Role of Endothelial Progenitors in Hematopoietic Stem Cell Expansion

内皮祖细胞在造血干细胞扩增中的作用

• 批准号: 7525677
• 负责人: Mervin C. YODER
• 金额: $ 23.09万
• 依托单位: UNIVERSITY OF CALIFORNIA AT DAVIS
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/7525677
• 关键词: Adult; Age; Aging; Animals; Authorization documentation; Blood; Blood Vessels; Blood capillaries; Blood specimen; Budgets; CD34 gene; Categories; Cell Line; Cell Surface Proteins; Cell Transplantation; Cells; Clinical; Coculture Techniques; Commit; Data; Data Analyses; Development; Direct Costs; Disclosure; Endothelial Cells; Endothelium; Engraftment; Equipment; Face; Facilities and Administrative Costs; Fees; Frequencies; Goals; Growth; Growth Factor; Hematopoietic; Hematopoietic System; Hematopoietic stem cells; Human; Human Resources; In Vitro; Indiana; Injection of therapeutic agent; Inpatients; Instruction; Kidney; Laboratories; Last Name; Lead; Macaca mulatta; Marrow; Mesenchymal Stem Cells; Methods; Modeling; Morphology; Names; Numbers; Outpatients; PTPRC gene; Patient Care; Patients; Population; Pre-Clinical Model; Principal Investigator; Procedures; Property; Registries; Research; Research Personnel; Role; SCID Mice; Sampling; Secondary to; Source; Staging; Stem cells; Structure; Telomerase; Testing; Time; Transplantation; Travel; Umbilical Cord Blood; Umbilical cord structure; Universities; Vascular Endothelium; Wages; aged; base; capillary; cost; cytokine; day; feeding; fetal; human embryonic stem cell; improved; in utero; novel; peripheral blood; progenitor; programs

Human umbilical cord blood has been demonstrated to be an effective source of hematopoietic stem cells (HSC) for clinical transplantation. A major limitation to the more widespread use of this material for adult patients is that the number of HSC in a cord blood sample may be insufficient to engraft in adult patients, particularly unrelated and mismatched recipients. Determination of a method to expand HSC in cord blood would be a major advance in the field. We have recently identified a hierarchy of endothelial progenitor cells (EPC) that circulate in the blood, but also comprise a portion of the endothelial cell intima in human and rhesus macaque blood vessels. Similar to the hematopoietic system, EPC are organized in a hierarchy in which the most proliferative progenitors give rise to numerous progeny with less proliferative potential but greater evidence of maturation with the most mature endothelial cells displaying no proliferative potential. In preliminary studies, human umbilical cord blood was enriched in EPC compared to adult blood. We hypothesize that the frequency and distribution of EPC changes during ontogeny such that the number of circulating cells and EPC residing in vessels decline with age and there is loss of proliferative potential that correlates with telomerase activity. In other