The TRH-TRHR Axis Acts as a Metabolic Checkpoint for Ammonia Tolerance in AML: Therapeutic Targeting via Imiquimod and Itraconazole
Yan Wang, Xinhui Qiu, Weiwei Liu, Zhixiang Lei, Zheying Wang, Xiaoyu Zhang, Hongxing Wang, Hongchun WangIntroduction:
Metabolic reprogramming facilitates survival in Acute Myeloid Leukemia (AML), but the molecular checkpoints governing ammonia tolerance remain unclear. This study aimed to investigate the role of the TRH/TRHR axis in AML ammonia tolerance and identify potential therapeutic interventions.
Methods:
Ammonia Metabolism-Related Genes (AMRGs) were initially identified, and core prognostic genes were screened using 101 combinations of 10 machine learning algorithms. The optimal drug combination was derived by using the Biological Factor-Regulated Neural Network (BFReg- NN) model, followed by molecular docking. A risk signature was constructed and evaluated using survival analysis, time-dependent Receiver Operating Characteristic (ROC) curves, calibration curves, Decision Curve Analysis (DCA), and Cox regression. By systematically assessing ammonia levels, cell proliferation, apoptosis, and ROS accumulation, alongside functional rescue and genetic validation experiments, we elucidated the underlying mechanisms of ammonia tolerance in AML and validated the therapeutic efficacy of the screened drugs.
Results:
A total of 6 core prognostic genes (TRH, TCIRG1, CXCL10, RUFY4, GPR12, CCR5) were screened out through machine learning. The BFReg-NN model predicted imiquimod and itraconazole as the optimal drug combination. The constructed risk model demonstrated moderate prognostic predictive ability. An abnormal "high ammonia and low TRH" state in AML was identified. Exogenous TRH treatment significantly reversed ammonia tolerance, suppressed AML cell proliferation, increased ROS accumulation, and promoted AML cell death. Combination treatment with imiquimod and itraconazole significantly inhibited AML cell proliferation and promoted apoptosis, with TRH/TRHR signaling playing an important role. Additional validation in THP-1 cells and TRHR knockdown experiments further supported the involvement of TRH/TRHR signaling.
Discussion:
Collectively, AML cells demonstrate a state of high ammonia tolerance. Our findings suggest that the TRH-TRHR axis may serve as an ammonia metabolism-related checkpoint in AML and indicate that the imiquimod-itraconazole regimen represents a potential therapeutic strategy for targeting ammonia tolerance in AML.
conclusion:
Collectively, AML cells demonstrate a state of high ammonia tolerance. Our findings establish the TRH-TRHR axis as a critical Ammonia metabolic checkpoint and demonstrate the translational potential of the Imiquimod-Itraconazole regimen as a precision therapy for targeting ammonia tolerance in AML.
Conclusion:
AML cells exhibit a state of high ammonia tolerance associated with reduced TRH expression. Our study suggested that TRH/TRHR signaling is involved in regulating ammonia tolerance in AML.