Forty-eight hours later, I-E(52-68) peptide and subsequently either vehicle, RAPA or AZD2014 were injected i
Forty-eight hours later, I-E(52-68) peptide and subsequently either vehicle, RAPA or AZD2014 were injected i.p. IgG in allografts. Results The novel TORKinib AZD2014 impaired DC differentiation and T cell proliferation in vitro and depressed immune cells and allospecific T cell responses in vivo. A 9-day course of AZD2014 (10 mg/kg ip twice daily) or rapamycin (RAPA; 1mg/kg ip daily) prolonged median heart allograft survival time significantly (25 days for AZD2014; 100 days for RAPA; 9.5 days for control). Like RAPA, AZD2014 suppressed graft mononuclear cell infiltration, increased regulatory T cell (Treg) to effector memory T cell (Tem) ratios and reduced T follicular helper (Tfh) and B cells 7 days posttransplant. By 21 cIAP1 Ligand-Linker Conjugates 2 days (10 days after drug withdrawal), however, Tfh and B cells and donor-specific IgG1 and IgG2c antibody titers were significantly lower in RAPA- compared with AZD2014-treated mice. Elevated Treg to Tem ratios were maintained after RAPA, but not AZD2014 withdrawal. Conclusions Immunomodulatory effects of AZD2014, unlike those of RAPA, Mouse monoclonal to AXL were not sustained after drug withdrawal, possibly reflecting distinct pharmacokinetics or/and inhibitory effects of AZD2014 on mTORC2. Introduction The mechanistic target of rapamycin (mTOR) is an evolutionarily conserved serine/threonine kinase that drives organelle and cell growth1,2 through upregulation of glycolysis to fuel nucleotide, protein and lipid synthesis. mTOR functions as a component of at least 2 distinct multiprotein complexes,- mTOR complex 1 (mTORC1) and mTORC2.3,4 While both complexes include mLST8 (mammalian lethal with SEC13 protein 8) and DEPTOR cIAP1 Ligand-Linker Conjugates 2 (DEP domain-containing mTOR-interacting protein), mTORC1 uniquely associates with Raptor (regulatory associated protein of mTOR) and PRAS40 (proline-rich Akt substrate of 40 kDa). In contrast, mTORC2 associates with Rictor (rapamycin-insensitive companion of mTOR), mSIN1 (mammalian stress-activated protein kinase-interacting protein 1) and Protor (protein observed with cIAP1 Ligand-Linker Conjugates 2 Rictor). While mTORC1 has been implicated in regulation of nucleotide and protein synthesis, as well as autophagy, much less is known about the functions of mTORC2. Recently however, mTORC2 has been implicated in regulation of cell growth, proliferation, survival and cytoskeletal organization, as well as sodium handling in the kidney.5C7 There have been important recent advances in understanding of how mTOR complexes regulate immune cell differentiation and function.8 Thus, genetic deletion of either mTORC1 or mTORC2 in T cells has revealed that T helper (Th) 1 and Th17 differentiation is selectively regulated by mTORC1, whereas Th2 development is mTORC2-dependent.9,10 Furthermore, inhibition of both mTORC1 and mTORC2 favors regulatory T cell (Treg) development more than inhibition of either complex alone. In individual studies, small hairpin RNA vectors targeting Raptor (mTORC1) induce T follicular B helper cell (Tfh) differentiation at the expense of Th1 cells, while Rictor cIAP1 Ligand-Linker Conjugates 2 deletion promotes Th1 cells, with minimal effect cIAP1 Ligand-Linker Conjugates 2 on Tfh cells.11 While less is known concerning how mTOR impacts B cell function, deletion of Rictor in B cells causes marked deficiencies in mature follicular, marginal zone and B1a B cells with consequent effects on antibody (Ab) responses in vivo.12 The immunosuppressive prodrug rapamycin (RAPA) is an allosteric inhibitor of mTOR that mediates it effects indirectly via interaction with the immunophilin FK506 binding protein (FKBP) 12 and formation of a drug-immunophilin complex that directly binds the FKBP-rapamycin-binding (FRB) domain name of mTOR.13 While assembly of mTORC1 is RAPA-sensitive, mTORC2 is insensitive to RAPA. Recent studies in yeast demonstrating that this C terminal a part of Avo3, a subunit unique to mTORC2, prevents RAPA-FKBP12 from accessing the FRB domain name14 may help explain this phenomenon. To overcome shortcomings of RAPA and its analogues (rapalogs) as therapy for advanced malignancies, new generation adenosine triphosphate (ATP)-competitive mTOR inhibitors (TORKinibs) have been developed. By targeting both mTORC1 and mTORC2, these second generation mTOR inhibitors have been predicted to have more potent antitumor effects. Based on encouraging preclinical results, TORKinibs are being tested in early-phase clinical trials for treatment of advanced solid tumors or multiple myeloma.15,16 While much of our understanding of the effects of these TORKinibs stems from studies in oncology, little is known about their influence on immunity or their potential as immunosuppressive brokers. Recently, in a limited proof-of-principle study,17 we showed that this ATP-competitive mTOR inhibitor AZD8055 could suppress T cell proliferation and prolong.