Collagen X and heparan sulfate proteoglycan in the hematopoietic stem cell niche.

造血干细胞生态位中的 X 胶原蛋白和硫酸乙酰肝素蛋白多糖。

• 批准号: 8326237
• 负责人: Elizabeth G Sweeney
• 金额: $ 12.16万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-09-20 至 2012-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8326237
• 关键词: Affect; Americas; Animals; Aplastic Anemia; Attenuated Vaccines; Award; B-Lymphocytes; Basic Science; Binding; Biochemistry; Biological Assay; Biology; Blood Cells; Bone Marrow; Bone Marrow Cells; Bone Marrow Transplantation; Cartilage; Cell Cycle; Cell Death; Cell Differentiation process; Cells; Cellular biology; Chondrocytes; Chromatin Structure; Cloning; Coculture Techniques; Collagen; Collagen Type X; Communicable Diseases; Conditioned Culture Media; Data; Defect; Dental Schools; Development; Developmental Biology; Diagnosis; Differentiation and Growth; Dinosaurs; Disease; ENG gene; Elements; Engineering; Environment; Epiphysial cartilage; Exclusion; Exhibits; Extracellular Matrix; Eye Development; Faculty; Failure; Fellowship; Flow Cytometry; Fostering; Foxes; Funding; Future; Gene Silencing; Gene Transfer Techniques; Generations; Glycosaminoglycans; Goals; Growth Factor; Hematology; Hematopoiesis; Hematopoietic; Hematopoietic stem cells; Heparan Sulfate Proteoglycan; Hypocellular Bone Marrow; Immune System Diseases; Immune response; Immunity; Immunohistochemistry; Immunology; In Vitro; Institutes; Intercellular Junctions; Interleukin-3; Investigation; Journals; Knockout Mice; Link; Lymphopenia; Lymphopoiesis; Maintenance; Malignant Neoplasms; Marrow; Medicine; Mentored Research Scientist Development Award; Mentors; Mitochondria; Modeling; Molecular Biology; Mus; Mutation; National Institute of Arthritis and Musculoskeletal and Skin Diseases; National Research Service Awards; Obesity; Organ; Osteoblasts; Osteogenesis; Outcome; PTPRC gene; Pancytopenia; Parasites; Pattern Formation; Pediatric Hospitals; Pennsylvania; Perinatal; Phenotype; Philadelphia; Population; Post-Translational Protein Processing; Postdoctoral Fellow; Process; Protocols documentation; Research; Research Personnel; Research Project Grants; Respiration; Reverse Transcriptase Polymerase Chain Reaction; Rheumatoid Arthritis; Schools; Signal Transduction; Skeletal Development; Somatotropin; Spermatogenesis; Stem cells; Stress; Stromal Cells; Testing; Time; Transcriptional Regulation; Transgenes; Transgenic Mice; Transgenic Organisms; Transplantation; United States National Institutes of Health; Universities; Veterinary Medicine; Veterinary Schools; Virus; Wild Type Mouse; Work; angiogenesis; base; bone; cartilage matrix protein; cell type; congenic; cytokine; developmental immunology; disease phenotype; in vivo; insight; interest; leukemia/lymphoma; long bone; medical schools; member; mouse model; neonate; novel; perlecan; professor; progenitor; prophylactic; scaffold; skeletal; skeletogenesis; stem cell biology; stem cell niche; subcutaneous; substantia spongiosa; suburb; symposium; syndecan; vaccinia virus vector

I developed an interest in the complexities of cell biology and immunology during my undergraduate studies at Drexel University, which I continued to foster after my undergraduate studies, where I worked at the University of Pennsylvania in the Department of Infectious Diseases in Medicine engineering and characterized various novel Vaccinia virus vectors to be used as live vaccine prophylactics. I then went on to graduate school at Thomas Jefferson University where I broadened my studies to include general cell biology, developmental biology and immunology studying in vivo and in vitro immune responses to Hepatits B virus. During my post doctoral studies, I have been able to further characterize the lymphopoietic defects in the collagen X transgenic and null mice, as well as describe the immune response defects at both a cellular level in vitro and in vivo with parasite challenges. The outcome of my postdoctoral