A novel strategy of co-expressing CXCR5 and IL-7 enhances CAR-T cell effectiveness in osteosarcoma

Xinhui Hui; Muhammad Asad Farooq; Yiran Chen; Iqra Ajmal; Yaojun Ren; Min Xue; Yuzhou Ji; Bingtan Du; Shijia Wu; Wenzheng Jiang

doi:10.3389/fimmu.2024.1462076

A novel strategy of co-expressing CXCR5 and IL-7 enhances CAR-T cell effectiveness in osteosarcoma

Front Immunol. 2024 Oct 10:15:1462076. doi: 10.3389/fimmu.2024.1462076. eCollection 2024.

Authors

Xinhui Hui^#¹, Muhammad Asad Farooq^#^{1

2}, Yiran Chen¹, Iqra Ajmal^{1

2}, Yaojun Ren^{1

3}, Min Xue¹, Yuzhou Ji¹, Bingtan Du¹, Shijia Wu¹, Wenzheng Jiang¹

Affiliations

¹ Shanghai Key Laboratory of Regulatory Biology, School of Life Sciences, East China Normal University, Shanghai, China.
² The First Dongguan Affiliated Hospital, Guangdong Provincial Key Laboratory of Medical Immunology and Molecular Diagnostics, Guangdong Medical University, Dongguan, China.
³ College of Life Science, Xinjiang Normal University, Urumqi, China.

^# Contributed equally.

Abstract

Background: Solid tumors are characterized by a low blood supply, complex stromal architecture, and immunosuppressive milieu, which inhibit CAR-T cell entry and survival. CXCR5 has previously been employed to increase CAR-T cell infiltration into CXCL13+ cancers. On the other hand, IL-7 improves the survival and persistence of T cells inside a solid tumor milieu.

Methods: We constructed a novel NKG2D-based CAR (C5/IL7-CAR) that co-expressed CXCR5 and IL-7. The human osteosarcoma cell lines U-2 OS, 143B, and Mg63 highly expressed MICA/B and CXCL13, thus presenting a perfect avenue for the present study.

Results: Novel CAR-T cells are superior in their activation, degranulation, and cytokine release competence, hence lysing more target cells than conventional CAR. Furthermore, CXCR5 and IL-7 co-expression decreased the expression of PD-1, TIM-3, and TIGIT and increased Bcl-2 expression. Novel CAR-T cells show enhanced proliferation and differentiation towards the stem cell memory T cell phenotype. C5/IL7-CAR-T cells outperformed conventional CAR-T in eradicating osteosarcoma in mouse models and displayed better survival. Additionally, CXCR5 and IL-7 co-expression enhanced CAR-T cell numbers, cytokine release, and survival in implanted tumor tissues compared to conventional CAR-T cells. Mechanistically, C5/IL7-CAR-T cells displayed enhanced STAT5 signaling.

Conclusion: These findings highlight the potential of CXCR5 and IL-7 co-expression to improve CAR-T cell therapy efficacy against osteosarcoma.

Keywords: CAR-T therapy; CXCR5; IL-7; T cell migration; osteosarcoma; tumor microenvironment.

MeSH terms

Animals
Bone Neoplasms* / immunology
Bone Neoplasms* / pathology
Bone Neoplasms* / therapy
Cell Line, Tumor
Humans
Immunotherapy, Adoptive* / methods
Interleukin-7* / genetics
Interleukin-7* / immunology
Interleukin-7* / metabolism
Mice
Osteosarcoma* / immunology
Osteosarcoma* / pathology
Osteosarcoma* / therapy
Receptors, CXCR5* / metabolism
Receptors, Chimeric Antigen* / genetics
Receptors, Chimeric Antigen* / immunology
Receptors, Chimeric Antigen* / metabolism
T-Lymphocytes / immunology
T-Lymphocytes / metabolism
Xenograft Model Antitumor Assays

Substances

Interleukin-7
Receptors, CXCR5
CXCR5 protein, human
Receptors, Chimeric Antigen
IL7 protein, human

Grants and funding

The author(s) declare financial support was received for the research, authorship, and/or publication of this article. This work was supported by the Science and Technology Commission of Shanghai Municipality (21S11906200, 201409002900), the National Natural Science Foundation of China (81771306), the National Key Research and Development Program of China (2021YFF0702401), NMPA Key Laboratory for Quality Control of Therapeutic Monoclonal Antibodies (2023-DK-01).