A tumor-on-a-chip for in vitro study of CAR-T cell immunotherapy in solid tumors

Haijiao Liu^{1

2}, Estela Noguera-Ortega³, Xuanqi Dong¹, Won Dong Lee^{4

5

6}, Jeehan Chang^{1

2}, Sezin Aday Aydin¹, Yumei Li⁷, Yonghee Shin^{8

9}, Xinyi Shi³, Maria Liousia³, Marina C Martinez³, Joshua J Brotman³, Soyeon Kim³, Zeyu Chen^{10

11}, Anni Wang¹, Zirui Ou¹, Jungwook Paek¹, Ju Young Park¹, Aidi Liu¹, Haonan Hu¹, Zebin Xiao¹², Dora Maria Racca¹, Se-Jeong Kim¹, G Scott Worthen¹³, Wei Guo⁷, Ellen Puré^{10

12

14}, Taewook Kang⁸, Joshua D Rabinowitz^{4

5}, E John Wherry^{10

11}, Edmund K Moon³, Steven M Albelda³, Dan Dongeun Huh^{15

16

17}

Affiliations

¹ Department of Bioengineering, University of Pennsylvania, Philadelphia, PA, USA.
² NSF Science and Technology Center for Engineering Mechanobiology, University of Pennsylvania, Philadelphia, PA, USA.
³ Pulmonary, Allergy and Critical Care Division, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA.
⁴ Lewis Sigler Institute for Integrative Genomics, Princeton University, Princeton, NJ, USA.
⁵ Department of Chemistry, Princeton University, Princeton, NJ, USA.
⁶ Department of Biochemistry, Yonsei University, Seoul, Republic of Korea.
⁷ Department of Biology, University of Pennsylvania, Philadelphia, PA, USA.
⁸ Department of Chemical and Biomolecular Engineering, Sogang University, Seoul, Republic of Korea.
⁹ Department of Biologics, Gachon University, Incheon, Republic of Korea.
¹⁰ Department of Systems Pharmacology and Translational Therapeutics, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA.
¹¹ Institute for Immunology, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA.
¹² Department of Biomedical Sciences, School of Veterinary Medicine, University of Pennsylvania, Philadelphia, PA, USA.
¹³ Division of Hematology, Department of Pediatrics, Children's Hospital of Philadelphia, Philadelphia, PA, USA.
¹⁴ Institute for Regenerative Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA.
¹⁵ Department of Bioengineering, University of Pennsylvania, Philadelphia, PA, USA. huhd@seas.upenn.edu.
¹⁶ NSF Science and Technology Center for Engineering Mechanobiology, University of Pennsylvania, Philadelphia, PA, USA. huhd@seas.upenn.edu.
¹⁷ Institute for Regenerative Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA. huhd@seas.upenn.edu.

PMID: 41107533
DOI: 10.1038/s41587-025-02845-z

A tumor-on-a-chip for in vitro study of CAR-T cell immunotherapy in solid tumors

Haijiao Liu et al. Nat Biotechnol. 2025.

. 2025 Oct 17.

doi: 10.1038/s41587-025-02845-z. Online ahead of print.

Authors

Affiliations

¹ Department of Bioengineering, University of Pennsylvania, Philadelphia, PA, USA.
² NSF Science and Technology Center for Engineering Mechanobiology, University of Pennsylvania, Philadelphia, PA, USA.
³ Pulmonary, Allergy and Critical Care Division, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA.
⁴ Lewis Sigler Institute for Integrative Genomics, Princeton University, Princeton, NJ, USA.
⁵ Department of Chemistry, Princeton University, Princeton, NJ, USA.
⁶ Department of Biochemistry, Yonsei University, Seoul, Republic of Korea.
⁷ Department of Biology, University of Pennsylvania, Philadelphia, PA, USA.
⁸ Department of Chemical and Biomolecular Engineering, Sogang University, Seoul, Republic of Korea.
⁹ Department of Biologics, Gachon University, Incheon, Republic of Korea.
¹⁰ Department of Systems Pharmacology and Translational Therapeutics, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA.
¹¹ Institute for Immunology, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA.
¹² Department of Biomedical Sciences, School of Veterinary Medicine, University of Pennsylvania, Philadelphia, PA, USA.
¹³ Division of Hematology, Department of Pediatrics, Children's Hospital of Philadelphia, Philadelphia, PA, USA.
¹⁴ Institute for Regenerative Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA.
¹⁵ Department of Bioengineering, University of Pennsylvania, Philadelphia, PA, USA. huhd@seas.upenn.edu.
¹⁶ NSF Science and Technology Center for Engineering Mechanobiology, University of Pennsylvania, Philadelphia, PA, USA. huhd@seas.upenn.edu.
¹⁷ Institute for Regenerative Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA. huhd@seas.upenn.edu.