work is a proposed model depicting the hematopoietic niche in the cartilage-bone junction, which I will test further in this K01 award. This proposal will be a continuation of my post doctoral work characterizing the link between endochondral ossification (EO), where hypertrophic cartilage is replaced by bone and marrow, and the establishment of a hematopoietic niche. To do this, collagen X transgenic and null, perlecan hypomorphic and syndecan-1 null mice will be used, since we propose that, as in collagen X mice, the disease phenotypes of all these mice may involve growth plate defects, an altered marrow environment and aberrant hematopoiesis. We proposed that collagen X and heparan sulfate proteoglycans, e.g. perlecan and syndecan-1, provides a structural network that sequesters cytokines in hypertrophic cartilage, and network disruption may cause inappropriate signaling at the chondro-osseous junction (COJ) leading to defective hematopoiesis. To test this, 1) bone marrow transplants, co-cultures and cytokine analyses will confirm the altered marrow environment and identify the COJ resident cell type (stomal, hypertrophic chondrocyte, or osteoblast) unable to support hematopoiesis in the collagen X mouse, 2) ectopic bone forming assays will confirm EO generates the hematopoietic niche and that collagen X mice are defective in this process, and 3) flow cytometry and co-culture assays will confirm and begin to characterize the hematopoietic defects in the perlecan hypomorphic and syndecan-1 null mice. These data are an extension to the completed aims of a NRSA awarded to this investigator, and will provide data towards future goals of characterizing the extracellular matrix contribution to cytokine rich niches in development and disease. These studies will be performed at the University of Pennsylvania, which is ranked among America's top recipients of competitive research grants, earning more than $500 million in external funding annually. The specific research environment that I will be working is in the Department of Animal Biology at the School of Veterinary Medicine. The members are an exclusively basic science group, consisting of ~20 standing (Assistant to Full) professors, and 7 research professors, all of who are funded by NIH or NSF, and are conducting active research in biochemistry, cell, molecular, and developmental biology. Subjects under investigation include skeletogenesis, hematopoiesis, transgenesis and spermatogonial transplanation, cloning, mitochondria, pattern formation, chromatin structure, angiogenesis, arginylation as a posttranslational modification, transcriptional regulation of differentiation, growth hormones, respiration, cell cycle, cell death, spermatogenesis, obesity, stress, immunology, eye development, and dinosaur paleobiology. The faculty and postdoctoral fellows interact at department seminars, and there is good exchange of ideas, technical assistance, and sharing of facilities. Opportunities for interactions outside of the Veterinary School are effortless, since we are within the University of Pennsylvania campus, which includes the Medical and Dental Schools, Wistar Institute, as well as Children's Hospital of Philadelphia. Further, at the University of Pennsylvania we have the opportunity to participate in may intercollegiate seminars and symposiums with Thomas Jefferson University, Fox Chase, Temple University, and the many small collages in the surrounding suburbs of Philadelphia. Finally, the Penn faculty members that will provide me with a unique opportunity to seek advise in skeletal/matrix biology, hematology, and B cell biology, including: Drs. Jacenko and Emerson (my mentors), Drs. Hankenson, Soslowsky, and Adams (on my mentoring committee), Drs. Cancro and Akintoe (my collaborators), and the numerous faculty members that I will interact with at the many journal club, seminar, and symposiums offered at Penn and the surrounding schools.

