Engineering an in vivo charging station for CAR-redirected invariant natural killer T cells to enhance cancer therapy

Posted on
engineering-an-in-vivo-charging-station-for-car-redirected-invariant-natural-killer-t-cells-to-enhance-cancer-therapy
Engineering an in vivo charging station for CAR-redirected invariant natural killer T cells to enhance cancer therapy

Data availability

All data are available within the Article, Supplementary Information or Source Data file. The corresponding authors (L.Y. and S.L.) will make raw data and step-by-step protocols available upon request. All data generated in this study, including source data for figures, are available via figshare at (ref. 91). Source data are provided with this paper.

Code availability

The scRNA-seq data generated in this study have been deposited in the public repository Gene Expression Omnibus Database under accession GSE291443.

References

  1. Bendelac, A., Savage, P. B. & Teyton, L. The biology of NKT cells. Annu. Rev. Immunol. 25, 297–336 (2007).

    Article  CAS  PubMed  Google Scholar 

  2. Kronenberg, M. Toward an understanding of NKT cell biology: progress and paradoxes. Annu. Rev. Immunol. 23, 877–900 (2005).

    Article  CAS  PubMed  Google Scholar 

  3. Bae, E. A., Seo, H., Kim, I. K., Jeon, I. & Kang, C. Y. Roles of NKT cells in cancer immunotherapy. Arch. Pharm. Res. 42, 543–548 (2019).

    Article  CAS  PubMed  Google Scholar 

  4. Li, Y.-R., Zhou, K., Zhu, Y., Halladay, T. & Yang, L. Breaking the mold: unconventional T cells in cancer therapy. Cancer Cell 43, 317–322 (2025).

    Article  CAS  PubMed  Google Scholar 

  5. Shissler, S. C. & Webb, T. J. The ins and outs of type I iNKT cell development. Mol. Immunol. 105, 116–130 (2019).

    Article  CAS  PubMed  Google Scholar 

  6. Tachibana, T. et al. Increased intratumor Valpha24-positive natural killer T cells: a prognostic factor for primary colorectal carcinomas. Clin. Cancer Res. 11, 7322–7327 (2005).

    Article  CAS  PubMed  Google Scholar 

  7. Molling, J. W. et al. Low levels of circulating invariant natural killer T cells predict poor clinical outcome in patients with head and neck squamous cell carcinoma. J. Clin. Oncol. 25, 862–868 (2007).

    Article  PubMed  Google Scholar 

  8. Muhammad Ali Tahir, S. et al. Loss of IFN-γ production by invariant NK T cells in advanced cancer. J. Immunol. 167, 4046–4050 (2001).

    Article  Google Scholar 

  9. Dhodapkar, M. V. et al. A reversible defect in natural killer T cell function characterizes the progression of premalignant to malignant multiple myeloma. J. Exp. Med. 197, 1667–1676 (2003).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Zhou, X. et al. CAR-redirected natural killer T cells demonstrate superior antitumor activity to CAR-T cells through multimodal CD1d-dependent mechanisms. Nat. Cancer. 5, 1607–1607 (2024).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Li, Y.-R. et al. Generation of allogeneic CAR-NKT cells from hematopoietic stem and progenitor cells using a clinically guided culture method. Nat. Biotechnol. 43, 329–344 (2025).

    Article  CAS  PubMed  Google Scholar 

  12. Li, Y.-R. et al. Generating allogeneic CAR-NKT cells for off-the-shelf cancer immunotherapy with genetically engineered HSP cells and feeder-free differentiation culture. Nat. Protoc. 20, 1352–1388 (2025).

    Article  CAS  PubMed  Google Scholar 

  13. Li, Y.-R. et al. Allogeneic CD33-directed CAR-NKT cells for the treatment of bone marrow-resident myeloid malignancies. Nat. Commun. 16, 1248 (2025).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Rotolo, A. et al. Enhanced anti-lymphoma activity of CAR19-iNKT cells underpinned by dual CD19 and CD1d targeting. Cancer Cell 34, 596–610.e11 (2018).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Ramos, C. A. et al. Off-the-shelf CD19-specific CAR-NKT cells in patients with relapsed or refractory B-cell malignancies. Transplant. Cell. Ther. 30, S41–S42 (2024).