PMID: 41107533
DOI: 10.1038/s41587-025-02845-z

Abstract

Our limited understanding of cancer-immune interactions remains a critical barrier to advancing chimeric antigen receptor (CAR)-T cell therapy for solid malignancies. Here, we present a microengineered system that enables vascularization of human tumor explants and their controlled perfusion with immune cells to model the activity of CAR-T cells in the tumor microenvironment. Using vascularized human lung adenocarcinoma tumors, we first demonstrate the ability of our tumor-on-a-chip system to simulate, visualize and interrogate CAR-T cell function. We then test a chemokine-directed CAR-T cell engineering strategy in a model of malignant pleural mesothelioma and validate our findings in a matching in vivo mouse model. Finally, we describe a potential therapeutic target that can be pharmacologically modulated to increase the efficacy of CAR-T cells in lung adenocarcinoma, for which we present biomarkers identified by global metabolomics analysis. Our microphysiological system provides promising in vitro technology to advance the development of adoptive cell therapies for cancer and other diseases.

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Conflict of interest statement

Competing interests: D.D.H. is a co-founder of Vivodyne and holds equity in the company. D.D.H. and H.L. are inventors on a patent application for tumor-on-a-chip technology. E.J.W. holds equity and has other ownership interests in Arseal Bio, Danger Bio and Surface Oncology. E.J.W. has consulting or advisory role at Danger Bio, Jaenssen, Marengo Therapeutics, NewLimit, Pluto Immunotherapeutics, Related Sciences, Santa Ana Bio, Surface Oncology and Synthekine. S.M.A. is a scientific founder and holds equity in Capstan Therapeutics. S.M.A. is on the scientific advisory boards of Verismo and Bio4t2. E.P. is a scientific founder and holds equity in Capstan Therapeutics. E.P. is on the scientific advisory boards of Parthenon Therapeutics and POINT Biopharma. E.P. is an inventor (University of Pennsylvania) on a patent (10329355) and patent application for the 4G5 FAP CAR (Patent Applications 20210087294 and 20210087295). E.P. is an inventor (University of Pennsylvania) on a patent for the use of CAR-T therapy in heart disease (US Provisional Patent Application 62/563,323 filed 26 September 2017, WIPO Patent Application PCT/US2018/052605). J.D.R. is an advisor and stockholder in Colorado Research Partners, L.E.A.F. Pharmaceuticals, Bantam Pharmaceuticals, Barer Institute and Rafael Pharmaceuticals; a paid consultant of Pfizer and Third Rock Ventures; a founder, director, and stockholder of Farber Partners, Serien Therapeutics and Sofro Pharmaceuticals; a founder and stockholder in Empress Therapeutics; inventor of patents held by Princeton University; and a director of the Princeton University-PKU Shenzhen collaboration. The remaining authors declare no competing interests.

References

1. Newick, K., O’Brien, S., Moon, E. & Albelda, S. M. CAR T cell therapy for solid tumors. Annu Rev. Med. 68, 139–152 (2017). - PubMed - DOI
1. Shah, N. N. & Fry, T. J. Mechanisms of resistance to CAR T cell therapy. Nat. Rev. Clin. Oncol. 16, 372–385 (2019). - PubMed - PMC
1. Lim, W. A. & June, C. H. The principles of engineering immune cells to treat cancer. Cell 168, 724–740 (2017). - PubMed - PMC - DOI
1. Joyce, J. A. & Fearon, D. T. T cell exclusion, immune privilege, and the tumor microenvironment. Science (1979) 348, 74–80 (2015).
1. Ramakrishna, S., Barsan, V. & Mackall, C. Prospects and challenges for use of CAR T cell therapies in solid tumors. Expert Opin. Biol. Ther. 20, 503–516 (2020). - PubMed - DOI

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A tumor-on-a-chip for in vitro study of CAR-T cell immunotherapy in solid tumors

Affiliations

A tumor-on-a-chip for in vitro study of CAR-T cell immunotherapy in solid tumors

Authors

Affiliations

Abstract

Conflict of interest statement

References

Grants and funding

LinkOut - more resources

Full Text Sources