CHINESE JOURNAL OF DRUG EVALUATION >

2025 , Vol. 42 >Issue 6: 409 - 409-415

DOI: https://doi.org/10.2095-3593.2025.120001

Research Progress and General considerations on Nonclinical Evaluation of In Vivo Chimeric Antigen Receptor (CAR)-T Cell Therapy

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  • Center For Drug Evaluation， NMPA， Beijing 100022， China

Received date: 2025-11-26

  Accepted date: 2026-01-07

  Online published: 2026-01-07

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Abstract

Chimeric antigen receptor (CAR)-T cell therapy has demonstrated significant therapeutic potential in hematologic malignancies (particularly B-cell malignancies). However， its broad application faces considerable challenges， including complex ex vivo manufacturing processes and high production costs. In recent years， with rapid advancements in RNA therapeutics and targeted delivery systems， in vivo generation of chimeric antigen receptor T cells (in vivo CAR-T) has emerged as an innovative strategy. This approach utilizes viral vectors or lipid nanoparticles (LNPs) as targeted delivery systems to directly introduce genetic material encoding the CAR into patients， enabling in vivo T cell engineering. This strategy fundamentally eliminates the need for cumbersome ex vivo cell manipulation steps and traditional chemotherapy-based preconditioning regimens. This article systematically reviews the technological progress and nonclinical research considerations for in vivo CAR-T. As in vivo CAR-T combines attributes of both gene therapy and cell therapy， involves diverse delivery vectors， and exhibits varied types and complex mechanisms， its nonclinical studies can be guided by risk-based， case-by-case principles. Within the framework of existing relevant technical guidelines， scientifically sound nonclinical studies should be rationally designed and conducted to generate robust experimental data supporting clinical trials and marketing approval.

Key words： ; font-size:medium; ">In vivo CAR-T； Non-clinical research； Viral vector； LNP vector

Cite this article

ZHOU Yu, ZHANG Min, YAN Liping . Research Progress and General considerations on Nonclinical Evaluation of In Vivo Chimeric Antigen Receptor (CAR)-T Cell Therapy[J]. CHINESE JOURNAL OF DRUG EVALUATION, 2025 , 42(6) : 409 -409-415 . DOI: 10.2095-3593.2025.120001

