Lentiviral Gene Therapy of X-Linked Agammaglobulinemia
Lentiviral Gene Therapy of X-Linked Agammaglobulinemia
批准号:
8458516
负责人:
David J Rawlings
金额:
$55.34万
依托单位国家:
美国
项目类别:
财政年份:
2004
资助国家:
美国
项目状态:
已结题
起止时间:
2004-08-01 至 2016-03-31
关键词:
AddressAffectAgammaglobulinaemia tyrosine kinaseAllelesAntigensAutologousB-Cell DevelopmentB-LymphocytesBacterial InfectionsBiological AssayBone MarrowCell LineageCell physiologyCellsChromatinClinicalClinical ProtocolsClinical TrialsCodon NucleotidesComplementary DNADataDevelopmentDiseaseElementsEnhancersEvolutionExhibitsFutureGene ExpressionGene TransferGenerationsGenesGenetic RecombinationGoalsHematopoietic stem cellsHistopathologyHumanImmune System DiseasesImmune responseImmunoglobulinsImmunologic Deficiency SyndromesImmunologicsIn VitroLaboratoriesLeadLentivirus VectorLife ExpectancyMarrowMediatingModelingMolecular ProfilingMusMutagenesisMyelogenousMyeloid CellsPatientsQuality of lifeRare DiseasesReceptor SignalingReceptors, Antigen, B-CellRecoveryRecurrenceRegimenRelative (related person)Retroviral VectorRiskRoleSafetyStaining methodStainsStem cellsTestingToll-like receptorsTransgenesTransplantationVertebral columnViralWorkX-Linked Agammaglobulinemiabaseconditioningcongenital immunodeficiencydesigngene therapygenotoxicityhuman diseaseimprovedin vivonovelpressureprogenitorpromoterreceptorreconstitutionresponserestorationvector
中文摘要
描述(由申请方提供):X连锁无丙种球蛋白血症(XLA)由布鲁顿酪氨酸激酶(Btk)功能缺陷引起,特征为早期B细胞发育严重阻滞。表达野生型Btk的B细胞在体内表现出很强的选择性优势;这表明将正常Btk cDNA引入自体造血干细胞(HSC)中可能导致XLA中的长期免疫重建。在以前的工作中,我们证明了γ-逆转录病毒和B谱系特异性的慢病毒(LV)Btk基因治疗可以导致Btk依赖性的拯救,B谱系的发展和功能在体内。值得注意的是,除了其在B细胞中的作用之外,Btk还通过在骨髓细胞上表达的多种受体被激活,这意味着Btk在宿主免疫应答中具有更广泛的作用。为了优化Btk转基因在B和髓系细胞中的表达谱,我们开发了一种新的LV平台,该平台使用内源性Btk启动子(Btkp)与两种替代元件[IgH链增强子Em或普遍存在的染色质开放元件(UCOE)元件]。使用这些LV来表达密码子优化的人Btk,我们观察到Btk依赖性B谱系发育和功能的拯救;以及骨髓细胞中Toll样受体(TLR)信号传导的恢复。总之,这些数据有力地支持了我们的目标,即在未来的XLA患者基因治疗试验中使用基于Btkp的LV。然而,在将这种方法应用于患者之前,仍有几个关键问题有待解决。首先,我们需要对EmBtkp与UCOE. BtkpLV在鼠XLA和来源于XLA患者的原代干细胞中的安全性和有效性进行全面的体内比较。其次,我们需要详细评估这些候选LV的潜在遗传毒性。因此,该提议被设计为检验以下假设:(1)利用内源性Btk启动子的LV将介导持续的、时间上适当水平的Btk基因表达,并在鼠和人细胞中拯救B和骨髓功能;(2)使用非清髓性骨髓预处理方案也将实现有效的Btk拯救,该方案密切模拟XLA中的未来临床方法;和(3)这些LV将显示很少或没有载体介导的诱变的风险。
英文摘要
DESCRIPTION (provided by applicant): X-linked agammaglobulinemia (XLA) results from deficient function of Bruton's tyrosine kinase (Btk) and is characterized by a severe block in early B-cell development. B cells that express wildtype Btk exhibit a strong selective advantage in vivo; suggesting that introduction of a normal Btk cDNA into autologous hematopoietic stem cells (HSC) may lead to long-term immunologic reconstitution in XLA. In previous work, we demonstrated that both gamma-retroviral and B lineage-specific, lentiviral (LV) Btk gene therapy can lead to rescue of Btk-dependent, B lineage development and function in vivo. Notably, in addition to its role in B cells, Btk is activated via multiple receptors expressed on myeloid cells implying a broader role for Btk in host immune responses. To optimize the expression profile of the Btk transgene in both B and myeloid lineage cells, we developed a novel LV platform using the endogenous Btk promoter (Btkp) in association with two alternative elements [the IgH chain enhancer Em or a ubiquitous chromatin opening element (UCOE) element]. Using these LV to express codon-optimized human Btk, we observed rescue of Btk-dependent, B-lineage development and function; and restoration of Toll-like receptor (TLR) signaling in myeloid cells. Together, these data strongly support our goal of using a Btkp-based LV in a future gene therapy trial for patients with XLA. Several key issues; however, remain to be addressed before this approach can be moved forward into patients. First, we need a comprehensive in vivo comparison of the safety and efficacy of EmBtkp vs. UCOE.Btkp LV in both murine XLA and primary stem cells derived from XLA patients. Second, we need a detailed assessment of potential genotoxicity of these candidate LV. Accordingly, this proposal is designed to test the hypotheses that: (1) LV utilizing the endogenous Btk promoter will mediate sustained, temporally appropriate levels of Btk gene expression and rescue of B and myeloid function in murine and human cells; (2) Efficient Btk rescue will also be achieved using a non-myeloablative marrow conditioning regimen that closely mimics a future clinical approach in XLA; and (3)These LVs will exhibit little or no risk of vector- mediated mutagenesis.
