Characterizing the Mechanism of DPP8/9 Inhibitor-Induced Pyroptosis

DPP8/9 抑制剂诱导细胞焦亡机制的表征

• 批准号: 10733874
• 负责人: Daniel Bachovchin
• 金额: $ 52.38万
• 依托单位: SLOAN-KETTERING INST CAN RESEARCH
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-02-23 至 2028-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10733874
• 关键词: Acceleration; Affect; Amino Acid Sequence; Amino Acids; Area; Autoimmune Diseases; Biology; C-terminal; Cancer Etiology; Cell Death; Cells; Cellular Metabolic Process; Cellular Structures; Communicable Diseases; Complex; Cysteine; Data; Dipeptides; Dipeptidyl Peptidases; Disease; Foundations; Future; Health; Homeostasis; Human; Immune response; Immunity; Inflammasome; Inflammatory; Innate Immune System; Knowledge; Laboratories; Length; Ligand Binding; Link; Lytic; Malignant Neoplasms; Mass Spectrum Analysis; Mediating; Metabolic; Metabolic Diseases; Metabolic dysfunction; Mitochondria; Molecular; Monitor; Multiprotein Complexes; N-terminal; Natural Immunity; Neurodegenerative Disorders; Oxidation-Reduction; Paper; Pattern recognition receptor; Peptides; Play; Printing; Process; Proline; Protein Region; Proteins; Proteomics; Public Health; Reactive Oxygen Species; Reporting; Repression; Research; Research Project Grants; Serine; Signal Transduction; Stress; T-Lymphocyte; Therapeutic; Work; disulfide bond; human disease; inhibitor; multicatalytic endopeptidase complex; pathogen; peptidomimetics; protein degradation; receptor binding; small molecule

PROJECT SUMMARY Several human pattern-recognition receptors detect intracellular danger-associated signals, oligomerize into multiprotein complexes called inflammasomes, and trigger a lytic form of cell death called pyroptosis. Inflammasomes are involved in mounting immune responses to pathogens and in maintaining organismal homeostasis, but their hyperactivation can cause cancer, autoimmune disorders, and metabolic dysfunction. As such, it is critically important to characterize the molecular mechanisms that regulate inflammasome activation. NLRP1 and CARD8 are related pattern-recognition receptors that form inflammasomes, but the danger signals that they sense have not been fully established. Notably, ligands that bind to the serine dipeptidyl peptidases 8 and 9 (DPP8/9), including endogenous peptides with Xaa-Pro (where Xaa is any amino acid) N-termini, have been reported to activate these inflammasomes. However, why the innate immune system monitors Xaa-Pro peptide levels is unknown and constitutes a major knowledge gap. Recently, reductive stress, or a profound lack of reactive oxygen species (ROS), was also reported to activate the NLPR1 and CARD8 inflammasomes. The central hypothesis of this application is that reductive stress and Xaa-Pro peptide accumulation are intimately related danger signals that together comprise an overall “danger state” that causes rapid and full NLRP1 and CARD8 inflammasome activation. Specifically, it is proposed that the disordered regions of many cytosolic proteins, including the autoinhibitory N-terminal region of CARD8, are stabilized by intramolecular disulfide bonds; reductive stress abolishes these bonds, destabilizing these sequences and triggering their degradation into peptides by the proteasome. Proline is the most abundant amino acid in disordered protein regions, and therefore reductive stress likely generates many Xaa-Pro peptides. In this way, Xaa-Pro peptide accumulation can serve to confirm that reductive stress is occurring. This central hypothesis has been formulated based on preliminary data produced in the applicant’s laboratory and described in this application. The objective of this project is to determine the relationship between reductive stress, disordered protein degradation, and Xaa-Pro peptide accumulation. This project consists of three Specific Aims: 1) to determine how reductive stress induces the proteasome-mediated degradation of CARD8, 2) to characterize the relationship between reductive stress and Xaa-Pro peptide accumulation, and 3) to determine the relationship between cell metabolism and inflammasome activation. The successful completion of this work will not only clarify the primordial function of these enigmatic inflammasomes, but will also reveal a previously unknown a connection between intracellular redox state and protein stability. Moreover, this work will provide the foundation for future efforts to therapeutically control these inflammasomes for the treatment of human disease.