    Article  Google Scholar 

  16. Li, Y.-R. et al. Development of allogeneic HSC-engineered iNKT cells for off-the-shelf cancer immunotherapy. Cell Rep. Med. 2, 100449 (2021).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Li, Y.-R. et al. Engineering allorejection-resistant CAR-NKT cells from hematopoietic stem cells for off-the-shelf cancer immunotherapy. Mol. Ther. 32, 1849–1874 (2024).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Ren, H. et al. Bispecific CAR-iNKT immunotherapy for high risk MLL-rearranged acute lymphoblastic leukemia. Blood 142, 766 (2023).

    Article  Google Scholar 

  19. Xu, X. et al. NKT cells coexpressing a GD2-specific chimeric antigen receptor and IL15 show enhanced in vivo persistence and antitumor activity against neuroblastoma. Clin. Cancer Res. 25, 7126–7138 (2019).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. Heczey, A. et al. Anti-GD2 CAR-NKT cells in relapsed or refractory neuroblastoma: updated phase 1 trial interim results. Nat. Med. 29, 1379–1388 (2023).

    Article  CAS  PubMed  Google Scholar 

  21. Heczey, A. et al. Anti-GD2 CAR-NKT cells in patients with relapsed or refractory neuroblastoma: an interim analysis. Nat. Med. 26, 1686–1690 (2020).

    Article  CAS  PubMed  Google Scholar 

  22. Li, Y.-R. et al. Allogeneic stem cell-engineered EGFRvIII-specific CAR-NKT cells for treating glioblastoma with enhanced efficacy and safety. Mol. Ther. 33, 6041–6062 (2025).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. Shaik, R. S. et al. Glypican-3-specific CAR-NKT cells overexpressing BATF3 mediate potent antitumor activity against hepatocellular carcinoma. J. Clin. Oncol. 40, e14521 (2025).

    Article  Google Scholar 

  24. Liu, Y. et al. IL-21-armored B7H3 CAR-iNKT cells exert potent antitumor effects. iScience 27, 108597 (2024).

    Article  CAS  PubMed  Google Scholar 

  25. Li, Y.-R. et al. Targeting orthotopic and metastatic pancreatic cancer with allogeneic stem cell-engineered mesothelin-redirected CAR-NKT cells. Proc. Natl Acad. Sci. USA 122, e2517786122 (2025).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  26. Li, Y.-R. et al. Targeting triple-negative breast cancer using cord-blood CD34+ HSPC-derived mesothelin-specific CAR-NKT cells with potent antitumor activity. J. Hematol. Oncol. 18, 86 (2025).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  27. Urakami, A. et al. 315iPSC-derived CAR-iNKT cells targeting HER2 show prolonged tumor control and promote durable survival in a tumor xenograft model. J. Immunother. Cancer https://doi.org/10.1136/jitc-2023-SITC2023.0315 (2023).

  28. Li, Y.-R. et al. Overcoming ovarian cancer resistance and evasion to CAR-T cell therapy by harnessing allogeneic CAR-NKT cells. Med 6, 100804 (2025).

    Article  CAS  PubMed  Google Scholar 

  29. Li, Y.-R. et al. Multimodal targeting of metastatic renal cell carcinoma via CD70-directed allogeneic CAR-NKT cells. Cell Rep. Med. 6, 102321 (2025).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  30. Li, Y.-R., Zhu, Y., Fang, Y., Lyu, Z. & Yang, L. Emerging trends in clinical allogeneic CAR cell therapy. Med 6, 100677 (2025).

    Article  CAS  PubMed  Google Scholar 

  31. Lyu, Z. et al. Addressing graft-versus-host disease in allogeneic cell-based immunotherapy for cancer. Exp. Hematol. Oncol. 14, 66 (2025).

