knowledge; Lead; Ligands; Mediating; Medical Care Costs; Messenger RNA; MicroRNAs; Military Personnel; Mission; Molecular; Morbidity - disease rate; Mus; Natural regeneration; Osteoclasts; Osteogenesis; Osteopenia; Osteoporosis; Osteoporosis prevention; Ovariectomy; Pain; Pathologic; Pathology; Pathway interactions; Patient Care; Patients; Phenotype; Phosphotyrosine; Physiological; Play; Population; Protein Overexpression; Protein Tyrosine Kinase; Protein Tyrosine Phosphatase; Proteins; Quality of life; Regimen; Regulation; Research; Rho-associated kinase; Role; Scientist; Signal Transduction; Synovial Membrane; Testing; Therapeutic; Transgenes; Transgenic Mice; United States National Institutes of Health; Up-Regulation; Veterans; Wasting Syndrome; Work; active duty; age related; arthropathies; articular cartilage; base; bone; bone loss; bone mass; career; chondrogenesis factor; conditional knockout; cytokine; disabling symptom; effective therapy; improved; in vivo; innovation; insight; interest; joint injury; macrophage; monocyte; new therapeutic target; novel; novel strategies; novel therapeutics; osteoporotic bone; overexpression; patient population; prevent; promoter; protein expression; screening; skeletal tissue; src-Family Kinases; subchondral bone; therapeutically effective; transcription factor

The current research focus of the nominee is two-fold: The first is to delineate a unique molecular mechanism regulating functional activity of osteoclasts that involves negative regulation of the expression of a distinctive osteoclastic protein-tyrosine phosphatase (PTP-oc), which is a potent activator of functional activity of mature osteoclasts, by microRNA-17 (miR17). This project is supported by a BLR&D Merit Review award and seeks to test 3 hypotheses: 1) resorption cytokines activates osteoclasts via suppression of miR17 expression; 2) conditional deletion of miR17 in osteoclasts increases PTP-oc expression, which in turn stimulates osteoclast activity via up-regulation of the PTP-oc signaling; and 3) conditional overexpression of miR17 in osteoclasts reduces PTP-oc expression and inhibits physiologically- and pathologically-induced bone resorption. It has three Aims: Aim 1 tests the first hypothesis by determining if treatment with resorption modulators regulates the promoter activity of miR17 in osteoclasts; demonstrating direct effects of resorption modulators on transcription of the miR17~92 gene; and performing ChIP-seq analysis to identify key transcription factors. Aim 2 tests the second hypothesis by characterizing the in vivo and in vitro bone and osteoclast phenotypes of osteoclast miR17~92 conditional knockout mice, determining effects of miR17~92 deficiency on PTP-oc mRNA level, the PTP-oc signaling, and resorption activity of osteoclasts, and determining whether re-introduction of miR17 into miR17~92 deficient osteoclasts would “restore” the osteoclast phenotype; and evaluating effects of miR17~92 deficiency on osteoclast differentiation. Aim 3 tests the third hypothesis by generating transgenic mice with conditional overexpression of miR17~92 in osteoclasts by crossing LysM-Cre mice with mice harboring the miR17~92 transgene downstream to a loxP-flanked Neo-STOP cassette at the ROSA locus, determining the effects of conditional overexpression of miR17~92 in osteoclasts on bone and osteoclast phenotypes, and if overexpression of miR17~92 in osteoclasts would blunt the ovariectomy- and calcium depletion-induced bone resorption. This work should yield insights into how mature osteoclast activity is regulated, which is important in our overall understanding of the pathophysiology of various subtypes of osteoporosis and bone-wasting diseases. It may also provide novel drug targets (e.g., miR17) for screening of small molecular compounds for use to develop effective anti-resorption therapies for certain subtypes of osteoporosis. The second focus is to develop an innovative EphA4-based small molecular therapeutic strategy to prevent and treat osteoarthritis (OA) and posttraumatic OA (PTOA). This project is funded by a DoD Idea Development award. The rationale of this project is based on the exciting discovery of an opposing regulatory effects of the forward signaling of EphA4 on osteoclasts (inhibiting bone resorption) as opposed to on chondrocytes (promoting cartilage biogenesis). This project seeks to capitalize these unique opposing actions of the EphA4 forward signaling on osteoclasts vs. chondrocytes to develop a novel strategy that would simultaneously reduce degradation of articular cartilage/subchondral bone and enhance regeneration of articular cartilage. It has three Aims. Aim 1 determines whether treatment of monocytes/macrophages/synoviocytes isolated from PTOA synovium with soluble EfnA4-fc protein suppresses secretion of pro-inflammatory cytokines and degradative enzymes, and if treatment of articular chondrocytes with EfnA4-fc would promote chondrocyte proliferation, maturation, and survival in vitro. Aim 2 seeks to develop and optimize an EphA4-binding EfnA-fc-based strategy, which involves direct intra-articular injection of a soluble EfhA-fc chimeric protein into the OA/PTOA joint. Aim 3 determines if administration of the optimized EfnA-fc-based strategy to the PTOA/OA synovium joint shortly after the creation of an intra-articular tibial plateau fracture in the knee prevents degradation of articular cartilage, and if this optimized EfnA-fc-based therapeutic strategy promotes regeneration of articular cartilage. This work should provide feasibility evidence for this EphA4-based therapeutic therapy for PTOA/OA.

