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

• 批准号: 8222979
• 负责人: Abdoreza Davoodi-Semiromi
• 金额: $ 21.29万
• 依托单位: UNIVERSITY OF FLORIDA
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-22 至 2012-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8222979
• 关键词: Generating; regulatory; cells; JAK; STAT5

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(申请人提供)：我们最近发现Tyrphostin AG490通过IL-2-JAK-Stat5信号通路阻断NOD和B6小鼠调节性T细胞中Stat5的磷酸化。我们还报道了在NOD小鼠中，Stat5B与DNA的结合亲和力较弱是由于该基因的DNA结合域发生了点突变，并证明该突变是NOD鼠特有的突变。在这些研究过程中，我们使用JAK激酶抑制剂Tyrphostin AG490来阻断自然调节性T细胞中由IL-2刺激的Stat5B的激活。令人震惊的是，在体外用AG490处理CD4？Foxp3-T细胞后，这个群体变成了CD4？包括NOD在内的几个不同品系小鼠的T细胞。我们的初步数据显示，CD4？？经AG490转化的T细胞对TCR刺激无反应，并抑制CD4？Foxp3-T细胞体外培养。与性别和年龄匹配的假治疗NOD对照组小鼠(n=9)相比，腹膜注射AG490显著延迟NOD小鼠(P&lt；0.004，n=8)的糖尿病发病。这是我们关于Tyrphostin AG490产生调节性T细胞的新发现的后续研究，我们提出了两个特定的目标；1)通过过继地将糖尿病脾细胞与转基因NOD.BDC2.5小鼠的CD4T细胞共同转移来预防/推迟1型糖尿病的发生，AG490在免疫缺陷的NOD.SCID小鼠中进行再编程，以及2)通过过继地将非再生性脾细胞与NOD胰岛抗原特异性T细胞重新编程相结合来预防/推迟1型糖尿病的发生。综上所述，我们报道了一种在体外获得大量调节性T细胞的新方法，并提出了预防NOD小鼠T1D的方法。与其他已发表的方法相比，我们用AG490产生调节性T细胞的方法成本低廉，而且非常快速。该方法可能提供一种在体外制造大量调节性T细胞的方法，可用于T1D预防和抑制同种异体胰岛移植排斥反应的细胞治疗。项目相关性-据我们所知，我们首次在小鼠体内描述了Tyrphostin AG490的免疫调节特性。通过酪氨酸蛋白AG490对常规T细胞重新编程来产生抗原特异的调节性T细胞是非常有意义的。我们关于Tyrphostin AG490产生调节性T细胞的发现是高度创新和新颖的，具有很大的潜力转化为临床，延迟/预防自身免疫性疾病和/或防止同种异体移植的排斥反应。我们的方法非常便宜，而且快速，可以在任何普通免疫学实验室进行。我们非常热衷于通过这项提案中描述的方法来改进调节性T细胞的制造，通过为这项拨款申请提供资金，我们将有机会产生令人信服的体外和体内数据，并达到适合临床前研究的水平。

项目成果

Correspondence regarding "Long term treatment with ACE inhibitor enalapril decreases body weight gain and increases life span in rats".

关于“长期使用 ACE 抑制剂依那普利治疗可降低大鼠体重增加并延长寿命”的通讯。

• DOI: 10.1016/j.bcp.2011.12.033
• 发表时间: 2012
• 期刊: Biochemical pharmacology
• 影响因子: 5.8
• 作者: Davoodi-Semiromi,Abdoreza;Cooper-DeHoff,Rhonda
• 通讯作者: Cooper-DeHoff,Rhonda

Influence of Tyrphostin AG490 on the expression of diabetes-associated markers in human adipocytes.

Tyrphostin AG490 对人类脂肪细胞中糖尿病相关标志物表达的影响。

• DOI: 10.1007/s00251-012-0659-4
• 发表时间: 2013
• 期刊: Immunogenetics
• 影响因子: 3.2
• 作者: Davoodi-Semiromi,Abdoreza;Wasserfall,CH;Hassanzadeh,A;Cooper-DeHoff,RM;Wabitsch,M;Atkinson,M
• 通讯作者: Atkinson,M