    Article  PubMed  PubMed Central  Google Scholar 

  32. Pillai, A. B., George, T. I., Dutt, S., Teo, P. & Strober, S. Host NKT cells can prevent graft-versus-host disease and permit graft antitumor activity after bone marrow transplantation. J. Immunol. 178, 6242–6251 (2007).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  33. Coman, T. et al. Human CD4-invariant NKT lymphocytes regulate graft versus host disease. Oncoimmunology 7, e1470735 (2018).

    Article  PubMed  PubMed Central  Google Scholar 

  34. Li, Y.-R., Zhu, Y., Chen, Y. & Yang, L. The clinical landscape of CAR-engineered unconventional T cells. Trends Cancer 11, 520–539 (2025).

    Article  CAS  PubMed  Google Scholar 

  35. Waldowska, M., Bojarska-Junak, A. & Roliński, J. A brief review of clinical trials involving manipulation of invariant NKT cells as a promising approach in future cancer therapies. Cent. Eur. J. Immunol. 42, 181–195 (2017).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  36. Chang, D. H. et al. Sustained expansion of NKT cells and antigen-specific T cells after injection of alpha-galactosyl-ceramide loaded mature dendritic cells in cancer patients. J. Exp. Med. 201, 1503–1517 (2005).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  37. Simon, B. et al. The generation of CAR-transfected natural killer T cells for the immunotherapy of melanoma. Int. J. Mol. Sci. 19, 2365 (2018).

  38. Fujii, S.-I. et al. NKT cells as an ideal anti-tumor immunotherapeutic. Front. Immunol. 4, 409 (2013).

    Article  PubMed  PubMed Central  Google Scholar 

  39. Nelson, A., Lukacs, J. D. & Johnston, B. The current landscape of NKT cell immunotherapy and the hills ahead. Cancers 13, 5174 (2021).

  40. Li, Y.-R., Fang, Y., Lyu, Z., Zhu, Y. & Yang, L. Exploring the dynamic interplay between cancer stem cells and the tumor microenvironment: implications for novel therapeutic strategies. J. Transl. Med. 21, 686 (2023).

    Article  PubMed  PubMed Central  Google Scholar 

  41. Li, Y.-R. et al. Tumor-localized administration of α-GalCer to recruit invariant natural killer T cells and enhance their antitumor activity against solid tumors. Int. J. Mol. Sci. 23, 7547 (2022).

  42. Zhang, Y. et al. GalCer and iNKT cell-based cancer immunotherapy: realizing the therapeutic potentials. Front. Immunol. 10, 1126 (2019).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  43. Sullivan, B. A. & Kronenberg, M. Activation or anergy: NKT cells are stunned by alpha-galactosylceramide. J. Clin. Invest. 115, 2328–2329 (2005).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  44. Spada, F. M. et al. Low expression level but potent antigen presenting function of CD1d on monocyte lineage cells. Eur. J. Immunol. 30, 3468–3477 (2000).

    CAS  PubMed  Google Scholar 

  45. Liu, Z. et al. Viscoelastic synthetic antigen-presenting cells for augmenting the potency of cancer therapies. Nat. Biomed. Eng. 8, 1615–1633 (2024).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  46. Adu-Berchie, K. et al. Generation of functionally distinct T-cell populations by altering the viscoelasticity of their extracellular matrix. Nat. Biomed. Eng. 7, 1374–1391 (2023).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  47. Hasani-Sadrabadi, M. M. et al. Mechanobiological mimicry of helper T lymphocytes to evaluate cell-biomaterials crosstalk. Adv. Mater. 30, e1706780 (2018).

    Article  PubMed  Google Scholar 

  48. Sunshine, J. C., Perica, K., Schneck, J. P. & Green, J. J. Particle shape dependence of CD8+ T cell activation by artificial antigen presenting cells. Biomaterials 35, 269–277 (2014).

    Article  CAS  PubMed  Google Scholar 

  49. Chaudhuri, O. et al. Hydrogels with tunable stress relaxation regulate stem cell fate and activity. Nat. Mater. 15, 326–334 (2016).

