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Subcutaneous biodegradable scaffolds for restimulating the antitumour activity of pre-administered CAR-T cells

Abstract

The efficacy of adoptive T-cell therapies based on chimaeric antigen receptors (CARs) is limited by the poor proliferation and persistence of the engineered T cells. Here we show that a subcutaneously injected biodegradable scaffold that facilitates the infiltration and egress of specific T-cell subpopulations, which forms a microenvironment mimicking features of physiological T-cell activation, enhances the antitumour activity of pre-administered CAR-T cells. CAR-T-cell expansion, differentiation and cytotoxicity were driven by the scaffold’s incorporation of co-stimulatory bound ligands and soluble molecules, and depended on the types of co-stimulatory molecules and the context in which they were presented. In mice with aggressive lymphoma, a single, local injection of the scaffold following non-curative CAR-T-cell dosing led to more persistent memory-like T cells and extended animal survival. Injectable biomaterials with optimized ligand presentation may boost the therapeutic performance of CAR-T-cell therapies.

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Fig. 1: TES enable CAR-T-cell restimulation in vitro.
Fig. 2: Subcutaneously injected TES are biodegradable and remain functional.
Fig. 3: Injected TES facilitate T-cell infiltration, activation and egress.
Fig. 4: A single local injection of TES enhances systemic antitumour activity in an aggressive liquid tumour model.

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Data availability

The data supporting the findings in this study are available within the paper and its Supplementary Information. All data generated in this study are available from the corresponding author on reasonable request. Source data are provided with this paper.

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Acknowledgements

We thank A. Najibi for critical reading of the manuscript and D. Neuberg for statistical guidance; A. Cheung, O. Ali, Z. Good, N. Cerri, E. Smith, C. Louvet, C. Wu, M. Pezzone, E. Doherty, H. Ijaz, C. Johnson, D. White, M. Perez, E. Zigon, G. Cuneo and T. Ferrante for many important technical and scientific discussions. This work was supported by the Wyss Institute at Harvard University (D.J.M.), the Food and Drug Administration (grant number 5R01FD006589), and the National Institutes of Health under grant numbers R01 EB015498, R01 CA276459, and U01 CA214369. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. D.K.Y.Z. acknowledges support from the Canadian Institutes of Health Research (CIHR). J.M.B. acknowledges support from the National Cancer Institute (grant number K00CA234959). Material characterization was performed at the Center for Nanoscale Systems (CNS) at Harvard University, which is supported by the National Science Foundation (grant number 1541979).

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Authors

Contributions

D.K.Y.Z., J.M.B. and D.J.M. conceived the project. D.K.Y.Z., J.M.B., K.A.-B., Y.L., Y.B., I.d.L., M.C.S. and R.T. performed experiments. D.K.Y.Z. and J.M.B. analysed data and generated figures. D.K.Y.Z., J.M.B. and D.J.M. wrote the paper. All authors commented on the paper.

Corresponding author

Correspondence to David J. Mooney.

Ethics declarations

Competing interests

D.J.M. is an inventor in patent applications Harvard University has filed in relation to the TES technology, which has been licensed to Lyell Immunopharma (US Patent App. 18/072,449). D.J.M. has stock in Lyell Immunopharma. D.J.M, D.K.Y.Z. and J.M.B. are listed as inventors in a patent application based on the findings of this study (WO 2023/147185). The other authors declare no competing interests.

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Nature Biomedical Engineering thanks Xiaoyuan Chen, Zhen Gu and Jonathan Schneck for their contribution to the peer review of this work.

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Extended data

Extended Data Fig. 1 TES dictates T-cell proliferation kinetics in vivo.

(a) Naïve, unstimulated, primary human T cells were labeled with CFSE and co-delivered subcutaneously with TES presenting αCD3/αCD28/IL-2 or without any ligands (labeled ‘blank’). T-cell proliferation as indicated by CFSE dilution of cells isolated from TES-containing explants at 5 days post injection. (b,c) Analysis of TES’ T-cell boosting capacity in different tumor burden contexts. Tumor-bearing animals (inoculated with 5 × 105 Raji-luc cells) were treated with either a sub-curative dose of 5 × 105 CAR-T cells or a curative dose of 1 × 106 CAR-T cells. TES were injected subcutaneously 5 days following CAR-T cell dosing, and circulating T cells in the blood were subsequently analyzed. (b) Frequency of CD3+ T cells in circulating blood. (c) Number of CD3+ T cells in the blood following subcutaneous scaffold injection under sub-curative CAR-T dosing conditions. (d, e) The capacity of injected material scaffolds to boost T-cell proliferation was evaluated in a xenograft lymphoma model. Animals were inoculated with 5 × 105 Raji-luc cells and treated 4 days afterwards with a curative dose of 1 × 106 CD19 CAR-T cells. (d) Scaffolds were injected into mice with baseline CAR-T, and human T cells were detected in αCD3/αCD28 presenting TES, while very few human T cells were detected in non-ligand-presenting ‘blank’ TES. Left: representative FACS plots showing enrichment of CD3+ T cells. (e) Expression of CD25 among T cells in day 14-explanted TES. Left: representative FACS histoplots. Data represents n = 3-4 mice per condition and mean ± s.e.m. and is representative of at least two experimental replicates. Comparisons in (d, e) were measured using unpaired Student’s T-tests with Welch’s correction for unequal s.d. Data in (b) represents mean ± s.e.m. of n = 5–7 mice.

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Zhang, D.K.Y., Brockman, J.M., Adu-Berchie, K. et al. Subcutaneous biodegradable scaffolds for restimulating the antitumour activity of pre-administered CAR-T cells. Nat. Biomed. Eng (2024). https://doi.org/10.1038/s41551-024-01216-4

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