preliminary data, we have determined that cord blood EPC co- cultured with human marrow stem cells expands HSC >260 fold compared to freshly isolated marrow HSC. We now hypothesize that EPCwith the highest proliferative potential will better promote HSC expansion than EPC with lower proliferative potential. To test these hypotheses, we proposed the following aims: Specific Aim 1. Determine the role of human cord blood EPC in expanding human cord blood HSC ex vivo. Human cord blood will be co-cultured with EPC ¿ growth factors and the repopulating ability compared following injection into rhesus fetal hosts in utero. Specific Aim 2. Determine the frequency and proliferative potential of EPC derived from blood and vascular endothelium throughout rhesus ontogeny. Aortic endothelial cells and peripheral blood of rhesus subjects from fetal to aged adults will be examined for EPC content, proliferative potential, and telomerase activity. We hypothesize that EPC proliferative potential declines with age and the rhesus model permits us an opportunity to sample vessels and blood throughout the entire species ontogeny. Specific Aim 3. Determine the role of EPC derived from fetal versus adult blood vessels in the in vitro expansion of fetal rhesus cord blood and adult marrow HSC. We hypothesize that the fetal EPC will more optimally expand fetal than adult HSC but adult HSC will be better expanded on fetal than adult EPC. Indiana University, Indianapolis, IN PHS 398 (Rev. 09/04) Page 98 Form Page 2 Principal Investigator/Program Director (Last, First, Middle): Tarantal, Alice F (Project 1) KEY PERSONNEL. See instructions. Use continuation pages as needed to provide the required information in the format shown below. Start with Principal Investigator. List all other key personnel in alphabetical order, last name first. Name eRA Commons User Name Organization Role on Project Yoder, Men/inC. Indiana Unversity Project Leader Ingram, David A. Indiana University Co-Investigator OTHER SIGNIFICANT CONTRIBUTORS Name Organization Role on Project Human Embryonic Stem Cells [3 No Q Yes If the proposed project Involves human embryonic stem cells, list below the registration number of the specific cell line(s) from the following list: http://stemcells.nih.aov/reqistrv/index.asp. Usecontinuation pages as needed. If a specificline cannot be referenced at this time, include a statement that one from the Registry will be used. Cell Line Disclosure Permission Statement. Applicable toSBIR/STTR Only. See instructions. I I Yes I I No PHS 398 (Rev. 09/04) Page 99 Form Page 2-continued Number the followingpages consecutively throughout the application. Do not use suffixes such as 4a, 4b. Principal Investigator/Program Director (Last, First, Middle): Tarantal, Alice F. (Project 1) DETAILED BUDGET FOR INITIAL BUDGET PERIOD FROM THROUGH DIRECT COSTS ONLY 09/01/05 08/31/06 