Multiparatopic antibodies induce targeted downregulation of programmed death-ligand 1

Seth D Ludwig; Bunyarit Meksiriporn; Jiacheng Tan; Rakeeb Kureshi; Akhilesh Mishra; Kyle J Kaeo; Angela Zhu; Georgia Stavrakis; Stephen J Lee; David J Schodt; Michael J Wester; Dhiraj Kumar; Keith A Lidke; Andrea L Cox; Helen M Dooley; Sridhar Nimmagadda; Jamie B Spangler

doi:10.1016/j.chembiol.2024.02.014

Multiparatopic antibodies induce targeted downregulation of programmed death-ligand 1

Cell Chem Biol. 2024 May 16;31(5):904-919.e11. doi: 10.1016/j.chembiol.2024.02.014. Epub 2024 Mar 27.

Authors

Seth D Ludwig¹, Bunyarit Meksiriporn², Jiacheng Tan³, Rakeeb Kureshi³, Akhilesh Mishra⁴, Kyle J Kaeo¹, Angela Zhu¹, Georgia Stavrakis⁵, Stephen J Lee¹, David J Schodt⁶, Michael J Wester⁶, Dhiraj Kumar⁴, Keith A Lidke⁶, Andrea L Cox⁷, Helen M Dooley⁸, Sridhar Nimmagadda⁹, Jamie B Spangler¹⁰

Affiliations

¹ Department of Chemical and Biomolecular Engineering, Johns Hopkins University, Baltimore, MD 21218, USA.
² Department of Biomedical Engineering, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA; Department of Biology, School of Science, King Mongkut's Institute of Technology Ladkrabang, Bangkok 10520, Thailand.
³ Department of Biomedical Engineering, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.
⁴ Department of Chemical and Biomolecular Engineering, Johns Hopkins University, Baltimore, MD 21218, USA; Department of Radiology and Radiological Science, Johns Hopkins University School of Medicine, Baltimore, MD 21287, USA.
⁵ Department of Molecular Microbiology and Immunology, Johns Hopkins University School of Public Health, Baltimore, MD 21205, USA.
⁶ Department of Physics and Astronomy, University of New Mexico, Albuquerque, NM 87131, USA.
⁷ Department of Medicine, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.
⁸ Department of Microbiology and Immunology, Institute of Marine and Environmental Technology (IMET), University of Maryland School of Medicine, Baltimore, MD 21201, USA.
⁹ Department of Radiology and Radiological Science, Johns Hopkins University School of Medicine, Baltimore, MD 21287, USA; Bloomberg∼Kimmel Institute for Cancer Immunotherapy, Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University School of Medicine, Baltimore, MD 21231, USA.
¹⁰ Department of Chemical and Biomolecular Engineering, Johns Hopkins University, Baltimore, MD 21218, USA; Department of Biomedical Engineering, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA; Bloomberg∼Kimmel Institute for Cancer Immunotherapy, Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University School of Medicine, Baltimore, MD 21231, USA; Translational Tissue Engineering Center, Johns Hopkins University School of Medicine, Baltimore, MD 21231, USA; Department of Oncology, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA; Department of Ophthalmology, Wilmer Eye Institute, Johns Hopkins University School of Medicine, Baltimore, MD 21231, USA. Electronic address: jamie.spangler@jhu.edu.

PMID: 38547863
PMCID: PMC11102303 (available on 2025-05-16)
DOI: 10.1016/j.chembiol.2024.02.014

Abstract

Programmed death-ligand 1 (PD-L1) drives inhibition of antigen-specific T cell responses through engagement of its receptor programmed death-1 (PD-1) on activated T cells. Overexpression of these immune checkpoint proteins in the tumor microenvironment has motivated the design of targeted antibodies that disrupt this interaction. Despite clinical success of these antibodies, response rates remain low, necessitating novel approaches to enhance performance. Here, we report the development of antibody fusion proteins that block immune checkpoint pathways through a distinct mechanism targeting molecular trafficking. By engaging multiple receptor epitopes on PD-L1, our engineered multiparatopic antibodies induce rapid clustering, internalization, and degradation in an epitope- and topology-dependent manner. The complementary mechanisms of ligand blockade and receptor downregulation led to more durable immune cell activation and dramatically reduced PD-L1 availability in mouse tumors. Collectively, these multiparatopic antibodies offer mechanistic insight into immune checkpoint protein trafficking and how it may be manipulated to reprogram immune outcomes.

Keywords: PD-L1; antibody; cancer; downregulation; immune checkpoint blockade; immunotherapy; molecular trafficking; multispecific.

MeSH terms

Animals
B7-H1 Antigen* / antagonists & inhibitors
B7-H1 Antigen* / immunology
B7-H1 Antigen* / metabolism
Cell Line, Tumor
Down-Regulation* / drug effects
Female
Humans
Mice
Mice, Inbred C57BL
Tumor Microenvironment / drug effects
Tumor Microenvironment / immunology

Substances

CD274 protein, human

Abstract

MeSH terms

Substances

Grants and funding