    Article  CAS  PubMed  Google Scholar 

  50. Krijgsman, D., Hokland, M. & Kuppen, P. J. K. The role of natural killer T cells in cancer—a phenotypical and functional approach. Front. Immunol. 9, 367 (2018).

  51. Cortés-Selva, D., Dasgupta, B., Singh, S. & Grewal, I. S. Innate and innate-like cells: the future of chimeric antigen receptor (CAR) cell therapy. Trends Pharmacol. Sci. 42, 45–59 (2021).

    Article  PubMed  Google Scholar 

  52. Li, Y.-R., Zhu, Y. & Yang, L. IL-15 in CAR engineering: striking an efficacy–safety balance. Trends Mol. Med. 31, 977–981 (2025).

    Article  CAS  PubMed  Google Scholar 

  53. Gordy, L. E. et al. IL-15 regulates homeostasis and terminal maturation of NKT cells. J. Immunol. 187, 6335–6345 (2011).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  54. Lynn, R. C. et al. c-Jun overexpression in CAR T cells induces exhaustion resistance. Nature 576, 293–300 (2019).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  55. Crespo, J., Sun, H., Welling, T. H., Tian, Z. & Zou, W. T cell anergy, exhaustion, senescence, and stemness in the tumor microenvironment. Curr. Opin. Immunol. 25, 214–221 (2013).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  56. Chi, X. et al. T-cell exhaustion and stemness in antitumor immunity: characteristics, mechanisms, and implications. Front. Immunol. 14, 1104771 (2023).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  57. Liu, D. et al. IL-15 protects NKT cells from inhibition by tumor-associated macrophages and enhances antimetastatic activity. J. Clin. Invest. 122, 2221–2233 (2012).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  58. Morgan, M. A. & Schambach, A. Engineering CAR-T cells for improved function against solid tumors. Front. Immunol. 9, 2493 (2018).

    Article  PubMed  PubMed Central  Google Scholar 

  59. Li, Y.-R. et al. Advancing cell-based cancer immunotherapy through stem cell engineering. Cell Stem Cell 30, 592–610 (2023).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  60. Ma, L. et al. Enhanced CAR-T cell activity against solid tumors by vaccine boosting through the chimeric receptor. Science 365, 162–168 (2019).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  61. Anderson, A. C., Joller, N. & Kuchroo, V. K. Lag-3, Tim-3, and TIGIT: co-inhibitory receptors with specialized functions in immune regulation. Immunity 44, 989–1004 (2016).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  62. Dillekås, H., Rogers, M. S. & Straume, O. Are 90% of deaths from cancer caused by metastases? Cancer Med. 8, 5574–5576 (2019).

    Article  PubMed  PubMed Central  Google Scholar 

  63. Amini, L. et al. Preparing for CAR T cell therapy: patient selection, bridging therapies and lymphodepletion. Nat. Rev. Clin. Oncol. 19, 342–355 (2022).

    Article  PubMed  Google Scholar 

  64. Mullard, A. FDA approves first CAR T therapy. Nat. Rev. Drug Discov. 16, 669 (2017).

    PubMed  Google Scholar 

  65. Mullard, A. FDA approves second BCMA-targeted CAR-T cell therapy. Nat. Rev. Drug Discov. 21, 249 (2022).

  66. Neelapu, S. S. et al. Chimeric antigen receptor T-cell therapy – assessment and management of toxicities. Nat. Rev. Clin. Oncol. 15, 47–62 (2018).

    Article  CAS  PubMed  Google Scholar 

  67. Slezak, A. et al. Therapeutic synthetic and natural materials for immunoengineering. Chem. Soc. Rev. 53, 1789–1822 (2024).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  68. Wang, C., Ye, Y., Hu, Q., Bellotti, A. & Gu, Z. Tailoring biomaterials for cancer immunotherapy: emerging trends and future outlook. Adv. Mater. 29, 1606036 (2017).

  69. Niu, H., Zhao, P. & Sun, W. Biomaterials for chimeric antigen receptor T cell engineering. Acta Biomater. 166, 1–13 (2023).

