BONE MARROW DERIVED FIBROBLASTS AND THEIR ROLE IN TISSUE REPAIR AND FIBROSIS

骨髓来源的成纤维细胞及其在组织修复和纤维化中的作用

• 批准号: 8167640
• 负责人: MEHRDAD ABEDI
• 金额: $ 33.49万
• 依托单位: ROGER WILLIAMS HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-05-01 至 2011-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8167640
• 关键词: Bleomycin; Bone Marrow; California; Cell Lineage; Cell surface; Cells; Centers of Research Excellence; Characteristics; Cicatrix; Computer Retrieval of Information on Scientific Projects Database; Data; Development; Differentiation Antigens; Dimensions; Doctor of Philosophy; Endomyocardial Fibrosis; Faculty; Fatal Outcome; Fibroblasts; Fibrosis; Flow Cytometry; Funding; Genes; Grant; Hematopoietic; Human; Immunologist; Injury; Institution; LacZ Genes; Left; Liver Fibrosis; Marrow; Methods; Modeling; Monitor; Mus; Myelofibrosis; Natural regeneration; Organ; Population; Process; Productivity; Pulmonary Fibrosis; Recruitment Activity; Research; Research Personnel; Resources; Role; Sclerosing Cholangitis; Site; Skin; Source; Stem cells; Subgroup; Systemic disease; T-Lymphocyte; Techniques; Thyroiditis; Time; Tissue Differentiation; Tissue Model; Tissues; Transplantation; United States National Institutes of Health; Universities; Work; Wound Healing; graft vs host disease; injured; lung melanoma; lymph nodes; tumor

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. There has been no change in the scope of this project. The project remains focused on tissue repair, with a particular emphasis on fibrosis. The project leader, Dr. Abedi, left the institution for a major promotion and to be on the faculty at the University of California at Davis. In the interim period, the PI of the COBRE, Dr. Vincent Falanga, took over the responsibility of maintaining this project at a high level of productivity in terms of progress and scienfic fundings. The NIH were properly notified of this change. A new PI (Ashleigh Boyd, PhD) has been recruited from Oxford University and has now taken over the project. Dr. Boyd is both an immunologist and a stem cell investigator, and thus brings new ideas to the project. For now, however, no change in hypothesis and specific aims have been made. Below is a narrative of the original project description. Excessive fibrosis is the prominent histological features of many human systemic diseases such as idiopathic and secondary pulmonary fibrosis, myelofibrosis, endomyocardial fibrosis, sclerosing cholangitis, hepatic fibrosis, cirrohsis and fibrous thyroiditis. Another important dimension is the fibrosis that occurs with tumors. Overall, the fibrosis in many conditions results in a severe, and in most cases, irreversible loss of organ function. The fatal outcome of many of these conditions can be directly attributed to the fibrotic process. Recent data has suggested that marrow derived cells can significantly contribute to the fibroblasts both in normal regeneration process and also in excessive fibrosis in pathological conditions. Many of the investigators believe that the stromal component of the marrow is responsible for these marrow derived fibroblasts. However our recent data shows that cells with hematopoietic characteristics are able to produce fibroblasts in the injured skin. Our hypothesis is that fibroblasts in the scar tissue and also in skin models of excessive fibrosis are originate from the hematopoietic component of bone marrow. The current study will be undertaken to investigate whether the increase in fibroblasts at the site of pathogenic fibrosis originated from transplanted hematopoietic donor cells or they are mostly coming from the residing fibroblasts in the adjacent tissue. We are also planning to establish a hierarchical model of differentiation for tissue fibroblast by identifying differentiation markers from marrow stem cells to fully differentiated fibroblasts. We have proposed the following specific aims for this project: 1) To characterize a specific subgroup of marrow cells that is able to contribute to tissue fibroblasts in the scar tissue. To induce fibrosis, we will use three well defined model of tissue fibrosis, ie. bleomycin injury, graft versus host disease, and TSK mice; 2) To characterize the role of marrow derived fibroblasts in comparison to their endogenous counterparts in each specific model of tissue fibrosis. We will determine whether marrow derived fibroblasts, both quantitatively and qualitatively, contribute to the fibrosis process. A main component of the proposed work is that FSP1-GFP, ColA2-LacZ mice will be used as functional models to identify the differentiation of marrow cells to fibroblasts. 3) To identify the developmental steps in the process of differentiation of bone marrow derived fibroblasts. Donor derived fibroblast cells residing in the recipient scar tissue will be isolated and analyzed by gene microarray and the results will be compared to the marrow cells from which they were derived and also de novo fibroblasts from the same scar. The genes identified by this method will be confirmed with a combination of real time-PCR, immunofluorescent and flow cytometry techniques, the progeny and lineages of these cells. Emphasis will be placed on establishing an immunophenotypic profile for marrow derived fibroblasts. The progressive loss/addition of cell surface markers during fibroblast maturation will be monitored, as it is done with the step-wise maturation of other populations of marrow cells, such as B or T lymphocytes. 4) The role of marrow derived fibroblasts in the fibrosis observed in tumors. During tenure of this grant we plan to look very early on tumors from lung, melanoma, and lymph nodes.

