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Metabolic Disruption Induced by mTOR Signaling Pathway Inhibition in Regulatory T-Cell Expansion for Clinical
Roberto Gedaly1,2,3,4, Gabriel Orozco1, Alexandre P Ancheta1,4
1Transplant Division, Department of Surgery, College of Medicine, University of Kentucky, Lexington, KY 40536, USA.
Background:
Regulatory T cell (Treg) therapy is considered an alternative approach to induce tolerance in transplantation. If successful, this therapy may have implications on immunosuppression minimization/withdrawal to reduce drug-induced toxicity in patients. The aim of this study was to assess the efficacy of the mTORC1/C2 inhibitor, AZD8055, in the manufacturing of clinically competent Treg cells and compare the effects with those induced by rapamycin (RAPA), another mTOR inhibitor commonly used in Treg expansion protocols.
Methods:
Primary human Treg cells were isolated from leukapheresis product. Cell viability, expansion rates, suppressive function, autophagy, mitochondrial unfolded protein response (mitoUPR), and cell metabolic profile were assessed.
Results:
We observed a stronger inhibition of the mTORC2 signaling pathway and downstream events triggered by Interleukin 2 (IL2)-receptor in AZD8055-treated cells compared with those treated with RAPA. AZD8055 induced progressive metabolic changes in mitochondrial respiration and glycolytic pathways that disrupted the long-term expansion and suppressive function of Tregs. Unlike RAPA, AZD8055 treatment impaired autophagy and enhanced the mitoUPR cell stress response pathway.
Conclusions:
A distinct pattern of mTOR inhibition by AZD, compared with RAPA, induced mitochondrial stress response and dysfunction, impaired autophagy, and disrupted cellular bioenergetics, resulting in the loss of proliferative potential and suppressive function of Treg cells.
Insights
AZD8055 impairs regulatory T cell (Treg) expansion and function by inducing mitochondrial stress and disrupting cellular metabolism, unlike rapamycin. This finding is crucial for Treg therapy development in transplantation.
Area of Science:
- Immunology
- Cell Biology
- Pharmacology
Background:
- Regulatory T cell (Treg) therapy offers a promising approach for inducing transplant tolerance.
- Minimizing immunosuppression through Treg therapy can reduce drug-related toxicity in patients.
Purpose of the Study:
- To evaluate the efficacy of AZD8055, an mTORC1/C2 inhibitor, in manufacturing clinical-grade Treg cells.
- To compare AZD8055's effects with rapamycin (RAPA) in Treg expansion protocols.
Main Methods:
- Human Treg cells were isolated and assessed for viability, expansion, suppressive function, autophagy, mitochondrial unfolded protein response (mitoUPR), and metabolic profile.
- Comparative analysis of AZD8055 and RAPA effects on Treg cells.
Main Results:
- AZD8055 strongly inhibited mTORC2 signaling and IL2-receptor downstream events more than RAPA.
- AZD8055 induced metabolic dysregulation in mitochondrial respiration and glycolysis, impairing Treg expansion and function.
- AZD8055 impaired autophagy and enhanced mitoUPR, unlike RAPA.
Conclusions:
- AZD8055 triggers mitochondrial stress, dysfunction, impaired autophagy, and bioenergetic disruption in Tregs.
- These effects lead to a loss of Treg proliferative potential and suppressive function, contrasting with RAPA's effects.
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