    Article  CAS  PubMed  Google Scholar 

  70. Martin, O. A., Anderson, R. L., Narayan, K. & MacManus, M. P. Does the mobilization of circulating tumour cells during cancer therapy cause metastasis? Nat. Rev. Clin. Oncol. 14, 32–44 (2017).

    Article  CAS  PubMed  Google Scholar 

  71. Klein, C. A. Cancer progression and the invisible phase of metastatic colonization. Nat. Rev. Cancer 20, 681–694 (2020).

    Article  CAS  PubMed  Google Scholar 

  72. Steeg, P. S. Targeting metastasis. Nat. Rev. Cancer 16, 201–218 (2016).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  73. Landoni, E. et al. IL-12 reprograms CAR-expressing natural killer T cells to long-lived Th1-polarized cells with potent antitumor activity. Nat. Commun. 15, 89 (2024).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  74. Gu, X. et al. New insights into iNKT cells and their roles in liver diseases. Front. Immunol. 13, 1035950 (2022).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  75. Gustafsson, K. et al. Tumour-loaded α-type 1-polarized dendritic cells from patients with chronic lymphocytic leukaemia produce a superior NK-, NKT- and CD8+ T cell-attracting chemokine profile. Scand. J. Immunol. 74, 318–326 (2011).

    Article  CAS  PubMed  Google Scholar 

  76. Shimaoka, T. et al. Critical role for CXC chemokine ligand 16 (SR-PSOX) in Th1 response mediated by NKT cells. J. Immunol. 179, 8172–8179 (2007).

    Article  CAS  PubMed  Google Scholar 

  77. Drennan, M. B. et al. The thymic microenvironment differentially regulates development and trafficking of invariant NKT cell sublineages. J. Immunol. 193, 5960–5972 (2014).

    Article  CAS  PubMed  Google Scholar 

  78. Han, Y., Liu, D. & Li, L. PD-1/PD-L1 pathway: current researches in cancer. Am. J. Cancer Res. 10, 727–742 (2020).

    CAS  PubMed  PubMed Central  Google Scholar 

  79. Goodman, A., Patel, S. P. & Kurzrock, R. PD-1–PD-L1 immune-checkpoint blockade in B-cell lymphomas. Nat. Rev. Clin. Oncol. 14, 203–220 (2017).

    Article  CAS  PubMed  Google Scholar 

  80. Zou, W., Wolchok, J. D. & Chen, L. PD-L1 (B7-H1) and PD-1 pathway blockade for cancer therapy: mechanisms, response biomarkers, and combinations. Sci. Transl. Med. 8, 328rv4 (2016).

    Article  PubMed  PubMed Central  Google Scholar 

  81. Yue, W., Wang, T., Xie, L. & Shen, J. An injectable in situ hydrogel platform for sustained drug release against glioblastoma. J. Drug Deliv. Sci. Technol. 95, 105527 (2024).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  82. Khan, Z. M., Munson, J. M., Long, T. E., Vlaisavljevich, E. & Verbridge, S. S. Development of a synthetic, injectable hydrogel to capture residual glioblastoma and glioblastoma stem-like cells with CXCL12-mediated chemotaxis. Adv. Healthc. Mater. 12, e2300671 (2023).

