The Hematopoietic Stem Cell Expansion: Role of Early T Lymphocytes

造血干细胞扩增：早期 T 淋巴细胞的作用

• 批准号: 8081801
• 负责人: Aleksandar Milan Babic
• 金额: $ 13.47万
• 依托单位: METHODIST HOSPITAL RESEARCH INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-07-14 至 2012-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8081801
• 关键词: Adult; Allogenic; Anemia; Angiopoietins; Apoptosis; Autoimmune Diseases; Biological Assay; Blood Cells; Bone Marrow; Bone Marrow Transplantation; CD3 Antigens; CD4 Positive T Lymphocytes; Cell Count; Cell Death; Cell Surface Proteins; Cell model; Cells; Coculture Techniques; Confocal Microscopy; Cultured Cells; Data; Development; Disease; Embryo; Engraftment; Fetal Liver; Fibroblast Growth Factor 1; Frozen Sections; Gene Expression; Gene Expression Profile; Genes; Glycolipids; Goals; Grant; Growth Factor; Hematologic Neoplasms; Hematopoietic stem cells; Hepatocyte; Insulin-Like Growth Factor II; Laboratories; Lead; Liver; Lymphocyte; Malignant lymphoid neoplasm; Marrow; Measures; Memory; Microarray Analysis; Molecular Models; Mus; Patients; Pattern; Phenotype; Population; Procedures; Proteins; Research Personnel; Role; SLAM protein; Serum; Serum-Free Culture Media; Signal Transduction; Sorting - Cell Movement; Spleen; Staining method; Stains; Stem Cell Factor; Stromal Cells; Supporting Cell; Surface; System; T-Cell Receptor; T-Lymphocyte; Testing; Thrombopoietin; Time; Tissues; Umbilical Cord Blood Transplantation; Work; abstracting; bone; cell type; clinical practice; fetal; genetic regulatory protein; improved; in vivo; killer T cell; molecular modeling; notch protein; novel; novel therapeutic intervention; reconstitution; research study; self-renewal; spatial relationship; trend

DESCRIPTION (provided by applicant): Bone marrow transplantation (BMT) is used to treat a variety of diseases including hematological and lymphoid malignancies, anemias, autoimmune disorders and others. The critical component of each BMT is hematopoietic stem cells (HSC), which are defined by their ability to self-renew and to differentiate into all blood cell types. The objective of this grant is to understand the role of early T cells in regulating HSC selfrenewal and expansion. Recent studies from the Lodish laboratory showed that coculture of embryonic day 15.5 (E15.5) mouse fetal liver (FL) CD3+ cells with enriched adult bone marrow HSCs led to expansion of HSC numbers and activities. These CD3+ cells specifically express several secreted growth factors including Insulin-like Growth factor 2 (IGF-2) and Angiopoietin-like 2 and 3 (Angptl 2 and 3). Inclusion of these factors in a serum-free culture medium also containing Stem Cell Factor (SCF), Thrombopoietin (TPO), and Fibroblast Growth Factor 1 supported a 24 to 30 fold net expansion of long-term HSC, as observed by reconstitution analysis. In preliminary studies we showed that significant proportion of E15.5 FL CD3+ cells are CD4" and CDS" but positive for either T cell receptor ap or y6 and thus likely are early T cells. In this proposal I will: 1) Characterize phenotype and activation/memory status of subpopulations of CD3+ cells in E15.5 FL, and test the hypothesis that these cells are early natural killer T cells; examine ability of subpopulations of FL cells to support ex vivo self-renewal and expansion of HSCs; examine expression pattern of genes known to promote self-renewal of HSCs in the subpopulations of CD3+ cells that promote ex vivo self-renewal and expansion of HSCs. 2) Examine adult bone marrow, spleen and liver for the presence of subpopulations of CD3+ cells similar to subpopulations of E15.5 FL CD3+ cells that are capable of ex vivo expansion of BM HSCs. 3) Examine gene expression profile of purified FL and adult CD3+ cell populations that support HSC expansion in culture using microarray analysis, to identify potentially novel proteins with HSC supportive function. The clinical practice data suggests that the number of patients that are potential candidates for BMT has increased significantly in the past decade while the availability of compatible donors did not follow this trend. Our capability to expand the number of available HSC, would not only lead to increase in the number of potential donors and improved engraftment, but it would also promote development of new therapeutic approaches using HSC manipulation. (End of Abstract)

描述（由申请人提供）： 骨髓移植（BMT）用于治疗各种疾病，包括血液学和 淋巴恶性肿瘤，贫血，自身免疫性疾病等。每个BMT的关键成分是造血干细胞（HSC），它们的定义是它们的自我更新和分化为所有血细胞类型的能力。这项赠款的目的是了解早期T细胞在调节HSC自我繁殖和扩张中的作用。来自Lodish实验室的最新研究表明，胚胎第15.5（E15.5）小鼠胎儿肝脏（FL）CD3+细胞具有富集的成年骨髓HSC，导致HSC数量和活动的扩大。这些CD3+细胞特别表达了几种分泌的生长因子，包括胰岛素样生长因子2（IGF-2）和类似血管素的2和3（ANGPTL 2和3）。如重组分析所观察到的那样，将这些因子纳入无血清培养基中还包含干细胞因子（SCF），血小板蛋白（TPO）和成纤维细胞生长因子1支持长期HSC的24至30倍净膨胀，如重新建立分析所观察到的。在初步研究中，我们表明，E15.5 FL CD3+细胞的显着比例为CD4和CDS，但对于T细胞受体AP或Y6呈阳性，因此可能是早期T细胞。在此提案中，我将：1）表征E15.5 FL中CD3+细胞亚群的表型和激活/记忆状态，并检验这些细胞是早期天然杀伤剂T细胞的假设；检查FL细胞亚群支持离体自我更新和HSC扩展的能力；检查已知的基因的表达模式，这些基因已知促进HSC在CD3+细胞的亚群中的自我更新，从而促进了体内自我更新和HSC的扩张。 2）检查成年骨髓，脾脏和肝脏是否存在类似于能够在BM HSC的E15.5 FL CD3+细胞亚群的CD3+细胞的亚群。 3）检查使用微阵列分析支持HSC扩展的纯化FL和成年CD3+细胞群的基因表达谱，以鉴定具有HSC支持功能的潜在新型蛋白质。临床实践数据表明，在过去的十年中，BMT潜在候选者的患者数量显着增加，而兼容供体的可用性并未遵循这一趋势。我们扩大可用HSC数量的能力，不仅会导致潜在的捐助者数量和改善的植入量增加，而且还将通过HSC操纵来促进新的治疗方法的开发。 （抽象的结尾）

项目成果

Culture of mouse amniotic fluid-derived cells on irradiated STO feeders results in the generation of primitive endoderm cell lines capable of self-renewal in vitro.

在经过辐射的 STO 饲养层上培养小鼠羊水来源的细胞，可产生能够在体外自我更新的原始内胚层细胞系。

• DOI: 10.1159/000353942
• 发表时间: 2013
• 期刊: Cells, tissues, organs
• 作者: Babic,AleksandarM;Jang,Sunyoung;Nicolov,Eugenia;Voicu,Horatiu;Luckey,ChanceJ
• 通讯作者: Luckey,ChanceJ