The E3 ligase HECTD4 regulates COX-2-dependent tumor progression and metastasis

J Joanna A. Vuille (Krantz Family Center for Cancer Research, Massachusetts General Hospital Cancer Center and Harvard Medical School) C Cem Tanriover (Krantz Family Center for Cancer Research, Massachusetts General Hospital Cancer Center and Harvard Medical School) E Ezgi Antmen (Krantz Family Center for Cancer Research, Massachusetts General Hospital Cancer Center and Harvard Medical School) D Douglas S. Micalizzi (Krantz Family Center for Cancer Research, Massachusetts General Hospital Cancer Center and Harvard Medical School) R Richard Y. Ebright S Sambhavi Animesh (Krantz Family Center for Cancer Research, Massachusetts General Hospital Cancer Center and Harvard Medical School) R Robert Morris (Krantz Family Center for Cancer Research, Massachusetts General Hospital Cancer Center and Harvard Medical School) S Soroush Hajizadeh (Krantz Family Center for Cancer Research, Massachusetts General Hospital Cancer Center and Harvard Medical School) Z Zachary J. Nicholson (Krantz Family Center for Cancer Research, Massachusetts General Hospital Cancer Center and Harvard Medical School) H Hunter C. Russell (Krantz Family Center for Cancer Research, Massachusetts General Hospital Cancer Center and Harvard Medical School) E Eric F. Zaniewski (Krantz Family Center for Cancer Research, Massachusetts General Hospital Cancer Center and Harvard Medical School) B Ben S. Wittner B Ben K. Wesley (Krantz Family Center for Cancer Research, Massachusetts General Hospital Cancer Center and Harvard Medical School) J Ji Eun Kwak (Krantz Family Center for Cancer Research, Massachusetts General Hospital Cancer Center and Harvard Medical School) J Julian Grunewald R Regan N. Szalay (Center for Computational and Integrative Biology, Massachusetts General Hospital) D Douglas B. Fox (Krantz Family Center for Cancer Research, Massachusetts General Hospital Cancer Center and Harvard Medical School) M Min Yang J J. Keith Joung (Krantz Family Center for Cancer Research, Massachusetts General Hospital Cancer Center and Harvard Medical School) D Doga C. Gulhan A Andrew E. H. Elia (Krantz Family Center for Cancer Research, Massachusetts General Hospital Cancer Center and Harvard Medical School) W Wilhelm Haas E Eugene Oh (Krantz Family Center for Cancer Research, Massachusetts General Hospital Cancer Center and Harvard Medical School) S Shyamala Maheswaran D Daniel A. Haber

Abstract

E3 ubiquitin ligases mediating turnover of proteins engaged in cancer progression point to key regulatory nodes. To uncover modifiers of metastatic competency, we conducted an in vivo genome-wide CRISPR-inactivation screen using cultured breast circulating tumor cells, following intravascular seeding and lung colonization. We identified HECTD4, a previously uncharacterized gene encoding a conserved potential homologous to E6AP C-terminus domain–containing ubiquitin transferase, as a potent tumor and metastasis suppressor. We show that purified HECTD4 mediates ubiquitin conjugation in vitro, and proteomic studies combined with ubiquitin remnant profiling identify a major degradation target as the prostaglandin synthetic enzyme cyclooxygenase-2 ( COX-2 ; PTGS2 ). In addition to COX-2 itself, HECTD4 targets its regulatory kinase MKK7. In breast cancer models, HECTD4 expression is induced as cells lose adherence to the matrix, and its depletion massively increases COX-2 expression, enhancing anchorage-independent proliferation and tumorigenesis. Genetic or pharmacologic suppression of COX-2 reverses the protumorigenic and prometastatic phenotype of HECTD4-depleted cells. Thus, HECTD4 encodes an E3 ubiquitin ligase that downregulates COX-2 suppressing anchorage independence in epithelial cancer cells.

Article Details

Volume / Issue Vol. 122, Issue 32
Published August 12, 2025
ISSN 0027-8424
Publisher National Academy of Sciences

Authors (25)

J

Joanna A. Vuille

Krantz Family Center for Cancer Research, Massachusetts General Hospital Cancer Center and Harvard Medical School

C

Cem Tanriover

Krantz Family Center for Cancer Research, Massachusetts General Hospital Cancer Center and Harvard Medical School

E

Ezgi Antmen

Krantz Family Center for Cancer Research, Massachusetts General Hospital Cancer Center and Harvard Medical School

D

Douglas S. Micalizzi

Krantz Family Center for Cancer Research, Massachusetts General Hospital Cancer Center and Harvard Medical School

R

Richard Y. Ebright

S

Sambhavi Animesh

Krantz Family Center for Cancer Research, Massachusetts General Hospital Cancer Center and Harvard Medical School

R

Robert Morris

Krantz Family Center for Cancer Research, Massachusetts General Hospital Cancer Center and Harvard Medical School

S

Soroush Hajizadeh

Krantz Family Center for Cancer Research, Massachusetts General Hospital Cancer Center and Harvard Medical School

Z

Zachary J. Nicholson

Krantz Family Center for Cancer Research, Massachusetts General Hospital Cancer Center and Harvard Medical School

H

Hunter C. Russell

Krantz Family Center for Cancer Research, Massachusetts General Hospital Cancer Center and Harvard Medical School

E

Eric F. Zaniewski

Krantz Family Center for Cancer Research, Massachusetts General Hospital Cancer Center and Harvard Medical School

B

Ben S. Wittner

B

Ben K. Wesley

Krantz Family Center for Cancer Research, Massachusetts General Hospital Cancer Center and Harvard Medical School

J

Ji Eun Kwak

Krantz Family Center for Cancer Research, Massachusetts General Hospital Cancer Center and Harvard Medical School

J

Julian Grunewald

R

Regan N. Szalay

Center for Computational and Integrative Biology, Massachusetts General Hospital

D

Douglas B. Fox

Krantz Family Center for Cancer Research, Massachusetts General Hospital Cancer Center and Harvard Medical School

M

Min Yang

J

J. Keith Joung

Krantz Family Center for Cancer Research, Massachusetts General Hospital Cancer Center and Harvard Medical School

D

Doga C. Gulhan

A

Andrew E. H. Elia

Krantz Family Center for Cancer Research, Massachusetts General Hospital Cancer Center and Harvard Medical School

W

Wilhelm Haas

E

Eugene Oh

Krantz Family Center for Cancer Research, Massachusetts General Hospital Cancer Center and Harvard Medical School

S

Shyamala Maheswaran

D

Daniel A. Haber