References

 [1] Cappell KM， Kochenderfer JN. Long-term outcomes following CAR T cell therapy：what we know so far[J]. Nat Rev Clin Oncol， 2023， 20(6)： 359-371.
     [2] Mikhael J， Fowler J， Shah N. Chimeric antigen receptor T-cell therapies： barriers and solutions to access[J]. JCO Oncol Pract， 2022， 18(12)：800-807.
     [3] Sterner RC， Sterner RM. CAR-T cell therapy： current limitations and potential strategies[J]. Blood Cancer J， 2021， 11(4)： 69.
     [4] Lickefett B， Chu L， Ortiz-Maldonado V， et al. Lymphodepletion—an essential but undervalued part of the chimeric antigen receptor T-cell therapy cycle[J]. Front Immunol， 2023， 14：1303935.
     [5] Zhou Q， Schneider IC， Edes I， et al. T-cell receptor gene transfer exclusively to human CD8+ cells enhances tumor cell killing[J]. Blood， 2012， 120(22)： 4334-4342.
     [6] Anliker B， Abel T， Kneissl S， et al. Specific gene transfer to neurons， endothelial cells and hematopoietic progenitors with lentiviral vectors[J]. Nat Methods， 2010， 7(11)： 929-935.
     [7] Ho N， Agarwal S， Milani M， et al. In vivo generation of CAR T cells in the presence of human myeloid cells[J]. Mol Ther Methods Clin， 2022， 26：144-156.
     [8] Andorko JI， Russell RM， Schnepp BC， et al. Targeted in vivo delivery of genetic medicines utilizing an engineered lentiviral vector platform results in CAR T and NK cell generation[J]. Mol Ther， 2025， 33(10)：4937-4952.
     [9] Russell RM， Boral D， Andorko JI， et al. Rational design of a detargeted vesiculovirus fusogen to enable targeted in vivo gene delivery[EB/OL]. （2024-05-07）[2025-10-25]. https：//doi. org/10. 1016/j. ymthe.
     [10] Mullen KF. In vivo generation of both CAR T cells and CAR NK cells using a CD7 targeted lentiviral vector[EB/OL]. (2024-05-07) [2025-10-25]. https：//doi. org/10. 1016/j. ymthe.
     [11] Nicolai CJ， Parker MH， Qin J， et al. In vivo CAR T-cell generation in nonhuman primates using lentiviral vectors displaying a multidomain fusion ligand[J]. Blood， 2024， 144(9)： 977-987.
     [12] Michels KR， Sheih A， Hernandez SA， et al. Preclinical proof of concept for VivoVec， a lentiviral-based platform for in vivo CAR T-cell engineering[J]. J Immunother Cancer， 2023， 11(3)：e006292.
     [13] Xu J， Liu L， Parone P， et al. In-vivo B-cell maturation antigen CAR T-cell therapy for relapsed or refractory multiple myeloma[J]. Lancet， 2025， 406(10500)： 228-231.
     [14] Wolff JA， Malone RW， Williams P， et al. Direct gene transfer into mouse muscle in vivo[J]. Science， 1990， 247(4949 Pt1)： 1465-1468.
     [15] Wang Y， Zhao H， Cochran E， et al. In vivo programming of myeloid cells by mRNA mediated delivery of novel FcA fusion receptor activates anti-tumor immunity[EB/OL]. (2022-10-07) [2025-10-25]. https：//doi. org/10. 1136/jitc-2022-SITC2022.
     [16] Franz AM， Riener R， Wang A， et al. Profound B Cell depletion and repopulation with predominantly na?ve B cells in non-human primates achieved through a novel in vivo CD8-targeted lipid nanoparticle mRNA CAR[EB/OL]. (2024-11-14) [2025-10-25]. https：//rheumatology. org/annual-meeting.
     [17] Hunter TL， Bao Y， Zhang Y， et al. In vivo CAR T cell generation for cancer and autoimmune disease[J]. Science， 2025， 388(6753)： 1311-1317.
     [18] Wesselhoeft RA， Kowalski PS， Anderson DG. Engineering circular RNA for potent and stable translation in eukaryotic cells[J]. Nat Commun， 2018， 9(1)： 2629.
     [19] Wesselhoeft RA， Kowalski PS， Parker-Hale FC， et al. RNA circularization diminishes immunogenicity and can extend translation duration in vivo[J]. Mol Cell， 2019， 74(3)： 508-520.
     [20] Cunningham J. In vivo delivery of a CD20 CAR using a CD8-targeted fusosome in southern pig-tail macaques (M. nemestrina) results in B cell depletion [EB/OL]. (2021-12-11) [2025-10-25]. https：//www. hematology. org.
     [21] Mullard A. In vivo CAR T cells move into clinical trials[J]. Nat Rev Drug Discov， 2024， 23(10)： 727-730.
     [22] Wang Y， Wang X， Pan Q， et al. CircRNA-based， non-integrated， in vivo panCAR-mediated B cell immune resetting in mouse models and non-human primates[C/OL]. bioRxiv. (2025-08-11) [2025-10-25]. https：//doi. org/10. 1101/2025. 08. 11. 669448.
     [23] Wang Y， Lin L， Wang X， et al. Synergically enhanced anti-tumor immunity of in vivo CAR by circRNA vaccine boosting[C/OL]. bioRxiv. (2024-07-05) [2025-10-25]. https：//doi. org/10. 1101/2024. 07. 05. 600312.
     [24] Nicolai C， Qin J， Pankau M， et al. VivoVec： a novel lentiviral-based in vivo CAR T cell generation platform with viral particle surface engineering incorporating T cell activating and co-stimulatory ligands[EB/OL]. (2022-05-16) [2025-10-25]. https：//www. umoja-biopharma. com/wp-content/uploads/2022/11/CN-ASGCT-Poster-Umoja1.
     [25] Brandes AH， Koo ASM， Perez A， et al. Nonclinical Toxicology， Biodistribution， and Pharmacokinetics of UB-VV111， an In Vivo Anti-CD19 CAR T Cell Therapy[EB/OL]. （2024-10-05） [2025-10-25]. https：//doi. org/10. 1182/blood-2024-211032.
     [26] Wesselhoeft RA， Kowalski PS， Parker-Hale FC， et al. RNA circularization diminishes immunogenicity and can extend translation duration in vivo[J]. Mol Cell， 2019， 74(3)： 508-520.
     [27] Contrastano SG. Potent in vivo transduction by iGPS particles generates CAR T cells with durable anti-tumor activity[EB/OL]. （2023-05-17）[2025-10-25]. https：//keloniatx. com/wp-content/uploads/2023/05/2023-ASGCT-Kelonia-Presentation.
     [28] Beura ET， Latimer HJ， Wong D， et al. T cell-specific in vivo transduction with preclinical candidate KLN-1010 generates BCMA directed CAR T cells with potent anti-multiple myeloma activity[EB/OL]. （2023-03-22） [2025-10-25]. https：//doi. org/10. 1158/1538-7445.
     [29] Hu J， Dunbar CE. T-cell lymphomas in recipients of CAR-T cells： assessing risks and causalities[J]. Blood， 2024， 144(24)： 2473-2481.
     [30] Hamilton MP， Sugio T， Noordenbos T， et al. Risk of second tumors and T-cell lymphoma after CAR T-cell therapy[J]. N Engl J Med， 2024， 390(22)： 2047-2060.
     [31] Ruella M， June CH. CAR T-cell resistance to oncogenic transformation[J]. Blood Cancer Discov， 2024， 5(4)： 229-233.
     [32] Xiao T， Zha G， Shi L， et al. An vivo CAR T producer (HN2301) for autoimmune disease enables B cell depletion in NHP tissues and performs therapeutic effect in an SLE mouse model[EB/OL]. （2025-05-13） [2025-10-25]. https：//www. sciencedirect. com/journal/molecular-therapy/vol/33/issue/4/suppl/S1.
     [33] Wang Q， Xiao Z， Zheng X， et al. In vivo CD19 CAR T-cell therapy for refractory systemic lupus erythematosus[J]. N Engl J Med， 2025， 393(15)：1542-1544.
     [34] Muraro PA， Martin R， Mancardi GL， et al. Autologous haematopoietic stem cell transplantation for treatment of multiple sclerosis[J]. Nat Rev Neurol， 2017， 13(7)： 391-405.
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