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Characterizing the DPP8/9 Pyroptotic Checkpoint with Chemoproteomic Protease Substrate Profiling

使用化学蛋白质组蛋白酶底物分析表征 DPP8/9 焦亡检查点

基本信息

批准号：
10200713
负责人：
Andrew Griswold
金额：
$ 5.1万
依托单位：
WEILL MEDICAL COLL OF CORNELL UNIV
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-07-01 至 2023-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10200713
关键词：
Acute Myelocytic Leukemia Adult Benchmarking Biochemical Biological Biotechnology Biotin CASP9 gene CRISPR/Cas technology Caspase Cell Death Cell Line Cells Cellular Assay Chemicals Chemistry Cleaved cell Complex Coupling Data Diagnosis Dipeptidyl Peptidases Event Goals Growth Factor Human Immune response In Vitro Incidence Inflammasome Knock-out Label Lytic Malignant Neoplasms Mass Spectrum Analysis Methods Molecular Myeloid Cells N-terminal Oncology Organic Chemistry Outcome Pathway interactions Patients Peptide Hydrolases Peptides Pharmacology Physiological Physiological Processes Process Prognosis Proteasome Inhibitor Proteins Proteomics Recruitment Activity Research Resistance Serine Protease Techniques Technology Testing Therapeutic Tissues United States Work acute myeloid leukemia cell anticancer activity cancer initiation chemical group chemoproteomics cytokine cytotoxic inhibitor/antagonist innovation leukemia novel novel therapeutic intervention overexpression peptide I recruit response sensor small molecular inhibitor treatment strategy user-friendly

项目摘要

Project Summary/Abstract While acute myeloid leukemia (AML) is the most common form of leukemia in adults, prognosis remains poor. An emerging strategy for the treatment of AML centers around inhibiting the serine proteases DPP8 and DPP9 (DPP8/9), which activate the CARD8 inflammasome and triggers a lytic form of cell death known as pyroptosis. This process is directly cytotoxic to AML cells and indirectly activates the host immune response, delivering a dual mechanism for anticancer activity. Our central hypothesis is that DPP8/9 processing inactivates some as of yet unknown substrate(s) that, when active, recruit the inflammasome and triggers pyroptosis. Unfortunately, DPP8/9, like many of the ~550 human proteases, remain uncharacterized with respect their endogenous substrates. Identifying and characterizing the physiologically relevant substrates of DPP8/9 will be necessary to both understand and therapeutically exploit this pyroptotic checkpoint. Although proteases regulate countless important (patho)physiological processes, discovering protease substrates is extraordinarily challenging and state-of-the-art protease substrate profiling platforms are expensive and incomprehensive. In Specific Aim 1, I will advance a practical technology for protease substrate identification. This innovative platform uses an N-terminal reactive chemical moiety to selectively label the N-termini of proteins in complex cell lysates. Coupling this chemical group to biotin will enable the labeling, enrichment, and identification of endogenous protease substrates. In Specific Aim 2, I will use this technology, along with complimentary strategies to identify the substrates of DPP8/9, and then characterize how these substrates activate the CARD8 inflammasome. I anticipate that this project will lead to two key outcomes: 1) the introduction of a practical method for protease substrate discovery that will enable the unbiased interrogation of the many complex proteolytic networks that control important biological responses, and 2) the characterization of the DPP8/9 – CARD8 pyroptotic checkpoint, which has high potential to enable modulation of this pyroptotic pathway for the treatment of AML.

项目总结/文摘