HSCT WITH TRANSDUCED HEMATOPOIETIC STEM CELLS

转导造血干细胞的 HSCT

• 批准号: 6598166
• 负责人: Robertson Parkman
• 金额: $ 18.41万
• 依托单位: UNIVERSITY OF SOUTHERN CALIFORNIA
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-06-05 至 2003-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6598166
• 关键词: CD34 molecule; CD38 molecule; Lentivirus; T cell receptor; acute lymphocytic leukemia; autologous transplantation; flow cytometry; hematopoietic stem cells; human subject; human therapy evaluation; neoplasm /cancer immunology; neoplasm /cancer immunotherapy; patient oriented research; pediatric neoplasm /cancer; polymerase chain reaction; radiation immunosuppression; stem cell transplantation

APPLICANT'S DESCRIPTION: The goal of this proposal is to evaluate the transplantation of positively selected autologous hematopoietic stem cells (HSC) in pediatric patients with acute lymphoblastic leukemia (ALL). Previous results with autologous HSC transplantation (HSCT) for ALL have reported an unacceptable high relapse rates possibly due to leukemia cells contaminating the transplanted HSC. HSC will be positively selected by fluorescence activated cell sorting (FACS) based upon immunophenotypic differences between HSC (CD34+, CD38-, yc-) and ALL cells (CD34+, CD38+, yc+). Positively selected HSC will be evaluated 1) for their hematopoietic potential and 2) the presence of residual leukemia cells by PCR analysis for leukemia specific clonotypic TCR-6 rearrangements. PCR analysis of the clonospecific TCR-6 rearrangement permits an independent assessment of the purity of the isolated HSC (Specific Aim 1). All patients will be prepared for HSCT with total body irradiation and VP-16, and their rate of lymphohematopoietic reconstitution determined following the transplantation of the positively selected HSC (Specific Aim 2). After the demonstration that positively selected HSC can engraft in a clinically relevant time frame, an aliquot of the isolated HSC will be transduced with a glucocerebrosidase (GC) containing lentivirus based vector prior to cryopreservation. Both the transduced and non-transduced HSC will then be transplanted. Lentivirus based vectors will be used since our previous work has shown that murine leukemia based vectors poorly transduced quiescent HSC. Using inverse PCR techniques the clonality and the multi- lineage progeny of the transduced HSC will be longitudinally assessed. If patients relapse, their relapsed leukemia cells will be isolated and assess for the presence of the transduced vector to determine if the transplanted HSC contributed to relapse (Specific Aim 3). Overall this grant is an attempt to demonstrate that the transplantation of positively selected HSC (Cb34+, CD38-, yc-) can successfully lymphohematopoietic reconstitute pediatric ALL patients and that lentivirus vectors can transduce pluripotent HSC.

申请人的描述：该提案的目的是评估 正面选择自体造血干细胞的移植 （HSC）在急性淋巴细胞白血病（全部）的小儿患者中。以前的 自体HSC移植（HSCT）的结果都报告了 不可接受的高复发率可能是由于白血病细胞污染的 移植的HSC。 HSC将通过荧光积极选择 基于免疫表型差异激活的细胞分选（FACS） HSC（CD34+，CD38-，YC-）和所有细胞（CD34+，CD38+，YC+）。积极选择 HSC将评估1）其造血潜力和2）存在 残留白血病细胞通过PCR分析用于白血病特异性clonotypic TCR-6重排。 Clonompific TCR-6重排的PCR分析 允许对分离的HSC的纯度进行独立评估（特定 目标1）。 所有患者将准备用于全身照射的HSCT 和VP-16，以及确定的淋巴瘤症的重建率 在移植正面选择的HSC之后（特定目标2）。 演示了积极选择的HSC可以在 临床上相关的时间范围，孤立的HSC的等分试样将是 用含有基于慢病毒的载体的葡萄糖脑溴糖苷酶（GC）转导的 在冷冻保存之前。 转导的和未转导的HSC都将 然后移植。 自从我们的 先前的工作表明，基于鼠白血病的载体转导较差 静止的HSC。 使用逆PCR技术克隆性和多数 转导HSC的谱系后代将进行纵向评估。 如果 患者复发，他们的复发性白血病细胞将被隔离并评估 对于转导载体的存在，以确定移植的HSC是否 有助于复发（特定目标3）。 总的来说，这笔赠款是一种尝试 证明正面选择的HSC的移植（CB34+，CD38-，， yc-）可以成功地淋巴肿瘤的重构儿科所有患者 慢病毒载体可以转导多能HSC。