Targeted protein degradation via chimeric antigen receptor (CAR)-mediated antigen endocytosis in T cells.
7 affiliations
- 1State Key Laboratory of Molecular Oncology, Tsinghua University, Beijing, China.
- 2Institute for Immunology, Tsinghua University, Beijing, China.
- 3School of Basic Medical Sciences, Tsinghua Medicine, Tsinghua University, Beijing, China.
- 4Beijing Key Laboratory of Immunological Research of Allergy, Tsinghua University, Beijing, China.
- 5Tsinghua-Peking Center for Life Sciences, Beijing, China.
- 6Peking University-Tsinghua University-National Institute of Biological Sciences (PTN) Joint Graduate Program, Academy for Advanced Interdisciplinary Studies, Peking University, Beijing, China.
- 7SXMU-Tsinghua Collaborative Innovation Center for Frontier Medicine, Shanxi Medical University, Shanxi, China.
Abstract
Targeted protein degradation (TPD) holds significant therapeutic potential over conventional biologics. However, existing platforms are constrained by their reliance on host protein degradation machinery and limited durability, rendering them inadequate for chronic diseases requiring sustained treatment. Here, we constructed a chimeric antigen receptor (CAR) incorporating the TNFR1 ectodomain as the antigen-binding domain (TNFR1T cells) and used CRISPR-mediated knockout of BCOR and ZC3H12A to generate persistent TNFR1TIF cells. TNF binding, endocytosis, and degradation were assessed in vitro by flow cytometry and immunofluorescence. Immunocompetent syngeneic, Tnf -/-, and hTNF-tg rheumatoid arthritis (RA) mice (n = 4-9) were used in vivo to evaluate engraftment, persistence, and disease severity (clinical scoring and grip strength), with adalimumab (Humira, 1 or 10 mg/kg) as a comparator. Safety was assessed by intravenous Listeria monocytogenes challenge (1 × 10⁴ colony-forming units [CFU], n = 6) and Thy1.1-based depletion (0.25 mg, n = 3). We found that TNFR1T cells specifically bound, endocytosed, and degraded soluble TNF in vitro. Without lymphodepletion preconditioning, TNFR1TIF cells expanded and persisted for one year in immunocompetent mice. A single infusion into hTNF-tg mice reduced serum hTNF to near wild-type levels, preventing and treating all stages of RA with superior efficacy and durability than the repeated high-dose adalimumab. Antibacterial defense remained uncompromised, and anti-Thy1.1 antibody efficiently eliminated TNFR1TIF cells in vivo. This approach extends CAR-T cell targeting from cellular antigens to soluble extracellular proteins, establishing a host-machinery-independent, durable cellular-TPD platform for chronic inflammatory diseases.
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BibTeX
@article{wang2026targeted,
title = {{Targeted protein degradation via chimeric antigen receptor (CAR)-mediated antigen endocytosis in T cells.}},
author = {Youguang Wang and Zihao He and Na Yin and Min Peng},
journal = {HLife},
year = {2026},
month = may,
doi = {10.1016/j.hlife.2026.05.003},
url = {https://europepmc.org/article/MED/42903904},
note = {CC BY-NC-ND},
}
RIS
TY - JOUR
TI - Targeted protein degradation via chimeric antigen receptor (CAR)-mediated antigen endocytosis in T cells.
AU - Youguang Wang
AU - Zihao He
AU - Na Yin
AU - Min Peng
PY - 2026
DA - 2026/05/20
JO - HLife
DO - 10.1016/j.hlife.2026.05.003
UR - https://europepmc.org/article/MED/42903904
AB - Targeted protein degradation (TPD) holds significant therapeutic potential over conventional biologics. However, existing platforms are constrained by their reliance on host protein degradation machinery and limited durability, rendering them inadequate for chronic diseases requiring sustained treatment. Here, we constructed a chimeric antigen receptor (CAR) incorporating the TNFR1 ectodomain as the antigen-binding domain (TNFR1T cells) and used CRISPR-mediated knockout of BCOR and ZC3H12A to generate persistent TNFR1TIF cells. TNF binding, endocytosis, and degradation were assessed in vitro by flow cytometry and immunofluorescence. Immunocompetent syngeneic, Tnf -/-, and hTNF-tg rheumatoid arthritis (RA) mice (n = 4-9) were used in vivo to evaluate engraftment, persistence, and disease severity (clinical scoring and grip strength), with adalimumab (Humira, 1 or 10 mg/kg) as a comparator. Safety was assessed by intravenous Listeria monocytogenes challenge (1 × 10⁴ colony-forming units [CFU], n = 6) and Thy1.1-based depletion (0.25 mg, n = 3). We found that TNFR1T cells specifically bound, endocytosed, and degraded soluble TNF in vitro. Without lymphodepletion preconditioning, TNFR1TIF cells expanded and persisted for one year in immunocompetent mice. A single infusion into hTNF-tg mice reduced serum hTNF to near wild-type levels, preventing and treating all stages of RA with superior efficacy and durability than the repeated high-dose adalimumab. Antibacterial defense remained uncompromised, and anti-Thy1.1 antibody efficiently eliminated TNFR1TIF cells in vivo. This approach extends CAR-T cell targeting from cellular antigens to soluble extracellular proteins, establishing a host-machinery-independent, durable cellular-TPD platform for chronic inflammatory diseases.
SN - 2949-9283
ER -