C/EBPβ Regulation of the Type 1 IFN Response; Sensitizing Keratinocytes to Direct Activators of Cytosolic PRRs and DNA Damage-Induced Cell Death

C/EBPβ 1 型干扰素反应的调节；

基本信息

批准号：
10735531
负责人：
Jonathan Russell Hall
金额：
$ 32.04万
依托单位：
NORTH CAROLINA STATE UNIVERSITY RALEIGH
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-07-15 至 2028-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10735531
关键词：
Antiviral Response Apoptosis Apoptotic Autoimmune Diseases B-DNA Biological CASP3 gene CASP8 gene CCAAT-Enhancer-Binding Protein-beta Cell Death Cell Death Induction Cellular Stress Cessation of life Communicable Diseases Cutaneous DNA DNA Damage Data Disease Double-Stranded RNA Epidermis Genes Genetically Engineered Mouse Immune Induction of Apoptosis Infection prevention Infectious Skin Diseases Inflammatory Innate Immune Response Interferon Type I Interferon-beta Interferons Laboratories Link Malignant Neoplasms Mediating Microbe Modification Molecular Mus Nerve Degeneration Nucleic Acids Outcome Pathway interactions Pattern recognition receptor Play Poly I-C Proliferating RNA Receptor Signaling Regulation Repression Research Role Skin Skin Cancer Up-Regulation Viral Virus Virus Diseases antimicrobial experimental study in vivo keratinocyte new therapeutic target novel pathogen receptor response senescence transcription factor type I interferon receptor

项目摘要

Cellular stress, DNA damage, pathogen insult, and immune mechanisms can activate molecular pathways that result in regulated cell death. Increasing our understanding of the pathways that regulate cell death decisions in response to cellular stress is critical as misregulated cell death is linked to cancer, neurodegeneration and autoimmune diseases. The epidermis is the first line of defense to cutaneous microbes, viruses and environmental insults, and keratinocytes play a critical role in the host innate immune response mediated by type I interferons (IFN-I). In addition to anti-microbial and anti-viral responses this same IFN-I response also mediates diverse cellular and biological responses such as proliferation, apoptosis, senescence and the DNA damage response. Recent data from out laboratory suggest that the CCAAT/enhancer-binding protein-β (C/EBPβ) transcription factor is a novel regulator the keratinocyte type IFN-I response. We observed the conditional deletion of C/EBPβ in mouse epidermis (CKOβ) resulted in increased expression of IFNb and numerous ISGs which included cytosolic pattern recognition receptors (cPRRs). cPRRs sense viral/pathogen RNA and DNA as well as damaged host cytosolic DNA and RNA to trigger a IFN-I response. CKOβ keratinocytes treated with direct activators of cytosolic PRRs displayed a greatly increased IFN-I response that surprisingly resulted in increased apoptosis via the activation of caspase-8 and caspase-3. As breifly mentioned, DNA damage can activate the innate immune response. DNA damaging agents can result in modifications and structural changes in RNA and DNA that could potentially be sensed by cytosolic PRRs. We observed that CKOβ keratinocytes challenged with DNA damage displayed increase caspase 8-mediated apoptosis that was dependent on the interferon a/b receptor (INFAR). Activation of a type IFN-I response can result in the increased expression of ISGs with apoptotic functions that activate caspase-8 mediated apoptosis via death receptor signaling. The objective of this proposal is to understand molecular basis for how C/EBPβ negatively regulates the IFN-I response and caspase-8 mediated apoptosis in response to direct activators of cytosolic PRRs and DNA damage in vivo in mouse epidermis and in primary keratinocytes. We hypothesize that C/EBPβ negatively regulates the IFN-I response in keratinocytes and the loss of C/EBPβ sensitizes keratinocytes to the direct activation of cytosolic PRRs by pathogen RNA/DNA or damaged host RNA/DNA to induce pro-apoptotic ISGs and caspase 8-mediated apoptosis. These experiments will increase our understanding of a novel pathway by which C/EBPβ regulates activation of cPRRs and the IFN-I response to control cell death decisions in response to cellular stress including DNA damage. This increased understanding could identify new therapeutic targets to restore proper regulation of cell death to treat skin cancer, skin infectious diseases, and skin autoimmune diseases.

细胞应激、DNA 损伤、病原体损伤和免疫机制可以激活分子途径，导致受调节的细胞死亡增加我们对调节细胞死亡决策的途径的了解。对细胞应激的反应至关重要，因为失调的细胞死亡与癌症、神经退行性变和表皮是皮肤微生物、病毒和自身免疫性疾病的第一道防线。环境损伤和角质形成细胞在宿主先天免疫反应中发挥着关键作用 I 型干扰素 (IFN-I) 除了抗微生物和抗病毒反应外，还具有相同的 IFN-I 反应。介导多种细胞和生物反应，如增殖、凋亡、衰老和 DNA 来自实验室损伤的最新数据表明 CCAAT/增强子结合蛋白-β (C/EBPβ) 转录因子是角质形成细胞 IFN-I 反应的新型调节因子。小鼠表皮中 C/EBPβ (CKOβ) 的条件性缺失导致 IFNb 和 IFNb 表达增加许多 ISG，其中包括胞质模式识别受体 (cPRR)，可感知病毒/病原体。 RNA 和 DNA 以及受损的宿主细胞质 DNA 和 RNA 来触发 IFN-I 反应。用胞质 PRR 直接激活剂处理的角质形成细胞显示出大大增强的 IFN-I 反应，令人惊讶的是，As breifly 通过激活 caspase-8 和 caspase-3 导致细胞凋亡增加。提到的，DNA损伤可以激活先天免疫反应，从而导致DNA损伤。 RNA 和 DNA 的修饰和结构变化可能会被胞质 PRR 感知。观察到受到 DNA 损伤的 CKOβ 角质形成细胞显示 caspase 8 介导的增加依赖于干扰素 a/b 受体 (INFAR) 的细胞凋亡可以激活 IFN-I 型反应。导致具有凋亡功能的 ISG 表达增加，激活 caspase-8 介导的细胞凋亡该提案的目的是了解 C/EBPβ 的分子基础。负调节 IFN-I 反应和 caspase-8 介导的细胞凋亡，以响应直接激活剂小鼠表皮和原代角质形成细胞体内的胞质 PRR 和 DNA 损伤。 C/EBPβ 负向调节角质形成细胞中的 IFN-I 反应，并且 C/EBPβ 的缺失会导致敏化角质形成细胞通过病原体 RNA/DNA 或受损的宿主 RNA/DNA 直接激活胞质 PRR 诱导促凋亡 ISG 和 caspase 8 介导的细胞凋亡。这些实验将增加我们的研究。了解 C/EBPβ 调节 cPRR 激活和 IFN-I 反应的新途径控制细胞死亡决策以应对细胞应激，包括 DNA 损伤。了解可以确定新的治疗靶点，以恢复细胞死亡的适当调节来治疗皮肤癌症、皮肤传染病、皮肤自身免疫性疾病。