Innate Immune Sensing as a Key Driver of Alcohol-Related Liver Injury = 알코올성 간질환을 유도하는 선천면역 감지 기전

다국어 초록 (Multilingual Abstract)

Alcohol consumption has short- and long-term impacts on physical and mental health. Although multiple host and environmental factors contribute to alcohol-related disease, the innate immune sensors that detect toxic signals from alcohol remain poorly defined. Here, we show that alcohol cooperates with interferon signaling to drive inflammatory cell death, cytokine release, and liver injury. We identified the pattern-recognition receptor ZBP1 as a key innate immune sensor mediating pyroptosis, apoptosis, and necroptosis in response to combined ethanol and interferon stimulation. While interferon elevated ZBP1, ethanol suppressed ADAR1 expression. In parallel, interferon and ethanol activated JNK signaling to promote Z-RNA formation, triggering ZBP1. These findings reveal a mechanism by which alcohol and interferon converge to induce ZBP1-dependent inflammatory cell death and liver pathology, providing mechanistic insight and highlighting potential therapeutic targets for alcohol-related disease.

국문 초록 (Abstract)

알코올 섭취는 신체적·정신적 건강 전반에 광범위한 영향을 미치며, 이에 따른 알코올 관련 질환은 전 세계적으로 중요한 보건 문제로 주목받고 있다. 알코올성 질환의 발생에는 다양한 숙주 요인과 환경 요인이 복합적으로 관여하지만, 알코올로 인해 유도되는 독성 신호를 감지하는 선천면역 센서는 아직 규명되지 않았다. 본 논문에서는 알코올이 인터페론 신호와 상호작용하여 염증성 세포사멸, 사이토카인 분비 및 간 손상을 유발함을 확인하였다. 이에 대한 자극을 매개하는 핵심 선천면역 센서로 ZBP1을 제시하고, 이는 파이롭토시스, 아포토시스 및 네크롭토시스를 매개한다. 한편, 인터페론 자극은 ZBP1의 발현을 증가시킨 반면, 에탄올은 RNA 편집 효소인 ADAR1의 발현을 억제하였다. 또한 두 자극은 c-Jun N-terminal kinase(JNK) 신호 전달 경로를 활성화하여 Z-RNA 형성을 촉진하였다. 이는 ZBP1 의존적인 염증성 세포사멸을 유도하여 알코올 관련 간 병리의 발생과 진행에 기여함을 보여준다. 종합하면, 본 연구는 알코올과 인터페론 신호가 선천면역 감지 기전을 통해 염증 반응과 간 손상을 매개함을 규명하였다.

목차 (Table of Contents)

• Abstract i
• Table of Contents iii
• List of Figures vi
• Chapter 1. Introduction 1
• 1.1. Study background 1
• 1.1.1. Alcoholic liver disease and inflammation 1
• 1.1.2. Interferon signaling 3
• 1.1.3. ZBP1-mediated inflammatory cell death 4
• 1.2. Purpose of research 5
• Chapter 2. Methods 7
• 2.1. Generation of immortalized BMDMs 7
• 2.2. Cell culture 7
• 2.3. Cell stimulation 8
• 2.4. Real-time analysis of cell death and lipid peroxidation 8
• 2.5. Immunoblot analysis 9
• 2.6. Dot blot analysis 11
• 2.7. Real-time quantitative PCR analysis 12
• 2.8. Mice 12
• 2.9. Immunofluorescence staining 13
• 2.10. In vivo ethanol feeding 14
• 2.11. Histopathology 15
• 2.12. ALT and cytokine analysis 15
• 2.13. RNA-sequencing and public data analysis 15
• 2.14. Statistical analysis 18
• Chapter 3. Results 19
• 3.1. Coupling of alcohol and inflammation triggers innate immune cell death 19
• 3.1.1. Alcohol and interferon induce cell death 19
• 3.1.2. Alcohol and interferon promote activation of pyroptosis, apoptosis, and necroptosis 25
• 3.1.3. Alcohol and interferon signaling trigger multiple lytic cell death pathways 30
• 3.2. Coupling of alcohol and interferon signaling exacerbates alcohol liver disease 33
• 3.2.1. Interferon signaling promotes alcohol-induced liver injury in vivo 33
• 3.2.2. Interferon signaling contributes to alcoholic liver disease in mice and is enriched in patients 37
• 3.2.3. Alcoholic liver disease is associated with multiple cell death pathways 40
• 3.3. ZBP1 is a sensor of ethanol and interferon 43
• 3.3.1. ZBP1 is upregulated in alcoholic liver disease and is required for liver injury in vivo 43
• 3.3.2. ZBP1 is required for alcohol- and interferon-induced inflammatory cell death 46
• 3.3.3. Alcohol- and interferon-induced cell death occurs independently of AIM2 and STING 49
• 3.4. Alcohol and interferon generate Z-RNA to activate ZBP1 51
• 3.4.1. Alcohol and interferon induce Z-RNA formation 51
• 3.4.2. Alcohol reduces ADAR1 to facilitate Z-RNA accumulation 54
• 3.4.3. JNK signaling promotes Z-RNA generation 56
• Chapter 4. Discussion 63
• Data Availability 66
• References 67
• Acknowledgements 74
• Abstract in Korean 75