    Article  PubMed  PubMed Central  Google Scholar 

  83. Ma, J. et al. Hydrogels for localized chemotherapy of liver cancer: a possible strategy for improved and safe liver cancer treatment. Drug Deliv. 29, 1457–1476 (2022).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  84. Tang, Z. et al. Intelligent hydrogel-assisted hepatocellular carcinoma therapy. Research 7, 0477 (2024).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  85. Chang, Y. et al. CAR-neutrophil mediated delivery of tumor-microenvironment responsive nanodrugs for glioblastoma chemo-immunotherapy. Nat. Commun. 14, 2266 (2023).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  86. Crosby, C. M. & Kronenberg, M. Tissue-specific functions of invariant natural killer T cells. Nat. Rev. Immunol. 18, 559–574 (2018).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  87. Smith, D. J. et al. Genetic engineering of hematopoietic stem cells to generate invariant natural killer T cells. Proc. Natl Acad. Sci. USA 112, 1523–1528 (2015).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  88. Giannoni, F. et al. Allelic exclusion and peripheral reconstitution by TCR transgenic T cells arising from transduced human hematopoietic stem/progenitor cells. Mol. Ther. 21, 1044–1054 (2013).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  89. Lyu, Z., Li, Y.-R. & Yang, L. Protocol for assessing pharmacokinetics and pharmacodynamics of human CAR-NKT cells in humanized mouse models using bioluminescence imaging. STAR Protoc. 6, 103957 (2025).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  90. Fang, Y. et al. Protocol to generate allorejection-resistant universal CAR-NKT cells and evaluate their efficacy, mechanism of action, safety, and immunogenicity. STAR Protoc. 6, 103810 (2025).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  91. Li, Y.-R. et al. Engineering an in vivo charging station for CAR-redirected invariant natural killer T cells to enhance cancer therapy. Figshare https://doi.org/10.6084/m9.figshare.30379627.v1 (2025).

Download references

Acknowledgements

We thank the University of California Los Angeles (UCLA) animal facility for providing animal support; the UCLA Translational Pathology Core Laboratory (TPCL) for providing histology support; the UCLA Technology Centre for Genomics and Bioinformatics (TCGB) facility for providing RNA-seq services; the UCLA CFAR Virology Core for providing human cells; and the UCLA BSCRC Flow Cytometry Core Facility for cell sorting support. This work was supported by a seed grant from UCLA Jonsson Comprehensive Cancer Center (to S.L. and L.Y.), an innovation award from UCLA Technology Development Group (to S.L. and L.Y.), a Partnering Opportunity for Discovery Stage Research Projects Award and a Partnering Opportunity for Translational Research Projects Award from the California Institute for Regenerative Medicine (CIRM) (DISC2-11157 and TRAN1-12250, to L.Y.), a CIRM Discovery grant (DISC2-14169 to S.L.), an NIH grant (R01GM143485 to S.L.), a Department of Defense CDMRP PRCRP Impact Award (CA200456 to L.Y.), a UCLA BSCRC Innovation Award (to L.Y.), and an Ablon Scholars Award (to L.Y.). L.Y. is a member of the UCLA Parker Institute for Cancer Immunotherapy. Y.-R.L. is a postdoctoral fellow supported by a UCLA MIMG M. John Pickett Post-Doctoral Fellow Award, a CIRM-BSCRC Postdoctoral Fellowship, a UCLA Sydney Finegold Postdoctoral Award, a UCLA Chancellor’s Award for Postdoctoral Research, and a Goodman–Luskin Microbiome Center Collaborative Research Fellowship award. E.Z. is a postdoctoral fellow supported by a T32 UCLA/Caltech Integrated Cardiometabolic Medicine for Bioengineers fellowship (NIH/NHLBI T32HL144449). We also thank the NIH Tetramer Facility for providing the tetramers.

Author information

Author notes

  1. These authors contributed equally: Yan-Ruide Li, Haochen Nan, Zeyang Liu.

Authors and Affiliations

  1. Department of Bioengineering, University of California Los Angeles, Los Angeles, CA, USA

    Yan-Ruide Li, Haochen Nan, Zeyang Liu, Ying Fang, Yichen Zhu, Zibai Lyu, Bo Zhang, Youcheng Yang, Xinyuan Shen, Yuning Chen, Lili Yang & Song Li

  2. Department of Microbiology, Immunology and Molecular Genetics, University of California Los Angeles, Los Angeles, CA, USA

    Yan-Ruide Li, Ying Fang, Yichen Zhu, Zibai Lyu, Enbo Zhu, Xinyuan Shen, Yuning Chen & Lili Yang

  3. Department of Pharmacology, University of California San Diego, La Jolla, CA, USA

    Zhengyao Shao

  4. Division of Cardiology, Department of Medicine, David Geffen School of Medicine, University of California Los Angeles, Los Angeles, CA, USA

