Targeting triple-negative breast cancer using cord-blood CD34⁺ HSPC-derived mesothelin-specific CAR-NKT cells with potent antitumor activity

Yan-Ruide Li; Xinyuan Shen; Yichen Zhu; Zhe Li; Ryan Hon; Yanxin Tian; Jie Huang; Annabel S Zhao; Nathan Y Ma; Catherine Zhang; David Lin; Karine Sargsyan; Yuan Yuan; Lili Yang

doi:10.1186/s13045-025-01736-9

Targeting triple-negative breast cancer using cord-blood CD34⁺ HSPC-derived mesothelin-specific CAR-NKT cells with potent antitumor activity

J Hematol Oncol. 2025 Oct 13;18(1):86. doi: 10.1186/s13045-025-01736-9.

Authors

Yan-Ruide Li^{1

2}, Xinyuan Shen^{1

2}, Yichen Zhu^{1

2}, Zhe Li^{1

2}, Ryan Hon^{1

2}, Yanxin Tian^{1

2}, Jie Huang^{1

2}, Annabel S Zhao^{1

2}, Nathan Y Ma^{1

2}, Catherine Zhang^{1

2}, David Lin³, Karine Sargsyan⁴, Yuan Yuan³, Lili Yang^{5

6

7

8

9

10

11}

Affiliations

¹ Department of Microbiology, Immunology & Molecular Genetics, University of California, Los Angeles, Los Angeles, CA, 90095, USA.
² Department of Bioengineering, University of California, Los Angeles, Los Angeles, CA, 90095, USA.
³ Division of Medical Oncology, Cedars-Sinai Medical Center, Los Angeles, CA, 90048, USA.
⁴ OncoBiobank, Cedars-Sinai Medical Center, Los Angeles, CA, 90048, USA.
⁵ Department of Microbiology, Immunology & Molecular Genetics, University of California, Los Angeles, Los Angeles, CA, 90095, USA. liliyang@ucla.edu.
⁶ Department of Bioengineering, University of California, Los Angeles, Los Angeles, CA, 90095, USA. liliyang@ucla.edu.
⁷ Eli and Edythe Broad Center of Regenerative Medicine and Stem Cell Research, University of California, Los Angeles, Los Angeles, CA, 90095, USA. liliyang@ucla.edu.
⁸ Jonsson Comprehensive Cancer Center, David Geffen School of Medicine, University of California, Los Angeles, Los Angeles, CA, 90095, USA. liliyang@ucla.edu.
⁹ Molecular Biology Institute, University of California, Los Angeles, Los Angeles, CA, 90095, USA. liliyang@ucla.edu.
¹⁰ Parker Institute for Cancer Immunotherapy, University of California, Los Angeles, Los Angeles, CA, 90095, USA. liliyang@ucla.edu.
¹¹ Goodman-Luskin Microbiome Center, University of California, Los Angeles, Los Angeles, CA, 90095, USA. liliyang@ucla.edu.

Abstract

Background: Triple-negative breast cancer (TNBC) is an aggressive subtype of breast cancer characterized by the lack of ER, PR, and HER2 expression. Its aggressive behavior, high degree of tumor heterogeneity, and immunosuppressive tumor microenvironment (TME) are associated with poor clinical outcomes, rapid disease progression, and limited therapeutic options. Although chimeric antigen receptor (CAR)-engineered T cell therapy has shown certain promise, its applicability in TNBC is hindered by antigen escape, TME-mediated suppression, and the logistical constraints of autologous cell production.

Methods: In this study, we employed hematopoietic stem and progenitor cell (HSPC) gene engineering and a feeder-free HSPC differentiation culture to generate allogeneic IL-15-enhanced, mesothelin-specific CAR-engineered invariant natural killer T (^Allo15MCAR-NKT) cells.

Results: These cells demonstrated robust and multifaceted antitumor activity against TNBC, mediated by CAR- and NK receptor-dependent cytotoxicity, as well as selective targeting of CD1d⁺ TME immunosuppressive cells through their TCR. In both orthotopic and metastatic TNBC xenograft models, ^Allo15MCAR-NKT cells demonstrated potent antitumor activity, associated with robust effector and cytotoxic phenotypes, low exhaustion, and a favorable safety profile without inducing graft-versus-host disease.

Conclusions: Together, these results support ^Allo15MCAR-NKT cells as a next-generation, off-the-shelf immunotherapy with strong therapeutic potential for TNBC, particularly in the context of metastasis, immune evasion, and treatment resistance.

Keywords: Allogeneic CAR-NKT cells; Allogeneic cell therapy; Allorejection; CRISPR-Cas9 gene editing; HLA ablation; Mesothelin-targeting CAR (MCAR); Metastatic model; Multiple tumor targeting mechanism; Off-the-shelf; Orthotopic model; Potent antitumor activity; Triple-negative breast cancer (TNBC); Tumor microenvironment (TME); Tumor-associated macrophage (TAM).

Publication types

Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

Animals
Antigens, CD34 / immunology
Antigens, CD34 / metabolism
Cell Line, Tumor
Female
Fetal Blood / cytology
GPI-Linked Proteins* / immunology
Hematopoietic Stem Cells* / cytology
Hematopoietic Stem Cells* / immunology
Humans
Immunotherapy, Adoptive* / methods
Mesothelin
Mice
Mice, Inbred NOD
Mice, SCID
Natural Killer T-Cells* / immunology
Natural Killer T-Cells* / transplantation
Receptors, Chimeric Antigen* / immunology
Triple Negative Breast Neoplasms* / immunology
Triple Negative Breast Neoplasms* / pathology
Triple Negative Breast Neoplasms* / therapy
Xenograft Model Antitumor Assays

Substances

Mesothelin
GPI-Linked Proteins
Receptors, Chimeric Antigen
Antigens, CD34

Abstract

Publication types

MeSH terms

Substances

Grants and funding