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: 7989286
• 负责人: Elizabeth G Sweeney
• 金额: $ 11.89万
• 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 2013-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7989286
• 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; Cercopithecine Herpesvirus 1; 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; Medical; Medicine; Mentored Research Scientist Development Award; Mentors; Mitochondria; Modeling; Molecular; 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 Organisms; Transplantation; United States National Institutes of Health; Universities; Veterinary Medicine; Veterinary Schools; Wild Type Mouse; Work; angiogenesis; base; bone; cartilage matrix protein; cell type; congenic; cytokine; disease phenotype; in vivo; insight; interest; leukemia/lymphoma; long bone; member; mouse model; neonate; novel; perlecan; professor; progenitor; prophylactic; public health relevance; scaffold; skeletal; skeletogenesis; stem cell biology; stem cell niche; subcutaneous; substantia spongiosa; suburb; symposium; syndecan; vaccinia virus vector

DESCRIPTION (provided by applicant): 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. PUBLIC HEALTH RELEVANCE: This proposal will assess the contribution of skeletal development, involving the transition from cartilage to bone and marrow, to the establishment of the prerequisite marrow environment for hematopoiesis. We aim to identify extracellular matrix molecule(s) needed for blood cell differentiation using mouse models with skeletal and lymphopoietic defects. Data should yield insights into basic stem cell biology, as well as aid in diagnosis and treatment of skeleto-hematopoietic diseases.

描述（由申请人提供）：我在德雷克塞尔大学本科学习期间对细胞生物学和免疫学的复杂性产生了兴趣，在本科学习结束后，我在宾夕法尼亚大学传染病系工作，继续培养这种兴趣医学工程中的疾病并表征了用作活疫苗预防剂的各种新型牛痘病毒载体。然后，我继续在托马斯杰斐逊大学读研究生，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，研究乙型肝炎病毒的体内和体外免疫反应。在我的博士后研究期间，我已经能够进一步表征 X 胶原蛋白转基因小鼠和无效小鼠的淋巴细胞生成缺陷，并描述体外和体内细胞水平上寄生虫挑战的免疫反应缺陷。我的博士后工作成果是一个拟议的模型，描绘了软骨-骨连接处的造血生态位，我将在本次 K01 奖中进一步测试该模型。 该提案将是我博士后工作的延续，该工作描述了软骨内骨化（EO）（肥厚软骨被骨骼和骨髓取代）与造血生态位的建立之间的联系。为此，将使用胶原蛋白 X 转基因小鼠、基底膜蛋白亚型小鼠和 Syndecan-1 小鼠，因为我们提出，与胶原蛋白 X 小鼠一样，所有这些小鼠的疾病表型可能涉及生长板缺陷、骨髓环境改变和异常造血。我们提出 X 型胶原蛋白和硫酸乙酰肝素蛋白多糖，例如perlecan 和 syndecan-1 提供了一个结构网络，可以隔离肥大软骨中的细胞因子，网络破坏可能会导致软骨骨连接处 (COJ) 发生不适当的信号传导，从而导致造血缺陷。为了测试这一点，1) 骨髓移植、共培养和细胞因子分析将确认骨髓环境的改变，并识别无法支持胶原蛋白 X 小鼠造血的 COJ 驻留细胞类型（造口细胞、肥大软骨细胞或成骨细胞），2)异位骨形成测定将证实 EO 产生造血生态位，并且 X 型胶原蛋白小鼠在此过程中存在缺陷，并且 3) 流式细胞术和共培养测定将证实并开始表征基底膜蛋白亚形性和 Syndecan-1 无效小鼠的造血缺陷。这些数据是授予该研究者的 NRSA 已完成目标的延伸，并将为未来的目标提供数据，即表征细胞外基质对发育和疾病中富含细胞因子的生态位的贡献。 这些研究将在宾夕法尼亚大学进行，该大学是美国竞争性研究资助最多的接受者之一，每年获得超过 5 亿美元的外部资金。我将工作的具体研究环境是在兽医学院动物生物学系。成员是一个专门的基础科学小组，由约 20 名常务（助理至正任）教授和 7 名研究教授组成，所有这些教授均由 NIH 或 NSF 资助，并在生物化学、细胞、分子和发育等领域进行积极的研究生物学。研究对象包括骨骼发生、造血、转基因和精原细胞移植、克隆、线粒体、模式形成、染色质结构、血管生成、翻译后修饰的精氨酰化、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、压力、免疫学、眼睛发育和恐龙古生物学。教师和博士后研究员在院系研讨会上互动，进行良好的思想交流、技术援助和设施共享。由于我们位于宾夕法尼亚大学校园内，因此在兽医学院之外的互动机会很容易，其中包括医学院和牙科学院、威斯塔研究所以及费城儿童医院。此外，在宾夕法尼亚大学，我们有机会参加托马斯·杰斐逊大学、福克斯蔡斯大学、天普大学以及费城周边郊区的许多小型大学举办的校际研讨会和座谈会。最后，宾夕法尼亚大学的教职人员将为我提供一个独特的机会来寻求骨骼/基质生物学、血液学和 B 细胞生物学方面的建议，包括： Drs. Jacenko 和 Emerson（我的导师），博士。汉肯森、索斯洛夫斯基和亚当斯（在我的指导委员会中），博士。 Cancro 和 Akintoe（我的合作者），以及我将在宾夕法尼亚大学及周边学校提供的许多期刊俱乐部、研讨会和座谈会上与我互动的众多教职人员。 公共健康相关性：该提案将评估骨骼发育的贡献，包括从软骨到骨骼和骨髓的转变，以及建立造血的先决条件骨髓环境。我们的目标是利用具有骨骼和淋巴细胞生成缺陷的小鼠模型来鉴定血细胞分化所需的细胞外基质分子。数据应该可以深入了解基本的干细胞生物学，并有助于骨骼造血疾病的诊断和治疗。