    Enbo Zhu & Tzung Hsiai

  5. The Eli and Edythe Broad Center of Regenerative Medicine and Stem Cell Research, University of California Los Angeles, Los Angeles, CA, USA

    Lili Yang & Song Li

  6. Jonsson Comprehensive Cancer Centre, University of California Los Angeles, Los Angeles, CA, USA

    Lili Yang & Song Li

  7. Molecular Biology Institute, University of California Los Angeles, Los Angeles, CA, USA

    Lili Yang

  8. Parker Institute for Cancer Immunotherapy, University of California Los Angeles, Los Angeles, CA, USA

    Lili Yang

  9. Goodman–Luskin Microbiome Center, University of California Los Angeles, Los Angeles, CA, USA

    Lili Yang

Authors

  1. Yan-Ruide Li
  2. Haochen Nan
  3. Zeyang Liu
  4. Ying Fang
  5. Yichen Zhu
  6. Zibai Lyu
  7. Zhengyao Shao
  8. Enbo Zhu
  9. Bo Zhang
  10. Youcheng Yang
  11. Xinyuan Shen
  12. Yuning Chen
  13. Tzung Hsiai
  14. Lili Yang
  15. Song Li

Contributions

Y.-R.L., H.N. and Z. Liu designed the experiments, analysed the data and wrote the manuscript. Y.-R.L., L.Y. and S.L. conceived and oversaw the study, with suggestions from T.H., and revised the manuscript. Y.-R.L., H.N. and Z. Liu performed all experiments, with assistance from Y.F., Y.Z., Z. Lyu, Z.S., E.Z., B.Z., Y.Y., X.S. and Y.C.

Corresponding authors

Correspondence to Lili Yang or Song Li.

Ethics declarations

Competing interests

L.Y. is a scientific advisor to AlzChem and Amberstone Biosciences, and a co-founder, stockholder and advisory board member of Appia Bio. None of the declared companies contributed to or directed any of the research reported in this Article. Y.-R.L., H.N., Z. Liu, L.Y. and S.L. filed a patent application related to this work through UCLA Technology Development Group (UCLA TDG), identified as UCLA CASE NO. 2025-209-1, titled ‘Recruitment and activation of car-redirected invariant natural killer T cells’. The remaining authors declare no competing interests.

Peer review

Peer review information

Nature Biomedical Engineering thanks Xiaoping Bao, Tonya Webb and the other, anonymous, reviewer(s) for their contribution to the peer review of this work. Peer reviewer reports are available.

Additional information

Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Extended data

Extended Data Fig. 1 Mechanical characterization of iMRAS.

a, AFM indentation technique was used to characterize the mechanical properties of human APC, iMRAS, bare iMRAS, and iMRAS without nanoparticles. Representative stress relaxation curves of each sample are shown. b, Swelling of iMRAS with different components and PDMS elastomer (as a negative control) after 1, 7, and 15 days of DMEM immersion, quantified by the ratio of the volume of gels at day 1, day 7 or day 15 to their volumes before immersion (n = 6). Data are presented as the mean ± s.e.m. NS, not significant, by one-way ANOVA (b). Source data are provided as a Source Data file.

Source data

Extended Data Fig. 2 Fabrication and characterization of PLGA nanoparticles for iMRAS.

a, Schematic representation of PLGA nanoparticles fabrication steps using solvent evaporation and ultrasonic homogenization. Created in BioRender. Liu, Z. (2025) https://BioRender.com/jnuo8z1b, PLGA nanoparticles size distribution at different ultrasonic homogenization amplitude at 50%, 25%, and 10% (frequency fixed at 20 kHz) (n = 3). c, Size distribution of PLGA nanoparticles prepared with varying PLGA concentrations at 1, 5, 10, 20, and 40 mg/ml (n = 3). d, Comparison of nanoparticle size when different organic solvents dichloromethane (DCM), acetonitrile (MeCN), and tetrahydrofuran (THF) are used (n = 3). e, Encapsulation efficiency of αGC under different PLGA-to-αGC mass ratios. Based on optimal encapsulation efficiency and drug release kinetics, a ratio of 20:1 was selected for subsequent experiments (n = 4). f, Encapsulation efficiency of PLGA nanoparticles at different molecular weight (n = 4). g, Encapsulation efficiency and cumulative release profiles of αGC from PLGA nanoparticles with different lactic acid (LA) to glycolic acid (GA) ratios (n = 4). Data are presented as the mean ± s.e.m. NS, not significant; **P < 0.01, ***P < 0.001, by one-way ANOVA (d–g). In e, the center line represents the median, the box shows the 25% and 75% percentiles, and the whiskers extend to the minimum and maximum values. Source data are provided as a Source Data file.

