In vivo engineering of CAR natural killer cells for immunotherapy using ligand-directed lipid nanoparticles
(1) Chen Y (2) Monkovic J (3) Mughal H (4) Chen B (5) Thadhani E (6) McMullen P (7) Self A (8) Chen Y (9) Hu Y (10) Detmer S (11) Chen M (12) Lapteva A (13) Kamat R (14) Vernon A (15) Langer R (16) Anderson DG
Chen, Monkovic, and Mughal et al. used NK cell receptor-specific scFvs to generate NK cell-targeted LNPs (NtLNPs) that transfected up to 80% of circulating and tissue NK cells, with minimal off-target effects. NtLNP delivery of CAR mRNAs generated NK cells that specifically depleted B cells in healthy mice, restored AST/ALT to baseline levels in an autoimmune hepatitis model, and cleared antigen+ melanoma cells in a solid tumor model. NtLNPs targeting NKG2a boosted efficacy in solid tumor models by mitigating Qa-1b mediated immunosuppression. In vivo and ex vivo, NtLNP-generated human CAR NK cells induced regression in humanized lymphoma and T cell leukemia mouse models.
Contributed by Paula Hochman
(1) Chen Y (2) Monkovic J (3) Mughal H (4) Chen B (5) Thadhani E (6) McMullen P (7) Self A (8) Chen Y (9) Hu Y (10) Detmer S (11) Chen M (12) Lapteva A (13) Kamat R (14) Vernon A (15) Langer R (16) Anderson DG
Chen, Monkovic, and Mughal et al. used NK cell receptor-specific scFvs to generate NK cell-targeted LNPs (NtLNPs) that transfected up to 80% of circulating and tissue NK cells, with minimal off-target effects. NtLNP delivery of CAR mRNAs generated NK cells that specifically depleted B cells in healthy mice, restored AST/ALT to baseline levels in an autoimmune hepatitis model, and cleared antigen+ melanoma cells in a solid tumor model. NtLNPs targeting NKG2a boosted efficacy in solid tumor models by mitigating Qa-1b mediated immunosuppression. In vivo and ex vivo, NtLNP-generated human CAR NK cells induced regression in humanized lymphoma and T cell leukemia mouse models.
Contributed by Paula Hochman
ABSTRACT: Targeted lipid nanoparticles (LNPs) offer a nonviral approach for in vivo cell engineering but efficient programming of natural killer (NK) cells remains underexplored. Here we develop NK cell-targeted LNPs (NtLNPs) using engineered biparatopic NK cell ligands to drive receptor-specific uptake and mRNA delivery. Lead NtLNPs achieve up to ~80% NK cell transfection with minimal off-target effects in mice. Systemic delivery of chimeric antigen receptor (CAR) mRNA using NtLNPs generates CAR NK cells in vivo. CD19-targeted CAR NKs completely deplete circulating B cells in healthy mice and CD3-targeted CAR NKs eliminate nearly all autoreactive T cells in an autoimmune hepatitis model, highlighting a potential advantage over CAR T therapies for T cell-driven disease. In solid tumors, NtLNPs targeting the inhibitory checkpoint NKG2A enhance antitumor efficacy by relieving Qa-1(b)-mediated immunosuppression. Lastly, we optimized NtLNPs for in vivo engineering of human CAR NK cells, enabling regression of two humanized hematologic malignancy models and establishing NtLNPs as a modular immunotherapy system.
Author Info:
(1) David H. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA, USA. Department of Chemical Engineering, Massachusetts Institute
of Technology, Cambridge, MA, USA. (2) David H. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA, USA. Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, MA, USA. (3) David H. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA, USA. Department of Chemistry, Massachusetts Institute of Technology, Cambridge, MA, USA. (4) David H. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA, USA. Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, MA, USA. (5) David H. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA, USA. Harvard and MIT Division of Health Sciences and Technology, Massachusetts Institute of Technology, Cambridge, MA, USA. (6) David H. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA, USA. Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, MA, USA. Department of Anesthesiology, Critical Care and Pain Medicine, Boston Children's Hospital, Boston, MA, USA. (7) David H. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA, USA. Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, MA, USA. (8) David H. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA, USA. Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, MA, USA. (9) David H. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA, USA. Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, MA, USA. (10) David H. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA, USA. Department of Chemistry, Massachusetts Institute of Technology, Cambridge, MA, USA. (11) David H. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA, USA. Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, MA, USA. (12) David H. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA, USA. Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA, USA. (13) David H. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA, USA. Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA, USA. (14) David H. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA, USA. (15) David H. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA, USA. Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, MA, USA. Department of Chemistry, Massachusetts Institute of Technology, Cambridge, MA, USA. Harvard and MIT Division of Health Sciences and Technology, Massachusetts Institute of Technology, Cambridge, MA, USA. Department of Anesthesiology, Critical Care and Pain Medicine, Boston Children's Hospital, Boston, MA, USA. Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA, USA. Institute for Medical Engineering and Science, Massachusetts Institute of Technology, Cambridge, MA, USA. (16) David H. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA, USA. dgander@mit.edu. Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, MA, USA. dgander@mit.edu. Harvard and MIT Division of Health Sciences and Technology, Massachusetts Institute of Technology, Cambridge, MA, USA. dgander@mit.edu. Department of Anesthesiology, Critical Care and Pain Medicine, Boston Children's Hospital, Boston, MA, USA. dgander@mit.edu. Institute for Medical Engineering and Science, Massachusetts Institute of Technology, Cambridge, MA, USA. dgander@mit.edu.
Citation: Nat Biotechnol 2026 Oct 13 Epub10/13/2026
