Anti-Tumor Activity of Camptothecin Analog Conjugate of an RSPO4-Based Peptibody Targeting LGR4/5/6 in Preclinical Models of Colorectal Cancer

Authors

Yukimatsu Toh
Ling Wu
Jianghua Tu
Zhengdong Liang
Adela M Aldana
Jake J Wen
Li Li
Sheng Pan
Julie H Rowe
Martha E Hensel
Carolyn L Hodo
Rick A Finch
Kendra S Carmon
Qingyun J Liu

Language

English

Publication Date

8-18-2025

Journal

British Journal of Cancer

DOI

10.1038/s41416-025-03121-2

PMID

40825849

PMCID

PMC12478594

PubMedCentral® Posted Date

9-30-2025

PubMedCentral® Full Text Version

Author MSS

Abstract

Background: Antibody-drug conjugates (ADCs) are a significant advancement in targeted cancer therapy, but none are approved for colorectal cancer (CRC). LGR4/5/6, highly expressed in most CRCs, are promising targets. While LGR5-targeting ADCs show strong anti-tumor effects, their efficacy is limited by LGR5 loss in some CRC cells. RSPO4, a natural ligand for LGR4/5/6, binds all three receptors with high affinity. This research develops RSPO4-based peptibody drug-conjugates (PDCs) to simultaneously target LGR4/5/6, offering a novel therapeutic approach for CRC.

Methods: LGR4/5/6 expression in CRCs was analysed using RNA-seq datasets and Western blot. Peptibody binding affinities were measured, conjugated to camptothecin analog, CPT2, and tested for cytotoxicity in CRC cell lines. Antitumor efficacy was evaluated in vivo using CRC cell line and patient-derived xenograft (PDX) models.

Results: Peptibody was engineered by fusing a mutant RSPO4 furin-domain to human IgG1 Fc, retaining high-affinity LGR4/5/6 binding without enhancing Wnt/β-catenin signalling. Conjugated with CPT2 molecules, the PDC showed strong antitumor activity in CRC cell lines and dose-dependent tumor growth inhibition in xenograft and patient-derived models.

Conclusion: Preclinical data showed that LGR4/5/6-targeting PDC exhibited potent cytotoxicity in vitro and robust antitumor efficacy in CRC xenograft and PDX models, making its potential as a promising therapeutic approach for CRC.

Published Open-Access

yes

Recommended Citation

Toh, Yukimatsu; Wu, Ling; Tu, Jianghua; et al., "Anti-Tumor Activity of Camptothecin Analog Conjugate of an RSPO4-Based Peptibody Targeting LGR4/5/6 in Preclinical Models of Colorectal Cancer" (2025). The Brown Foundation: Institute of Molecular Medicine. 14.
https://digitalcommons.library.tmc.edu/molecular_med/14