Source data

Extended Data Fig. 3 Gene Set Enrichment Analysis (GSEA) of CAR-iNKT cell samples.

GSEA plots show the enrichment of gene signatures of proliferating, naïve, effector, cytotoxic, memory, and exhausted cells in the indicated cell clusters. Normal p value calculated as two-tailed t-test.

Extended Data Fig. 4 Longitudinal immune profiling of NKT, NK, monocyte, B and T cells following iMRAS treatment in C57BL/6 mice.

Middle-term (15 days) and long-term (45 days) effects of IMRAS loaded with or without αGC on leukocyte populations in blood (a), liver (b) and spleen (c) of C57BL/6 mice (n = 4). Data are presented as the mean ± s.e.m. NS, not significant; **P < 0.01, ***P < 0.001, ****P < 0.0001, by one-way ANOVA. Source data are provided as a Source Data file.

Source data

Extended Data Fig. 5 CAR-iNKT cell infiltration into A375-CD19 solid tumors.

a, Immunohistochemical (IHC) staining shows infiltration of human CD3⁺ CAR-iNKT or CAR-T cells into the A375-CD19 solid tumors. The tumor border and core are delineated, and the arrow indicates the direction of CAR-iNKT or CAR-T cell infiltration from the periphery toward the tumor core. The experimental design is illustrated in Fig. 5f. Tumor tissues were harvested on Day 60, at which point the tumor size was sufficient for analysis. b, Quantification of a (n = 5 from five experimental mice). Images were collected from five mice in one experiment, and the in vivo studies were independently repeated three times. Source data are provided as a Source Data file.

Source data

Extended Data Fig. 6 Superior antitumor efficacy of CAR-iNKT cells with iMRAS compared to conventional CAR-T cell in blood cancer and solid tumor xenograft models.

a-c, Comparing the in vivo antitumor capacity between CAR-iNKT + iMRAS and conventional CAR-T cells using an A375 human melanoma xenograft mouse model. a, Experimental design. b, Tumor volume measurements over time (n = 5). c, Tumor weight measurements at Day 30 (n = 5). Note that the data for the Vehicle and CAR-iNKT + iMRAS groups were also presented in the main Fig. 5e-g. d-g, Comparing the in vivo antitumor capacity between CAR-iNKT + iMRAS and conventional CAR-T cells using a Raji human lymphoma xenograft mouse model. d, Experimental design. e, BLI images showing the presence of tumor cells in CAR-T cell-treated experimental mice over time. f, Quantification of e (n = 5). g, Kaplan-Meier survival curves of experimental mice over time (n = 5). Note that the data for the Vehicle and CAR-iNKT + iMRAS groups were also presented in the main Fig. 6b-f. a and d, Created in BioRender. Liu, Z. (2025) https://BioRender.com/jnuo8z1 Data are presented as the mean ± s.e.m. NS, not significant; *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, by one-way ANOVA (c), or by two-way ANOVA (b and f). In all figures, n represents the number of individual mice. Source data are provided as a Source Data file.

Source data

Supplementary information

Source data

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Li, YR., Nan, H., Liu, Z. et al. Engineering an in vivo charging station for CAR-redirected invariant natural killer T cells to enhance cancer therapy. Nat. Biomed. Eng (2026). https://doi.org/10.1038/s41551-026-01629-3

Download citation

  • Received:

  • Accepted:

  • Published:

  • Version of record:

  • DOI: https://doi.org/10.1038/s41551-026-01629-3

Related Posts