TY - JOUR
T1 - PTD-mediated intracellular delivery of mutant NFAT minimum DNA binding domain inhibited the proliferation of T cells
AU - Liu, Xia
AU - Zhao, Qianqian
AU - Peng, Xin
AU - Xia, Sheng
AU - Shen, Weihong
AU - Zong, Yangyong
AU - Cheng, Jing
AU - Wu, Weijiang
AU - Zhang, Miaomiao
AU - Du, Fengyi
AU - Xu, Wenrong
AU - Qian, Hui
AU - Shao, Qixiang
N1 - Funding Information:
This work was supported by the following grants: The National Natural Science Foundation of China (grant numbers: 30671984, 81273202, 31200676 and 81172834), The Natural Science Foundation of Jiangsu Province of China (grant number: BK2008231), Clinical Medicine Science & Technology Project of Jiangsu Province of China (grant number: BL2013024), Program of Innovative Research Team of Jiangsu Province China, Construct Program of the Key Discipline of Jiangsu Province and Construct Program of the Key Discipline of Jiangsu University. The Postgraduate Students' Innovation Program of General Higher Education of Jiangsu Province of China (grant numbers: CX10B_283Z, CXZZ11_0591, CXLX12_0675).
PY - 2014/3
Y1 - 2014/3
N2 - The nuclear factor of activated T cell (NFAT) family of calcium-regulated transcription factors plays a key role in the development and function of the immune system. Calcineurin, a protein phosphatase, activates NFAT by dephosphorylation. The activated NFAT is translocated into the nucleus, where it up-regulates the expression of interleukin 2 (IL-2) and other target genes. Calcineurin inhibitors such as cyclosporine A (CsA) and FK506 are effective immunosuppressant drugs and dramatically increase the success rate of organ transplantation procedures. However, since calcineurin is expressed in most tissues in the body and calcineurin inhibition alters many cellular processes besides immune cell activation, the therapeutic use of calcineurin inhibitors is limited by serious side effects. Thus inhibiting NFAT by other mechanisms such as blocking its binding to DNA could be a more selective and safer approach to target NFAT for therapeutic applications. In peripheral T cells, productive immune responses are dependent upon the cooperative binding of the NFAT/AP-1 transcriptional complex to the promoter regions of genes such as interleukin-2 (IL-2), while NFAT in the absence of AP-1 leads to T cell anergy. Protein transduction domains (PTDs) are able to penetrate cell membranes and can be used to transport exogenous proteins across the cell and nuclear membranes. In this study, we constructed a fusion protein of PTD and a minimum DNA binding domain of human NFAT1 (PTD-ΔNFATminiDBD), which contains two mutations (R466A and T533G) in the AP-1 binding sites. The delivery and functions of this fusion protein in T cells were investigated. The results indicated that PTD-ΔNFATminiDBD could be effectively delivered into T cells and transported into the nucleus. PTD-ΔNFATminiDBD attenuated IL-2 production in T cells and then inhibited T cell proliferation, likely through competing against endogenous NFAT for binding to the IL-2 gene promoter. These results demonstrated that PTD-ΔNFATminiDBD was an effective NFAT inhibitor with a novel mechanism of action and might potentially be used as an immunosuppressant for organ transplantation with higher safety and better tolerance than calcineurin inhibitors.
AB - The nuclear factor of activated T cell (NFAT) family of calcium-regulated transcription factors plays a key role in the development and function of the immune system. Calcineurin, a protein phosphatase, activates NFAT by dephosphorylation. The activated NFAT is translocated into the nucleus, where it up-regulates the expression of interleukin 2 (IL-2) and other target genes. Calcineurin inhibitors such as cyclosporine A (CsA) and FK506 are effective immunosuppressant drugs and dramatically increase the success rate of organ transplantation procedures. However, since calcineurin is expressed in most tissues in the body and calcineurin inhibition alters many cellular processes besides immune cell activation, the therapeutic use of calcineurin inhibitors is limited by serious side effects. Thus inhibiting NFAT by other mechanisms such as blocking its binding to DNA could be a more selective and safer approach to target NFAT for therapeutic applications. In peripheral T cells, productive immune responses are dependent upon the cooperative binding of the NFAT/AP-1 transcriptional complex to the promoter regions of genes such as interleukin-2 (IL-2), while NFAT in the absence of AP-1 leads to T cell anergy. Protein transduction domains (PTDs) are able to penetrate cell membranes and can be used to transport exogenous proteins across the cell and nuclear membranes. In this study, we constructed a fusion protein of PTD and a minimum DNA binding domain of human NFAT1 (PTD-ΔNFATminiDBD), which contains two mutations (R466A and T533G) in the AP-1 binding sites. The delivery and functions of this fusion protein in T cells were investigated. The results indicated that PTD-ΔNFATminiDBD could be effectively delivered into T cells and transported into the nucleus. PTD-ΔNFATminiDBD attenuated IL-2 production in T cells and then inhibited T cell proliferation, likely through competing against endogenous NFAT for binding to the IL-2 gene promoter. These results demonstrated that PTD-ΔNFATminiDBD was an effective NFAT inhibitor with a novel mechanism of action and might potentially be used as an immunosuppressant for organ transplantation with higher safety and better tolerance than calcineurin inhibitors.
KW - IL-2
KW - Immunosuppressant
KW - Mutant minimum DNA binding domain of NFAT1
KW - NFAT
KW - PTD
UR - http://www.scopus.com/inward/record.url?scp=84893588571&partnerID=8YFLogxK
U2 - 10.1016/j.intimp.2014.01.001
DO - 10.1016/j.intimp.2014.01.001
M3 - Article
C2 - 24440534
AN - SCOPUS:84893588571
SN - 1567-5769
VL - 19
SP - 110
EP - 118
JO - International Immunopharmacology
JF - International Immunopharmacology
IS - 1
ER -