Molecular Basis of Familial Glucocorticoid Deficiency

2015 Fiscal Year Research-status Report

• Project/Area Number: 26461542
• Research Institution: Hamamatsu University School of Medicine
• Principal Investigator: 山口 理恵 浜松医科大学, 医学部, 特任研究員 (50420360)
• Project Period (FY): 2014-04-01 – 2017-03-31
• Keywords: NNT activity / patient / lymphocyte / bioenergetics / mitochondria

Outline of Annual Research Achievements

1. Effects of NNT knockdown on mitochondrial bioenergetics in HAC15 cells
Results: Both mitochondrial oxygen consumption and cellular ATP levels were greatly decreased in the siNNT-transfected cells compared to the control cells, suggesting that the mitochondrial bioenergetics is impaired by NNT knockdown.
2. Mitochondrial function and bioenergetics in the patient's lymphocytes
Results: The NNT activity was markedly lower in the patient compared to the controls. In keeping with the in vitro NNT knockdown results, the rate of ROS production in the lymphocytic mitochondria was higher and the mitochondrial-dependent oxygen consumption was diminished in the patient, implying that NNT deficiency impairs mitochondrial bioenergetics due to increased oxidative stress.

Current Status of Research Progress

4: Progress in research has been delayed.

Reason

Evaluation of mitochondrial function and bioenergetics in the patient's lymphocytes were not able to be carried out as smoothly as planned.

Strategy for Future Research Activity

1. Investigation of the effects of NNT defect on p38 MAPK signaling pathway
2. Investigation of the effects of NNT defect on JNK and apoptotic pathway
3. Identification of novel genes responsible for FGD and clarify the underlying mechanism(s)

Causes of Carryover

当初、研究計画していたもののうち、進行が遅れてしまったため、使用額に差が生じてしまった。

Expenditure Plan for Carryover Budget

siRNA、トランスフェクション試薬、real-time PCRやwestern blotting用試薬、 apoptosis detection kit、 antibodies against ERK/JNK/P38 MAPK、次世代シーケンシング用試薬などを購入予定である。