在德雷塞尔大学(Drexel University)读本科期间，我对细胞生物学和免疫学的复杂性产生了兴趣，本科学业结束后，我继续培养这种兴趣，在那里我在宾夕法尼亚大学医学工程传染病系工作，并表征了各种可用作活疫苗预防的新型痘苗病毒载体。然后我去了托马斯·杰斐逊大学研究生院，在那里我扩大了我的研究范围，包括普通细胞生物学、发育生物学和免疫学，研究体内和体外对乙肝病毒的免疫反应。在我的博士后研究期间，我能够进一步描述X胶原蛋白转基因小鼠和空白小鼠的淋巴生成缺陷，以及在体外和体内对寄生虫挑战的细胞水平的免疫反应缺陷。我博士后工作的成果是一个拟议的模型，描述了软骨-骨交界处的造血生态位，我将在这个K01奖中进一步测试。 这项建议将是我博士后工作的继续，该工作描述了软骨内骨化(EO)和建立造血利基之间的联系。软骨内骨化是指肥大的软骨被骨骼和骨髓取代。要做到这一点，将使用X型胶原转基因小鼠和缺失、Perlecan亚型和syndecan-1缺失的小鼠，因为我们认为，就像X型胶原小鼠一样，所有这些小鼠的疾病表型都可能涉及生长板缺陷、骨髓环境改变和异常造血。我们认为，X型胶原和硫酸肝素蛋白多糖，如Perlecan和Syndecan-1，提供了一个结构网络，将细胞因子隔离在肥大的软骨中，而网络的破坏可能会导致软骨-骨连接(Coj)处不适当的信号传递，导致造血功能障碍。为了证明这一点，1)骨髓移植、共培养和细胞因子分析将确认X胶原蛋白缺失小鼠的骨髓环境发生了变化，并鉴定了COJ驻留细胞类型(造口、肥大的软骨细胞或成骨细胞)无法支持X胶原蛋白的造血；2)异位成骨检测将证实EO产生了造血龛并且X胶原蛋白小鼠在此过程中存在缺陷；3)流式细胞仪和共培养分析将证实并开始鉴定Perlecan亚型和syndecan-1基因缺失小鼠的造血缺陷。这些数据是授予这位研究人员的NRSA已完成目标的延伸，并将为未来确定细胞外基质对发育和疾病中丰富的细胞因子利基的贡献的目标提供数据。 这些研究将在宾夕法尼亚大学进行，该大学是美国最具竞争力的研究拨款获得者之一，每年获得超过5亿美元的外部资金。我将要工作的具体研究环境是在兽医学院的动物生物学系。成员是一个专门的基础科学小组，由大约20名常务(助理到全职)教授和7名研究教授组成，他们都是由NIH或NSF资助的，从事生物化学、细胞、分子和发育生物学的活跃研究。正在研究的主题包括骨骼发生、造血、移植和精原移植、克隆、线粒体、图案形成、染色质结构、血管生成、作为翻译后修饰的精氨酸化、分化的转录调控、生长激素、呼吸、细胞周期、细胞死亡、精子发生、肥胖、应激、免疫学、眼睛发育和恐龙古生物学。教员和博士后研究员在系研讨会上进行互动，并进行良好的思想交流、技术援助和设施共享。在兽医学院之外进行互动的机会是毫不费力的，因为我们在宾夕法尼亚大学校园内，其中包括医学和牙科学校、Wistar研究所以及费城儿童医院。此外，在宾夕法尼亚大学，我们有机会参加5月与托马斯·杰斐逊大学、福克斯·蔡斯大学、坦普尔大学以及费城郊区许多小型大学举行的校际研讨会和研讨会。最后，我要感谢宾夕法尼亚大学教职员工，他们将为我提供在骨骼/基质生物学、血液学和B细胞生物学方面的独特建议，包括：Jacenko和Emerson博士(我的导师)、Hankenson博士、Soslowsky博士和亚当斯博士(我的导师委员会成员)、Cancro博士和Akintoe博士(我的合作者)，以及我将在宾夕法尼亚大学和周边学校提供的许多杂志俱乐部、研讨会和座谈会上与之互动的众多教职员工。