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

> **NIH NIH F30** · WEILL MEDICAL COLL OF CORNELL UNIV · 2021 · $51,036

## Abstract

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.

## Key facts

- **NIH application ID:** 10200713
- **Project number:** 5F30CA243444-03
- **Recipient organization:** WEILL MEDICAL COLL OF CORNELL UNIV
- **Principal Investigator:** Andrew Griswold
- **Activity code:** F30 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2021
- **Award amount:** $51,036
- **Award type:** 5
- **Project period:** 2019-07-01 → 2023-06-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10200713

## Citation

> US National Institutes of Health, RePORTER application 10200713, Characterizing the DPP8/9 Pyroptotic Checkpoint with Chemoproteomic Protease Substrate Profiling (5F30CA243444-03). Retrieved via AI Analytics 2026-05-22 from https://api.ai-analytics.org/grant/nih/10200713. Licensed CC0.

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