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CSPG4-Specific CAR.CIK Lymphocytes as a Novel Therapy for the Treatment of Multiple Soft-Tissue Sarcoma Histotypes
Valeria Leuci1,2, Chiara Donini1,2, Giovanni Grignani1
1Candiolo Cancer Institute, FPO-IRCCS, Candiolo, Turin, Italy.
Purpose:
No effective therapy is available for unresectable soft-tissue sarcomas (STS). This unmet clinical need prompted us to test whether chondroitin sulfate proteoglycan 4 (CSPG4)-specific chimeric antigen receptor (CAR)-redirected cytokine-induced killer lymphocytes (CAR.CIK) are effective in eliminating tumor cells derived from multiple STS histotypes in vitro and in immunodeficient mice.
Experimental Design:
The experimental platform included patient-derived CAR.CIK and cell lines established from multiple STS histotypes. CAR.CIK were transduced with a retroviral vector encoding second-generation CSPG4-specific CAR (CSPG4-CAR) with 4-1BB costimulation. The functional activity of CSPG4-CAR.CIK was explored in vitro, in two- and three-dimensional STS cultures, and in three in vivo STS xenograft models.
Results:
CSPG4-CAR.CIK were efficiently generated from patients with STS. CSPG4 was highly expressed in multiple STS histotypes by in silico analysis and on all 16 STS cell lines tested by flow cytometry. CSPG4-CAR.CIK displayed superior in vitro cytolytic activity against multiple STS histotypes as compared with paired unmodified control CIK. CSPG4-CAR.CIK also showed strong antitumor activity against STS spheroids; this effect was associated with tumor recruitment, infiltration, and matrix penetration. CSPG4-CAR.CIK significantly delayed or reversed tumor growth in vivo in three STS xenograft models (leiomyosarcoma, undifferentiated pleomorphic sarcoma, and fibrosarcoma). Tumor growth inhibition persisted for up to 2 weeks following the last administration of CSPG4-CAR.CIK.
Conclusions:
This study has shown that CSPG4-CAR.CIK effectively targets multiple STS histotypes in vitro and in immunodeficient mice. These results provide a strong rationale to translate the novel strategy we have developed into a clinical setting.
Insights
Chimeric antigen receptor-redirected cytokine-induced killer lymphocytes (CAR.CIK) targeting chondroitin sulfate proteoglycan 4 (CSPG4) show promise for treating soft-tissue sarcomas (STS). This novel therapy effectively eliminated STS tumor cells in vitro and in vivo models.
Area of Science:
- Oncology
- Immunotherapy
- Cancer Biology
Background:
- Unresectable soft-tissue sarcomas (STS) lack effective therapeutic options, representing a significant unmet clinical need.
- Chondroitin sulfate proteoglycan 4 (CSPG4) is a cell surface proteoglycan highly expressed across multiple STS histotypes.
Purpose of the Study:
- To evaluate the efficacy of chondroitin sulfate proteoglycan 4 (CSPG4)-specific chimeric antigen receptor (CAR)-redirected cytokine-induced killer lymphocytes (CAR.CIK) against various STS histotypes.
- To assess the in vitro and in vivo anti-tumor activity of CSPG4-CAR.CIK in preclinical models.
Main Methods:
- Generation of patient-derived CSPG4-CAR.CIK using retroviral transduction with a second-generation CSPG4-specific CAR construct incorporating 4-1BB costimulation.
- In vitro assessment of CAR.CIK cytolytic activity against STS cell lines and spheroids in 2D and 3D cultures.
- In vivo evaluation of CSPG4-CAR.CIK efficacy in three distinct STS xenograft models in immunodeficient mice.
Main Results:
- CSPG4-CAR.CIK were efficiently generated and demonstrated superior in vitro cytolytic activity against multiple STS histotypes compared to unmodified CIK cells.
- CSPG4-CAR.CIK exhibited significant antitumor effects against STS spheroids, including tumor recruitment, infiltration, and matrix penetration.
- In vivo studies showed that CSPG4-CAR.CIK significantly delayed or reversed tumor growth in leiomyosarcoma, undifferentiated pleomorphic sarcoma, and fibrosarcoma xenograft models, with effects persisting for up to two weeks post-treatment.
Conclusions:
- CSPG4-CAR.CIK therapy effectively targets and eliminates diverse STS histotypes in vitro and in vivo.
- These findings provide a strong preclinical rationale for the clinical translation of CSPG4-CAR.CIK as a novel therapeutic strategy for unresectable STS.