被提名人目前的研究重点有两个：第一是勾画出独特的分子机制 调节破骨细胞的功能活性，其涉及特异性的 破骨细胞蛋白-酪氨酸磷酸酶（PTP-oc）是成熟细胞功能活性的有效激活剂 microRNA-17（miR 17）检测破骨细胞。该项目由BLR&D Merit Review奖支持，旨在 检验3种假设：1）再吸收细胞因子通过抑制miR 17表达激活破骨细胞; 2） 破骨细胞中miR 17的条件性缺失增加PTP-oc表达，这反过来刺激破骨细胞 通过上调PTP-a信号传导的活性;和3）破骨细胞中miR 17的条件性过表达 降低PTP-α表达并抑制生理学和病理学诱导的骨吸收。它有 三个目的：目的1通过确定用再吸收调节剂治疗是否调节了 miR 17在破骨细胞中的启动子活性;证明了再吸收调节剂对转录的直接作用 miR 17 ~92基因的转录调控;以及进行ChIP-seq分析以鉴定关键转录因子。目标2测试 第二种假设，通过表征破骨细胞的体内和体外骨和破骨细胞表型 miR 17 ~92条件性基因敲除小鼠，测定miR 17 ~92缺陷对PTP-oc mRNA水平的影响， PTP-a信号传导和破骨细胞的再吸收活性，并确定是否将miR 17重新引入破骨细胞中。 miR 17 ~92缺陷破骨细胞将“恢复”破骨细胞表型; 破骨细胞分化不足。目的3通过产生转基因小鼠来测试第三种假设， 通过LysM-Cre小鼠与携带miR 17 ~92的小鼠杂交，在破骨细胞中条件性过表达miR 17 ~92。 将miR 17 ~92转基因下游的ROSA基因座处的loxP-侧翼的Neo-STOP盒，决定了 破骨细胞中miR 17 ~92的条件性过表达对骨和破骨细胞表型的影响，以及 miR 17 ~92在破骨细胞中的过表达会使卵巢切除和钙缺乏诱导的骨变钝 再吸收这项工作将有助于深入了解成熟破骨细胞活性是如何调节的，这在 我们对骨质疏松症和骨消耗的各种亚型的病理生理学的总体理解 疾病它还可以提供新的药物靶点（例如，miR 17）用于筛选小分子化合物， 用于开发针对某些骨质疏松亚型的有效抗吸收疗法。第二个重点是， 开发基于EphA 4的创新小分子治疗策略，以预防和治疗骨关节炎 (OA)创伤后OA（PTOA）。该项目由DoD创意开发奖资助。的理由 该项目的基础是令人兴奋的发现，即前向信号的相反调节作用， EphA 4在破骨细胞上（抑制骨吸收），而不是在软骨细胞上（促进软骨 生物发生）。该项目旨在利用EphA 4正向信号传导的这些独特的相反作用， 破骨细胞与软骨细胞，以开发一种新的策略，同时减少降解的 关节软骨/软骨下骨和增强关节软骨的再生。它有三个目标。要求1 确定是否用以下物质处理从PTOA滑膜分离的单核细胞/巨噬细胞/滑膜细胞 可溶性EfnA 4-fc蛋白抑制促炎细胞因子和降解酶的分泌，如果 用EfnA 4-fc处理关节软骨细胞将促进软骨细胞增殖、成熟和 体外存活目的2寻求开发和优化基于EphA 4结合EfnA-fc的策略，其 包括将可溶性EfhA-fc嵌合蛋白直接关节内注射到OA/PTOA关节中。目标3 确定在PTOA/OA滑膜关节短期内给予优化的基于EfnA-fc的策略是否 在膝关节内形成胫骨平台骨折后， 如果这种优化的基于EfnA-fc的治疗策略促进了关节软骨的再生， 这项工作将为基于EphA 4的PTOA/OA治疗提供可行性证据。

项目成果

In Vivo Generation of Gut-Homing Regulatory T Cells for the Suppression of Colitis.

• DOI: 10.4049/jimmunol.1800018
• 发表时间: 2019-06-15
• 期刊: Journal of immunology (Baltimore, Md. : 1950)
• 作者: Xu Y;Cheng Y;Baylink DJ;Wasnik S;Goel G;Huang M;Cao H;Qin X;Lau KW;Chan C;Koch A;Pham LH;Zhang J;Li CH;Wang X;Berumen EC;Smith J;Tang X
• 通讯作者: Tang X

Effect of aging on stem cells.

• DOI: 10.5493/wjem.v7.i1.1
• 发表时间: 2017-02-20
• 期刊: World journal of experimental medicine
• 作者: Ahmed AS;Sheng MH;Wasnik S;Baylink DJ;Lau KW
• 通讯作者: Lau KW

Dendritic cells, engineered to overexpress 25-hydroxyvitamin D 1α-hydroxylase and pulsed with a myelin antigen, provide myelin-specific suppression of ongoing experimental allergic encephalomyelitis.

• DOI: 10.1096/fj.201601243r
• 发表时间: 2017-07
• 期刊: FASEB journal : official publication of the Federation of American Societies for Experimental Biology
• 作者: Li CH;Zhang J;Baylink DJ;Wang X;Goparaju NB;Xu Y;Wasnik S;Cheng Y;Berumen EC;Qin X;Lau KW;Tang X
• 通讯作者: Tang X

1,25-Dihydroxyvitamin D suppresses M1 macrophages and promotes M2 differentiation at bone injury sites.

• DOI: 10.1172/jci.insight.98773
• 发表时间: 2018-09
• 期刊: JCI insight
• 影响因子: 8
• 作者: Samiksha Wasnik;C. Rundle;David J. Baylink;M. Yazdi;Edmundo E. Carreon;Yi Xu;X. Qin;K. Lau