Identification of the extracellular membrane protein ENPP3 as a major cGAMP hydrolase and innate immune checkpoint

Rachel Mardjuki; Songnan Wang; Jacqueline Carozza; Bahar Zirak; Vishvak Subramanyam; Gita Abhiraman; Xuchao Lyu; Hani Goodarzi; Lingyin Li

doi:10.1016/j.celrep.2024.114209

Identification of the extracellular membrane protein ENPP3 as a major cGAMP hydrolase and innate immune checkpoint

Cell Rep. 2024 May 28;43(5):114209. doi: 10.1016/j.celrep.2024.114209. Epub 2024 May 14.

Authors

Rachel Mardjuki¹, Songnan Wang², Jacqueline Carozza³, Bahar Zirak⁴, Vishvak Subramanyam⁴, Gita Abhiraman⁵, Xuchao Lyu⁶, Hani Goodarzi⁴, Lingyin Li⁷

Affiliations

¹ Department of Biochemistry, Stanford University, Stanford, CA 94305, USA; ChEM-H Institute, Stanford University, Stanford, CA 94305, USA; Arc Institute, Palo Alto, CA 94304, USA; Department of Chemistry, Stanford University, Stanford, CA 94305, USA.
² Department of Biochemistry, Stanford University, Stanford, CA 94305, USA; ChEM-H Institute, Stanford University, Stanford, CA 94305, USA; Arc Institute, Palo Alto, CA 94304, USA.
³ Arc Institute, Palo Alto, CA 94304, USA.
⁴ Arc Institute, Palo Alto, CA 94304, USA; Department of Urology, University of California, San Francisco, San Francisco, CA 94143, USA; Helen Diller Family Comprehensive Cancer Center, University of California, San Francisco, San Francisco, CA 94158, USA; Department of Biophysics & Biochemistry, University of California, San Francisco, San Francisco, CA 94143, USA; Baker Computational Health Science Institute, University of California, San Francisco, San Francisco, CA 94143, USA.
⁵ Department of Molecular and Cellular Physiology, Stanford University, Stanford, CA 94305, USA.
⁶ ChEM-H Institute, Stanford University, Stanford, CA 94305, USA; Department of Pathology, Stanford University, Stanford, CA 94305, USA.
⁷ Department of Biochemistry, Stanford University, Stanford, CA 94305, USA; ChEM-H Institute, Stanford University, Stanford, CA 94305, USA; Arc Institute, Palo Alto, CA 94304, USA. Electronic address: lingyin@arcinstitute.org.

PMID: 38749434
DOI: 10.1016/j.celrep.2024.114209

Abstract

2'3'-Cyclic guanosine monophosphate (GMP)-AMP (cGAMP) is a second messenger synthesized upon detection of cytosolic double-stranded DNA (dsDNA) and passed between cells to facilitate downstream immune signaling. Ectonucleotide pyrophosphatase phosphodiesterase I (ENPP1), an extracellular enzyme, was the only metazoan hydrolase known to regulate cGAMP levels to dampen anti-cancer immunity. Here, we uncover ENPP3 as the second and likely the only other metazoan cGAMP hydrolase under homeostatic conditions. ENPP3 has a tissue expression pattern distinct from ENPP1's and accounts for all cGAMP hydrolysis activity in ENPP1-deficient mice. Importantly, we also show that, as with ENPP1, selectively abolishing ENPP3's cGAMP hydrolysis activity results in diminished cancer growth and metastasis of certain tumor types in a stimulator of interferon genes (STING)-dependent manner. Both ENPP1 and ENPP3 are extracellular enzymes, suggesting the dominant role that extracellular cGAMP must play as a mediator of cell-cell innate immune communication. Our work demonstrates that ENPP1 and ENPP3 non-redundantly dampen extracellular cGAMP-STING signaling, pointing to ENPP3 as a target for cancer immunotherapy.

Keywords: CP: Immunology; ENPP1; ENPP3; PD-1; STING; breast cancer; cGAMP; clinical trial; hydrolase; immune checkpoint; melanoma.

MeSH terms

Animals
Humans
Hydrolysis
Immunity, Innate*
Membrane Proteins* / metabolism
Mice
Mice, Inbred C57BL
Neoplasms / genetics
Neoplasms / immunology
Neoplasms / metabolism
Neoplasms / pathology
Nucleotides, Cyclic* / metabolism
Phosphoric Diester Hydrolases* / genetics
Phosphoric Diester Hydrolases* / metabolism
Pyrophosphatases* / genetics
Pyrophosphatases* / metabolism
Signal Transduction

Substances

Nucleotides, Cyclic
cyclic guanosine monophosphate-adenosine monophosphate
Phosphoric Diester Hydrolases
Membrane Proteins
Pyrophosphatases
ectonucleotide pyrophosphatase phosphodiesterase 1
Sting1 protein, mouse
ENPP3 protein, human