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In vivo molecular imaging for immunotherapy using ultra-bright near-infrared-IIb rare-earth nanoparticles

Nature Biotechnology volume 37, pages 1322–1331 (2019)Cite this article

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Abstract

The near-infrared-IIb (NIR-IIb) (1,500–1,700 nm) window is ideal for deep-tissue optical imaging in mammals, but lacks bright and biocompatible probes. Here, we developed biocompatible cubic-phase (α-phase) erbium-based rare-earth nanoparticles (ErNPs) exhibiting bright downconversion luminescence at ~1,600 nm for dynamic imaging of cancer immunotherapy in mice. We used ErNPs functionalized with cross-linked hydrophilic polymer layers attached to anti-PD-L1 (programmed cell death-1 ligand-1) antibody for molecular imaging of PD-L1 in a mouse model of colon cancer and achieved tumor-to-normal tissue signal ratios of ~40. The long luminescence lifetime of ErNPs (~4.6 ms) enabled simultaneous imaging of ErNPs and lead sulfide quantum dots emitting in the same ~1,600 nm window. In vivo NIR-IIb molecular imaging of PD-L1 and CD8 revealed cytotoxic T lymphocytes in the tumor microenvironment in response to immunotherapy, and altered CD8 signals in tumor and spleen due to immune activation. The cross-linked functionalization layer facilitated 90% ErNP excretion within 2 weeks without detectable toxicity in mice.

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Fig. 1: Ultra-bright ~1,550 nm NIR-IIb luminescence of Zn-doped α-ErNPs.
Fig. 2: Biocompatible, rapid-excretable α-ErNPs for real-time NIR-IIb imaging under low-power LED excitation.
Fig. 3: ErNPs-aPDL1 mAb complex for in vivo PD-L1 molecular imaging and immunotherapy.
Fig. 4: In vivo two-plex NIR-IIb molecular imaging of immune responses using ErNPs-aPDL1 and PbS-aCD8 at the same ~1,600 nm emission range.
Fig. 5: In vivo two-plex NIR-IIb molecular imaging of PDL1 and CD8+ CTLs for assessing immune activation and responses.

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Data availability

The data that support the findings of this study are available from the corresponding author upon reasonable request.

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Acknowledgements

This study was supported by the National Institutes of Health (grant no. DP1-NS-105737).

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  1. These authors contributed equally: Yeteng Zhong, Zhuoran Ma, Feifei Wang.

Authors and Affiliations

  1. Department of Chemistry and Bio-X, Stanford University, Stanford, CA, USA

    Yeteng Zhong, Zhuoran Ma, Feifei Wang, Yulai Liu, Jiachen Li, Haotian Du, Mingxi Zhang, Shoujun Zhu, Qinchao Sun, Hao Wan, Ye Tian, Qiang Liu, Weizhi Wang & Hongjie Dai

  2. Key Laboratory of Luminescence and Optical Information, Ministry of Education, Department of Physics, School of Science, Beijing Jiaotong University, Beijing, China

    Xi Wang, Yijun Yang & Qiuhong Cui

  3. Departments of Molecular and Cellular Physiology and Structural Biology, Stanford University School of Medicine, Stanford, CA, USA

    Xiang Zhao & K. Christopher Garcia

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Contributions

H. Dai and Y.Z. conceived and designed the experiments. Y.Z., Z.M., F.W., X.W., Y.Y., Y.L., X.Z. and J.L. performed the experiments. Y.Z., Z.M., F.W., X.W., Y.Y., Y.L., X.Z., J.L., H. Du, M.Z., Q.C., S.Z., Q.S., H.W., Y.T., Q.L., W.W., K.C.G. and H. Dai analyzed the data and wrote the manuscript. All authors discussed the results and commented on the manuscript.

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Correspondence to Hongjie Dai.

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Supplementary information

Supplementary Information

Supplementary Figs. 1–26 and Table 1.

Reporting Summary

Supplementary Video 1

Real-time NIR-IIb in vivo dynamic imaging of ErNP-labeled cerebral blood flow in a BALB/c mouse brain on 970 nm LED lamp excitation (15 mW cm−2). Total video time: 33.2 s, frame rate: 30 fps. Similar results for n > 3 independent experiments.

Supplementary Video 2

Real-time NIR-IIb in vivo dynamic imaging distinguished the vessels in mouse scalp and under the skull. Similar results for n > 3 independent experiments.

Supplementary Video 3

Ultra-fast NIR-IIb in vivo dynamic imaging of ErNP-labeled hindlimb blood flow in a BALB/c mouse on 980 nm diode laser excitation (100 mW cm−2). Total video time: 13.2 s, frame rate: 90 fps. Similar results for n > 3 independent experiments.

Supplementary Video 4

In vivo two-plex rotation imaging of a CT-26 tumor mouse intravenously injected with ErNPs-aPDL1 and PbS-aCD8 at 0 h, 24 h and 48 h post injection. Similar results for n = 3 independent experiments

Supplementary Video 5

In vivo rotation imaging of a CT-26 tumor mouse intravenously injected with PbS-aCD8 at 24 h post injection. Similar results for n = 3 independent experiments.

Supplementary Video 6

In vivo two-plex rotation imaging of a 4T1 tumor mouse intravenously injected with ErNPs-aPDL1 and PbS-aCD8 at 24 h post injection. Similar results for n = 3 independent experiments.

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Zhong, Y., Ma, Z., Wang, F. et al. In vivo molecular imaging for immunotherapy using ultra-bright near-infrared-IIb rare-earth nanoparticles. Nat Biotechnol 37, 1322–1331 (2019). https://doi.org/10.1038/s41587-019-0262-4

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