Targeting dendritic cells for selective modulation of GVHD

靶向树突状细胞选择性调节 GVHD

基本信息

批准号：
8349468
负责人：
Terry Fry
金额：
$ 24.89万
依托单位：
DIVISION OF BASIC SCIENCES - NCI
依托单位国家：
美国
项目类别：
财政年份：
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/8349468
关键词：
Allogenic Antibodies Antigen Targeting Antigen-Presenting Cells Antigens B-Lymphocytes Biological Preservation Blood Bone Marrow Bone Marrow Transplantation CD19 gene Cells Characteristics Clinic Competence Complication Dendritic Cells Development Diabetes Mellitus Disease Resistance Donor Lymphocyte Infusion Drug usage Frequencies Functional disorder Genes Goals ITGAX gene Immune Immune response Immune system Immunologic Deficiency Syndromes Immunologics Immunosuppression Immunosuppressive Agents Immunotherapeutic agent Infection Interferon Type II Interleukin-10 Laboratories Malignant Neoplasms Mediating Modality Modeling Monoclonal Antibodies Morbidity - disease rate Mouse Strains Muramidase Mus Myeloid Cells Organ Pathway interactions Patients Phagocytes Pharmaceutical Preparations Phenotype Photopheresis Play Population Population Biology Production Protocols documentation Publishing Reaction Regimen Relapse Reporting Risk Role S100A8 gene S100A9 gene STAT1 gene Severities Severity of illness Signal Transduction Staging Systemic Therapy T cell response T-Cell Depletion T-Lymphocyte Tissues Translating Treatment Efficacy Tumor Antigens Vaccination Vaccines Withdrawal Work base cancer cell cytokine exenatide graft vs host disease graft vs leukemia effect in vitro testing in vivo inflammatory modulation inhibitor/antagonist interferon gamma receptor interferon gamma receptors leukemia mortality novel pathogen prevent promoter recombinase reconstitution research study response transcription factor tumor

项目摘要

The experiments conducted under aim 1 have demonstrated that even relatively mild GVHD can diminish quantitative T cell immune responses to vaccination and functional immune responses to tumors expressing vaccine-targeted antigens. This work has been published (Capitini et al, Blood, 2009) and demonstrated the importance of preventing GVHD if BMT is to be optimized as a platform for immunotherapeutic approaches targeting malignancy. Under aim 2, we have established that inhibition of interferon gamma signaling can prevent the development of GVHD (Capitini et al, Blood 2009). Reduction in GVHD severity with disruption of interferon gamma signaling on T cells was not surprising given the known importance of this cytokine in GVHD but this approach results in immunodeficiency. The novel finding in these studies was that selective loss of interferon gamma receptor on bone marrow-derived non-T cells also prevented GVHD mediated by T cells with intact interferon gamma signaling and did so with preservation of qualitative and functional responses to vaccines. We have also established the extracorporeal photopheresis, a modality currently being used in the clinic to treat GVHD, also prevented GVHD with preserved vaccine response via modulation of IL-10 production in DC populations (Capitini et al, Biology of Blood and Marrow Transplantation, 2011). We next studied whether other components of the interferon gamma pathway could be targeted in donor bone marrow to prevent GVHD. Using bone marrow deficient in STAT1, a transcription factor necessary for interferon gamma signaling, we have confirmed that interference with this pathway in bone marrow-derived cells can prevent GVHD with preserved immune competence. To identify the relevant bone-marrow-derived cell population, we have selectively targeted STAT1 by generating mice with a floxed STAT1 gene (obtained from Dr. Lothar Hennighausen) that express the Cre recombinase under non-T cell promoters (CD11c (DC expression), lysozyme (on all phagocytic cells), and CD19 (B cell expression). In recipients of bone marrow from these donors, the STAT1 gene (and, thus, interferon gamma signaling) can be ablated in selective cell populations. Selective loss of STAT1 in all of these cell populations was not sufficient to prevent GVHD. Interestingly, further assessment of DC reconstitution in recipients of allogeneic STAT1 deficient bone marrow demonstrated expanded plasmacytoid DC (pDC) populations. Further more, we have confirmed that STAT1 remained intact in all of the Cre floxed STAT1 mouse strains generated thus far. We have obtained pDC-depleting antibodies (from Dr. Giorgio Trinchieri) to confirm that the expanded pDC populations are responsible for the GVHD protection observed in recipients of STAT1-deficinet bone marrow. Preliminary studies indicate that this is case. Based on these findings we have begun to analyze the characteristics of the expanded pDCs mediating GVHD resistance. Preliminary studies indicate that there is an increase in the frequency of CD9 negative pDCs, reported to be tolerogenic. In addition, expression of S100A8 and S100A9, genes associated with a suppressive phenotype in myeloid cells, is increased in pDCs from STAT1 deficient bone marrow recipients. Using S100A8/S100A9 deficient mice (obtained from Dr. Dimitri Gabrilovich through the Mackall laboratory) we have confirmed that loss of these genes in donor bone marrow increases the severity of GVHD. Aim 3 is in the initial stage. We have obtained selective Stat1 inhibitors and are beginning to test these in vitro. In addition, we used off-target inhibition of STAT1 by exenatide, a drug used in patients with diabetes, to demonstrate reduction of GVHD severity in our murine models.

