Enhancement of Hematopietic Stem Cell Engraftment with CD+ Marrow Progenitors

这份指导临床科学家发展奖(K08)的建议书描述了5年培训计划 给马修·J·舒切特博士。这项建议建立在候选人的优势和先前的研究技能基础上 并利用匹兹堡大学研究科学家的指导和资源。 通过安格斯·汤姆森博士的指导，以及结构化的教学部分，候选人的 执行假设驱动的研究的能力将提高到完全独立的外科科学家。 该项目广泛的、长期的目标是评估CD8+骨髓-CD8+的机制(S)。 CD8+/TCR-亚群促进(促进)异基因造血干细胞植入 小鼠的障碍，导致多系融合和供体特异性移植的发展 没有GVHD的耐受性。最近的研究表明，类似于两种浆的细胞告诉pDC (CD8+/CD11c+/B220+/CD11b-)和发育中的T细胞(PTCs：CD8+/CD3+)存在于FC人群中， 它们各自不能达到完全的促进作用。因此，我们假设 CD8+PDC联合应用促进异基因造血干细胞植入 和PTC前体细胞协同工作以增强其耐受性。 具体目标1：评估单个CD8+/TCR-骨髓来源亚群的功能贡献。 -PDC和PTC亚群的联合给药将在体内进行评估，以检查促进潜力。 具体目标2：评估pDC和pTCs的细胞运输和分化模式。 -EGFP和荧光素酶标记的PDC和PTC将在同种异体移植后进行研究。 具体目标3：评价FC在体外和体内的作用机制。 -将评估与共刺激、细胞因子极性和Treg诱导相关的潜在机制 具体目标4：鉴定和表征人类骨髓中的易化亚群。 -将表征人骨髓中的CD8+/TCR-亚群，并评估NOD-SCID小鼠的促进潜力。 这些研究的数据将为诱导耐受的基于细胞的疗法提供重要的见解。 相关性(请参阅说明)： 更好地了解PDC和PTC的相互作用将有助于创新的纯化细胞的开发 血液系统恶性肿瘤的治疗和支持的组成部分治疗策略 人类干细胞在广泛的临床环境中的应用(血球蛋白疾病、免疫缺陷、组织 再生和缺血性心力衰竭)。这些知识将为胸部器官移植带来新的希望。 受者和患有各种先天性和获得性血液病的患者。

This mentored clinical scientist development award (K08) proposal describes the 5 year training program for Dr. Matthew J. Schuchert. This proposal builds upon the Candidate's strengths and prior research skills and takes advantage of the guidance and resources of the research scientists at the University of Pittsburgh. Through the mentorship of Dr. Angus Thomson, as well as a structured didactic component, the Candidate's ability to perform hypothesis-driven research will be advanced to a fully-independent surgeon scientist. The broad, long term objective of this project is to evaluate the mechanism(s) by which CD8+ marrow- derived subsets (CD8+/TCR-) enhance (facilitate) hematopoietic stem cell engraftment across allogeneic barriers In mice, resulting In the development of multllineage chlmerism and donor-specific transplantation tolerance without GVHD. Recent studies have demonstrated that cells resembling both plasmacytold pDCs (CD8+/CD11c+/B220+/CD11b-) and developing T cells (pTCs: CD8+/CD3+) exist within the FC population, which individually fail to achieve the complete facilitation effect. Therefore, we hypothesize that facilitation of allogeneic hematopoietic stem cell engraftment Is enhanced by the co-adminlstratlon of discrete CD8+ pDC and pTC progenitors working in concert to augment their tolerogenic effect. Specific Aim 1: Evaluate the functional contributions of individual CD8+/TCR- marrow-derived subsets. - Co-administration of pDC and pTC subsets will be assessed in vivo to examine facilitating potential. Specific Aim 2: Assess the cell trafficking and differentiation patterns of pDCs and pTCs. - EGFP and luclferase labelled pDC and pTC will be studied following allogeneic transplantation. Specific Aim 3: Evaluate potential mechanisms of FC function in vitro and in vivo. - Will evaluate potential mechanisms related to co-stimulation, cytokine polarity and Treg induction Specific Aim 4: Identification and characterization of facilitating subsets In human marrow. - Will characterize CD8+/TCR- subsets In human marrow, and assess facilitating potential in NOD-scid mice. Data from these studies will shed important Insights on cell-based therapies for tolerance induction. RELEVANCE (See instructions): A better understanding of pDC and pTC interactions will aid the development of innovative purified cell component therapeutic strategies for the treatment of hematopoietic malignancies, as well as for the support of human stem cells in a wide variety of clinical settings (hemaglobinopathies, immunodeficiencies, tissue regeneration, and ischemic heart failure). Such knowledge would afford new hope in thoracic organ transplant recipients and in patients suffering from a wide range of congenital and acquired hematologic disorders.