项目摘要 几种人类模式识别受体检测细胞内与信号转导相关的信号，寡聚化成 多蛋白复合物称为炎性小体，并触发细胞死亡的裂解形式称为pyroptosis。 炎性小体参与对病原体的免疫反应， 它们的过度激活可导致癌症、自身免疫性疾病和代谢功能障碍。作为 因此，表征调节炎性小体活化的分子机制是至关重要的。 NLRP 1和CARD 8是形成炎性小体的相关模式识别受体，但危险信号 他们认为还没有完全建立起来。值得注意的是，与丝氨酸二肽基肽酶8结合的配体 和9（DPP 8/9），包括具有Xaa-Pro（其中Xaa是任何氨基酸）N-末端的内源肽，具有 据报道可以激活这些炎性小体。然而，为什么先天免疫系统监测Xaa-Pro 肽水平是未知的，构成了一个主要的知识缺口。最近，还原压力，或一个深刻的 还报道了缺乏活性氧（ROS）激活NLPR 1和CARD 8炎性体。 本申请的中心假设是还原性应激和Xaa-Pro肽积累是导致细胞凋亡的重要因素。 密切相关的危险信号，共同构成了整体的“危险状态”， NLRP 1和CARD 8炎性小体激活。具体地说，它提出，许多无序区域 细胞溶质蛋白，包括CARD 8的自抑制性N-末端区域，通过分子内抑制剂稳定。 二硫键;还原应激消除这些键，使这些序列不稳定并触发它们的 被蛋白酶体降解为肽。脯氨酸是无序蛋白质中含量最丰富的氨基酸 区域，因此还原应激可能产生许多Xaa-Pro肽。通过这种方式，Xaa-Pro肽 积累可用于确认还原应激正在发生。这一核心假设一直被 基于在申请人的实验室中产生并在本申请中描述的初步数据配制。 本项目的目的是确定还原性应激、蛋白质紊乱之间的关系， 降解和Xaa-Pro肽积累。该项目包括三个具体目标：1）确定 还原性应激如何诱导蛋白酶体介导的CARD降解8，2）以表征 还原胁迫和Xaa-Pro肽积累之间的关系，以及3）确定 细胞代谢和炎性小体激活之间的联系这项工作的顺利完成不仅将 阐明了这些神秘的炎性小体的原始功能，但也将揭示一个以前未知的， 细胞内氧化还原状态和蛋白质稳定性之间的联系。此外，这项工作将提供 为将来努力治疗性控制这些炎性小体以治疗人类疾病奠定了基础。

项目成果

Protein folding stress potentiates NLRP1 and CARD8 inflammasome activation.

• DOI: 10.1016/j.celrep.2022.111965
• 发表时间: 2023-01
• 期刊: Cell reports
• 影响因子: 8.8
• 作者: Elizabeth L. Orth-He;Hsin-Che Huang;Sahana D. Rao;Qinghui Wang;Qifeng Chen;Claire M. O’Mara;Ashley J. Chui;Michelle Saoi;A. R. Griswold;A. Bhattacharjee;D. Ball;J. Cross;D. Bachovchin

DPP8/9 are not Required to Cleave Most Proline-Containing Peptides.

切割大多数含脯氨酸的肽不需要 DPP8/9。

• DOI: 10.1002/ijch.202200117
• 发表时间: 2023
• 期刊: Israel journal of chemistry
• 影响因子: 3.2
• 作者: Bhattacharjee,Abir;Bachovchin,DanielA
• 通讯作者: Bachovchin,DanielA

Activation of the CARD8 Inflammasome Requires a Disordered Region.

• DOI: 10.1016/j.celrep.2020.108264
• 发表时间: 2020-10-13
• 期刊: Cell reports
• 影响因子: 8.8
• 作者: Chui AJ;Griswold AR;Taabazuing CY;Orth EL;Gai K;Rao SD;Ball DP;Hsiao JC;Bachovchin DA
• 通讯作者: Bachovchin DA

The NLRP1 and CARD8 inflammasomes detect reductive stress.

NLRP1和CARD8炎症体检测还原应力。

• DOI: 10.1016/j.celrep.2022.111966
• 发表时间: 2023-01-31
• 期刊: Cell reports
• 影响因子: 8.8

DPP8/DPP9 inhibitor-induced pyroptosis for treatment of acute myeloid leukemia.

• DOI: 10.1038/s41591-018-0082-y
• 发表时间: 2018-08
• 期刊: Nature medicine
• 影响因子: 82.9
• 作者: Johnson DC;Taabazuing CY;Okondo MC;Chui AJ;Rao SD;Brown FC;Reed C;Peguero E;de Stanchina E;Kentsis A;Bachovchin DA
• 通讯作者: Bachovchin DA