期刊论文(0)
专著(0)
科研奖励(0)
会议论文
An integrated strategy to define the functional and synergistic impact of T1D causal variants
-
批准号:9227381
-
项目类别:
-
资助金额:$441.14万
-
财政年份:2016
-
负责人:David J Rawlings
-
依托单位:
Lentiviral Gene Therapy of X-Linked Agammaglobulinemia
-
批准号:8825754
-
项目类别:
-
资助金额:$8.78万
-
财政年份:2014
-
负责人:David J Rawlings
-
依托单位:
Administrative Core
-
批准号:8278876
-
项目类别:
-
资助金额:$6.3万
-
财政年份:2012
-
负责人:David J Rawlings
-
依托单位:
Pre-clinical Modeling of Foamy Viral gene Therapy for Murine and Human SCID-X1
-
批准号:8278864
-
项目类别:
-
资助金额:$25.01万
-
财政年份:2012
-
负责人:David J Rawlings
-
依托单位:
B cell function and phenotype as predictors of therapeutic response to rituximab
-
批准号:8044994
-
项目类别:
-
资助金额:$39.36万
-
财政年份:2010
-
负责人:David J Rawlings
-
依托单位:
Lentiviral Gene Therapy for Wiskott-Aldrich Syndrome
-
批准号:7576150
-
项目类别:
-
资助金额:$99.39万
-
财政年份:2008
-
负责人:David J Rawlings
-
依托单位:
Lentiviral Gene Therapy for Wiskott-Aldrich Syndrome
-
批准号:7463332
-
项目类别:
-
资助金额:$28.6万
-
财政年份:2008
-
负责人:David J Rawlings
-
依托单位:
Lentiviral Gene Therapy for Wiskott-Aldrich Syndrome
-
批准号:8228037
-
项目类别:
-
资助金额:$71.55万
-
财政年份:2008
-
负责人:David J Rawlings
-
依托单位:
Lentiviral Gene Therapy for Wiskott-Aldrich Syndrome
-
批准号:7772315
-
项目类别:
-
资助金额:$80.36万
-
财政年份:2008
-
负责人:David J Rawlings
-
依托单位:
Lentiviral Gene Therapy for Wiskott-Aldrich Syndrome
-
批准号:8029506
-
项目类别:
-
资助金额:$71.55万
-
财政年份:2008
-
负责人:David J Rawlings
-
依托单位:
Interdisciplinary Training in Genome Engineering (Component 10 of 11)TL1
-
批准号:7503426
-
项目类别:
-
资助金额:$4.95万
-
财政年份:2007
-
负责人:David J Rawlings
-
依托单位:
Interdisciplinary Training in Genome Engineering (Component 10 of 11)TL1
-
批准号:7903234
-
项目类别:
-
资助金额:$10.28万
-
财政年份:2007
-
负责人:David J Rawlings
-
依托单位:
Core--Immune Function Studies
-
批准号:7478454
-
项目类别:
-
资助金额:$9.66万
-
财政年份:2007
-
负责人:David J Rawlings
-
依托单位:
Cell and Virus CORE
-
批准号:7600770
-
项目类别:
-
资助金额:$17.5万
-
财政年份:2007
-
负责人:David J Rawlings
-
依托单位:
Interdisciplinary Training in Genome Engineering(Component 10 of 11) RL9
-
批准号:7687345
-
项目类别:
-
资助金额:$16.56万
-
财政年份:2007
-
负责人:David J Rawlings
-
依托单位:
Interdisciplinary Training in Genome Engineering
-
批准号:7466773
-
项目类别:
-
资助金额:$4.95万
-
财政年份:2007
-
负责人:David J Rawlings
-
依托单位:
Gene Repair in Murine Hematopoietic Stem Cells (Component 6 of 11)
-
批准号:7503424
-
项目类别:
-
资助金额:$45.5万
-
财政年份:2007
-
负责人:David J Rawlings
-
依托单位:
Cell and Virus CORE
-
批准号:7903236
-
项目类别:
-
资助金额:$82.21万
-
财政年份:2007
-
负责人:David J Rawlings
-
依托单位:
Interdisciplinary Training in Genome Engineering (Component 10 of 11)TL1
-
批准号:7687346
-
项目类别:
-
资助金额:$5.17万
-
财政年份:2007
-
负责人:David J Rawlings
-
依托单位:
Gene Repair in Murine Hematopoietic Stem Cells (Component 6 of 11)
-
批准号:7661520
-
项目类别:
-
资助金额:$45.5万
-
财政年份:2007
-
负责人:David J Rawlings
-
依托单位:
海外基金