Generating regulatory T-cells by a JAK-STAT5 kinase inhibiotr: A noval approach t

通过 JAK-STAT5 激酶抑制剂生成调节性 T 细胞：一种新方法

基本信息

批准号：
7347457
负责人：
Abdoreza Davoodi-Semiromi
金额：
$ 18.13万
依托单位：
UNIVERSITY OF CENTRAL FLORIDA
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-09-22 至 2011-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7347457
关键词：
Affinity Age Allografting Antigens Applications Grants Autoimmune Diseases Bone Marrow CD4 Positive T Lymphocytes CD40 Antigens CD80 gene Cell Lineage Cell Therapy Cells Clinic Coculture Techniques Control Groups Cyclosporine DNA DNA Binding DNA Binding Domain Data Dendritic Cells Derivation procedure Diabetes Mellitus Diabetes prevention Dose Encephalomyelitis Family Family member Follow-Up Studies Funding Generations Genes Human IL2RA gene Immune system Immunization Immunology In Vitro Inbred NOD Mice Incidence Injection of therapeutic agent Insulin-Dependent Diabetes Mellitus Interleukin-2 JAK2 gene JAK3 gene Janus kinase Knowledge Laboratories Literature Lymphocyte Methods Molecular Molecular Weight Monitor Mouse Strains Mus Mutation Myelogenous Paralysed Pathway interactions Phenotype Phosphorylation Phosphotransferases Play Point Mutation Population Prevention Property Protein Tyrosine Kinase Publishing Rattus Regulation Reporting Role Signal Pathway Signal Transduction Specific qualifier value Splenocyte Summary Reports T-Cell Proliferation T-Lymphocyte Technology Transgenic Organisms Translating Transplantation Tyrosine Phosphorylation Tyrphostins base cell type computerized data processing heart allograft immunoregulation improved in vivo innovation insulin dependent diabetes mellitus onset interest islet islet allograft kinase inhibitor member novel novel strategies preclinical study prevent research study sex tyrphostin AG-490

项目摘要

DESCRIPTION (provided by applicant): We have recently shown that tyrphostin AG490 blocked phosphorylation of Stat5 via the IL-2-JAK-Stat5 signaling pathway in regulatory T-cells in NOD and B6 mice. We have also reported that weaker DNA binding affinity of Stat5B to the DNA in NOD mouse is due to a point mutation in DNA binding domain in this gene and demonstrated that this mutation is a unique mutation in NOD mouse. In the course of these studies, we used tyrphostin AG490, a JAK kinase inhibitor, to block activation of Stat5B by IL-2 stimulation in natural regulatory T- cells. Stunningly, in-vitro treatment of CD4?Foxp3- T-cells with AG490 converted this population into CD4? T-cells in several different mouse strains including NOD. Our preliminary data indicate that CD4?? T cells converted by AG490 are anergic to TCR stimulation and suppress proliferation of CD4? Foxp3- T cells in vitro. Interperitoneal injection of AG490 significantly delayed onset of diabetes in NOD mice (p<0.004, n=8) when compared with sex and age matched sham treated NOD control mice (n=9).This is a follow- up study of our novel finding regarding generating regulatory T-cells by tyrphostin AG490 and we propose two specific aims; 1) prevention/delay onset of type 1 diabetes by adoptive co-transfer of diabetogenic splenocytes with CD4 T-cells of transgenic NOD.BDC2.5 mice reprograms by AG490 in immunodeficient NOD.scid mice and, 2) prevention/delay onset of type 1 diabetes by adoptive co-transfer of diabetogenic splenocytes with NOD islet antigens-specific T-cell reprograms by AG490 in NOD.scid mice. In summary, we report a novel approach for the in- vitro derivation of large numbers of regulatory T-cells and propose to prevent T1D in NOD mouse. Our method to generate regulatory T-cells by AG490 is inexpensive and is very quick when compared with other published methods. This method may provide an approach for manufacturing a large number of regulatory T-cells in- vitro that can be used in cell-based therapies of T1D prevention and suppression of islet allograft rejection.Project relevance- To the best of our knowledge we are the first to describe immuno-regulatory properties of tyrphostin AG490 in mouse. Generating antigen-specific regulatory T-cells by reprogramming conventional T-cells by tyrphsotin AG490 is of great interest. Our finding regarding generating regulatory T-cells by tyrphostin AG490 is highly innovative and novel and has a great potential to translate into clinic and delay/prevent autoimmune diseases and/or to prevent rejection of allograft transplant. Our method is very inexpensive and is quick and can be performed in any general immunology laboratories. We have a great enthusiasm to improve manufacturing of regulatory T-cells by the method describe in this proposal and by funding this grant application we will have this opportunity to generate convincing in-vitro and in-vivo data and reach to a level suitable for a pre- clinical study.

描述（由申请人提供）：我们最近表明，在NOD和B6小鼠的调节性T细胞中，Tyrphostin AG490通过IL-2-JAK-STAT5信号通路阻断了STAT5的磷酸化。我们还报道说，NOD小鼠中Stat5b与DNA的DNA结合亲和力较弱，是由于该基因中DNA结合结构域的点突变引起的，并证明该突变是NOD小鼠的独特突变。在这些研究的过程中，我们使用了一种JAK激酶抑制剂Tyrphostin AG490通过自然调节T细胞中IL-2刺激来阻断STAT5B的激活。令人惊叹的是，用AG490的CD4？Foxp3-t细胞进行了体外处理将该人群转化为CD4？包括点头在内的几种不同小鼠菌株中的T细胞。我们的初步数据表明CD4？通过AG490转化的T细胞是厌食症的TCR刺激并抑制CD4的增殖吗？体外Foxp3-T细胞。与性别和年龄相匹配的假方法对照小鼠相比，腹膜间注射AG490显着延迟了NOD小鼠的糖尿病（p <0.004，n = 8）。 1）通过通过AG490在免疫缺陷的小鼠中通过AG490在AG490中对糖尿病生成的脾脏进行的预防/1型糖尿病的延迟开始。 iSlet抗原特异性T细胞重新编程在NOD.SCID小鼠中通过AG490。总而言之，我们报告了一种新型的方法，用于在体外衍生大量调节性T细胞，并提出预防NOD小鼠中T1D的方法。我们通过AG490生成调节性T细胞的方法很便宜，与其他已发表的方法相比，我们的方法非常快。该方法可以提供一种制造大量调节性T细胞体外的方法，该方法可用于预防T1D的基于细胞的疗法和抑制胰岛同种异体移植的抑制。对我们的最佳知识，我们是第一个描述小鼠中Tyrphostin Ag490的免疫调节性质的人。通过Tyrphsotin AG490重编程常规T细胞来产生抗原特异性的调节T细胞。我们关于泰弗蛋白AG490生成调节性T细胞的发现是高度创新性和新颖的，并且具有转化为临床和延迟/预防自身免疫性疾病和/或防止拒绝同种异体移植移植的巨大潜力。我们的方法非常便宜，并且可以在任何一般免疫学实验室中执行。通过本提案中描述的方法，我们非常热情地改善监管T细胞的制造，并通过资助本赠款申请，我们将有机会产生令人信服的视野和含量数据，并达到适合于临床前研究的水平。