根据AIM 1进行的实验表明，即使是相对温和的GVHD也可以减少对疫苗接种的定量T细胞免疫反应以及对表达疫苗靶向抗原抗原的肿瘤的功能免疫反应。这项工作已发表（Capitini等，Blood，2009），并证明了如果要优化BMT作为针对恶性肿瘤的免疫治疗方法的平台，则预防GVHD的重要性。在AIM 2下，我们确定抑制干扰素伽马信号传导可以防止GVHD的发展（Capitini等，Blood 2009）。鉴于GVHD中该细胞因子的重要性，T细胞上的干扰素伽马信号传导的破坏降低了GVHD的严重程度，但这种方法导致免疫缺陷。在这些研究中的新发现是，在骨髓来源的非T细胞上选择性丧失了干扰素伽马受体在骨髓衍生的非T细胞上还阻止了由具有完整干扰素伽马信号传导的T细胞介导的GVHD，并在保留了对疫苗的定性和功能反应的过程中这样做。我们还建立了体外光遗化，这是一种目前在诊所中用于治疗GVHD的方式，还通过调节DC种群中的IL-10产生来防止GVHD通过保留的疫苗反应（Capitini等人，血液和骨髓移植生物学，2011年）。接下来，我们研究了干扰素伽马途径的其他组件是否可以针对供体骨髓以防止GVHD。使用STAT1缺陷的骨髓（干扰素伽马信号传导所必需的转录因子），我们已经证实，在骨髓衍生的细胞中对该途径的干扰可以防止具有保留的免疫能力的GVHD。为了识别相关的骨髓衍生的细胞群，我们通过用floxed Stat1基因（从Lothar Hennighausen博士获得的小鼠）进行选择性靶向STAT1，该基因从Lothar Hennighausen博士获得了非T细胞启动子下的CRE重组酶（CD11C（DC表达）（DC表达），Lysozyme，lysozyme，lysozyme（在所有phagococycy cellist cyperors cypys cypys cypys cys）中（BORNISTIC）（bone）和CD19（bone）（bone）（bone）（bone）（bone）（bone）（bone）（bone）（bone）。 STAT1基因（因此，干扰素的信号传导）可以在所有这些细胞种群中的选择性损失中散发出来。到目前为止，我们已经获得了pdc的抗体（从Giorgio trinchieri博士那里）确认PDC种群的GVHD保护是在STAT1-DEFICIFICINET bone的特征中，因此，CRE的STAT1小鼠菌株（从Giorgio trinchieri博士那里得到了GVHD的范围。介导GVHD的耐药性。 Dimitri Gabrilovich通过Mackall实验室，我们已经确认供体骨髓中这些基因的损失会增加GVHD的严重性。我们的鼠模型中的严重程度。