PERSONNEL (Applicant organization only) % DOLLAR AMOUNT REQUESTED (omit cents) TYPE EFFORT INST. ROLE ON APPT. ON BASE SALARY FRINGE NAME PROJECT (months) PROJ. SALARY REQUESTED BENEFITS TOTAL Principal Mervin C. Yoder Investigator 12 10.0 180,000 18,000 3 227 21,227 David Ingram Co-l 12 5.0 0 0 0 Jennifer Stewart Res Tech 12 100.0 39,140 39,140 12 263 51,403 SUBTOTALS 57,140 15,490 72,6301 ^ CONSULTANT COSTS None 0 EQUIPMENT (Itemize} None 0 SUPPLIES (Itemize by category) ILJ Laboratory supplies 37,500 TRAVEL None PATIENT CARE COSTS INPATIENT OUTPATIENT ALTERATIONS AND RENOVATIONS (Itemize by category) OTHER EXPENSES (Itemize by category) Wells Center Research Core Use Fee $3,576 3,576 CONSORTIUM/CONTRACTUAL COSTS DIRECT COSTS SUBTOTAL DIRECT COSTS FOR INITIAL BUDGET PERIOD (Item 7a, Face Page) $ 113,706 CONSORTIUM/CONTRACTUAL COSTS FACILITIES AND ADMINISTRATIVE COSTS 58,559 TOTAL DIRECT COSTS FOR INITIAL BUDGET PERIOD $ 172,265 SBIR/STTR Only: FEE REQUESTED PHS 398 (Rev. 09/04) Page 100 Form Page 4

人类脐带血已被证明是一种有效的造血干细胞来源 (HSC)用于临床移植。成人更广泛使用这种材料的一个主要限制 患者的问题是脐带血样本中的HSC数量可能不足以移植到成年患者身上， 尤其是不相关和不匹配的收件人。脐带血中HSC扩增方法的确定 将是这一领域的重大进步。我们最近鉴定了一系列内皮祖细胞 (EPC)在血液中循环，但也包括人类内皮细胞内膜的一部分 恒河猴的血管。与造血系统类似，EPC以层次结构组织在 增殖能力最强的祖先会产生大量增殖潜力较小的后代，但 更多的成熟证据表明，最成熟的内皮细胞没有表现出增殖潜力。在……里面 初步研究表明，与成人血相比，人脐血中富含EPC。我们 假设EPC的频率和分布在个体发育过程中发生变化，使得EPC的数量 驻留在血管中的循环细胞和内皮祖细胞随着年龄的增长而减少，并且失去了增殖能力 与端粒酶活性相关。在其他初步数据中，我们已经确定脐带血EPC协同- 与新鲜分离的骨髓HSC相比，用人骨髓干细胞培养的HSC扩增260倍。 我们现在假设，具有最高增殖潜力的EPC将更好地促进HSC的扩张 而EPC的增殖能力较低。为了验证这些假设，我们提出了以下目标： 具体目的1.确定人脐血内皮祖细胞体外扩增人脐血HSC的作用。 人脐血与EPC生长因子共培养及再生能力比较 注射到子宫内的恒河猴胎儿宿主体内。特定目标2.确定频率和增殖 来源于血液和血管内皮细胞的EPC在恒河猴个体发育过程中的潜能。主动脉 将对恒河猴受试者从胎儿到老年人的内皮细胞和外周血进行EPC检查 内容物、增殖潜能和端粒酶活性。我们假设EPC具有增殖潜能 随着年龄的增长而下降，恒河猴模型允许我们有机会在整个过程中采样血管和血液 整个物种的个体发育。具体目标3.确定胎儿和成人血液来源的EPC的作用 胎儿脐血和成人骨髓HSC体外扩增中的血管。我们假设 胎儿EPC对胎儿HSC的扩增效果优于成人HSC，但成人HSC对胎儿HSC的扩增效果更好 而不是成年的EPC。 印第安纳大学，印第安纳波利斯，In 小灵通398(09/04版)第98页表格第2页 首席调查员/项目主任(最后、第一、中间)：Tarantal，Alice F(项目1) 关键人员。请参阅说明。根据需要使用续页，以如下所示的格式提供所需信息。 从首席调查员开始。按字母顺序列出所有其他关键人员，姓氏在前。 名称时代共享用户名项目上的组织角色 约德，曼斯/公司印第安纳大学项目负责人 大卫·A·英格拉姆印第安纳大学共同研究员 其他重要贡献者 命名项目中的组织角色 人类胚胎干细胞[3否Q是 如果建议的项目涉及人类胚胎干细胞，请从下面的列表中列出特定细胞系(S)的注册号： Http://stemcells.nih.aov/reqistrv/index.asp.根据需要使用二次评估页面。 如果此时不能引用某一特定行，请附上一项声明，说明将使用登记处的某一行。 细胞系 披露许可声明。仅适用于SBIR/STTR。请参阅说明。我我是我我不是 小灵通398(09/04版)第99页表格第2页-续 在整个申请过程中，请将以下页面连续编号。不要使用诸如4a、4b之类的后缀。 首席调查员/项目主任(最后、第一、中间)：Tarantal，Alice F.(项目1) 初始预算期间的详细预算，从 直接成本仅为09/01/05 08/31/06 人员(仅适用于申请组织)申请金额百分比(省略美分) 键入Effort Inst. 在APPT中的角色。关于基本工资和福利 项目名称(月)项目。薪资申请福利合计 本金 默文·C·约德调查员12 10.0 180,000 18,000 3 227 21,227 大卫·英格拉姆联席L 12 5.0 0 0 珍妮弗·斯图尔特科技公司12 100.0 39,140 39,140 12 263 51,403 小计57,140 15,490 72,6301 ^ 顾问费 无%0 设备(分项) 无 0 用品(按类别分项列出) ILJ实验室用品 37500 旅行 无 住院病人护理费用 门诊病人 改建和翻新(按类别分列) 其他费用(按类别分项列出) 富国银行研究中心核心使用费3,576美元 3576 联合体/合同成本直接成本 初步预算期的直接费用小计(项目7a，正面页)113,706美元 财团/合同费用、设施和行政费用58,559 初期预算期间的直接费用总额172,265美元 仅限SBIR/STTR：需要收费 小灵通398(09/04版)第100页表格第4页