这个子项目是许多利用 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。列出的机构为 中心，但不一定是研究者所在的机构。 该项目的范围没有变化。该项目仍然专注于组织修复，特别强调纤维化。项目负责人阿贝迪博士离开了该机构，获得了一次重大晋升，成为加州大学戴维斯分校的教员。在此期间，COBRE的PI Vincent Falanga博士接管了在进度和科学基金方面保持该项目高水平生产力的责任。美国国立卫生研究院已适当地通知了这一变化。一个新的PI（Ashleigh Boyd，博士）已从牛津大学招募，并已接管该项目。博伊德博士既是一名免疫学家，也是一名干细胞研究者，因此为该项目带来了新的想法。然而，到目前为止，假设和具体目标都没有改变。 以下是原始项目说明的叙述。 过度纤维化是许多人类系统性疾病的突出组织学特征，如特发性和继发性肺纤维化、骨髓纤维化、肌内膜纤维化、硬化性胆管炎、肝纤维化、肝硬化和纤维性甲状腺炎。另一个重要的方面是与肿瘤一起发生的纤维化。总的来说，在许多情况下，纤维化导致严重的，并且在大多数情况下，不可逆的器官功能丧失。许多这些疾病的致命结果可以直接归因于纤维化过程。最近的数据表明，骨髓来源的细胞可以显着有助于成纤维细胞在正常的再生过程中，也在过度的纤维化的病理条件。许多研究者认为，骨髓的基质成分是这些骨髓来源的成纤维细胞的原因。然而，我们最近的数据表明，具有造血特性的细胞能够在受伤的皮肤中产生成纤维细胞。我们的假设是，瘢痕组织和过度纤维化的皮肤模型中的成纤维细胞起源于骨髓的造血成分。目前的研究将调查致病性纤维化部位的成纤维细胞的增加是否来源于移植的造血供体细胞，或者它们主要来自邻近组织中驻留的成纤维细胞。我们还计划通过鉴定从骨髓干细胞到完全分化的成纤维细胞的分化标志物来建立组织成纤维细胞分化的分级模型。 我们提出了以下具体目标：1）表征能够在瘢痕组织中形成组织成纤维细胞的特定骨髓细胞亚群。为了诱导纤维化，我们将使用三种明确定义的组织纤维化模型，即。博来霉素损伤、移植物抗宿主病和TSK小鼠; 2）在每种特定的组织纤维化模型中，与其内源性对应物相比，表征骨髓来源的成纤维细胞的作用。我们将确定是否骨髓来源的成纤维细胞，定量和定性，有助于纤维化过程。所提出的工作的一个主要组成部分是，FSP 1-GFP，ColA 2-LacZ小鼠将被用作功能模型，以确定骨髓细胞向成纤维细胞的分化。3)目的：明确骨髓成纤维细胞分化过程中的发育步骤。将存在于受体瘢痕组织中的供体来源的成纤维细胞分离并通过基因微阵列进行分析，并将结果与其来源的骨髓细胞以及来自相同瘢痕的新生成纤维细胞进行比较。通过该方法鉴定的基因将用真实的时间PCR、免疫荧光和流式细胞术技术、这些细胞的后代和谱系的组合来确认。重点将放在建立骨髓来源的成纤维细胞的免疫表型。将监测成纤维细胞成熟期间细胞表面标志物的进行性丢失/添加，如其他骨髓细胞群（如B或T淋巴细胞）的逐步成熟。4)骨髓源性成纤维细胞在肿瘤纤维化中的作用。在该基金的任期内，我们计划对来自肺部、黑色素瘤和淋巴结的肿瘤进行早期研究。