Abstract
Modern medicine seeks precision targeting, imaging and therapy to maximize efficacy and avoid toxicities. Nanoparticles (NPs) have tremendous yet unmet clinical potential to carry and deliver imaging and therapeutic agents systemically with tissue precision. But their size contributes to rapid scavenging by the reticuloendothelial system and poor penetration of key endothelial cell (EC) barriers, limiting target tissue uptake, safety and efficacy. Here we discover the ability of the EC caveolae pumping system to outpace scavenging and deliver NPs rapidly and specifically into the lungs. Gold and dendritic NPs are conjugated to antibodies targeting caveolae of the lung microvascular endothelium. SPECT-CT imaging and biodistribution analyses reveal that rat lungs extract most of the intravenous dose within minutes to achieve precision lung imaging and targeting with high lung concentrations exceeding peak blood levels. These results reveal how much ECs can both limit and promote tissue penetration of NPs and the power and size-dependent limitations of the caveolae pumping system. This study provides a new retargeting paradigm for NPs to avoid reticuloendothelial system uptake and achieve rapid precision nanodelivery for future diagnostic and therapeutic applications.
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The data supporting the findings of this study are available within the article, Supplementary Information and Source Data files. Other relevant data are available for research purposes from the corresponding authors upon request. Source data are provided with this paper.
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Acknowledgements
We thank C. Ditterich for assistance with paper writing. We thank nanoComposix for help with TEM. This study was supported by the National Institutes of Health (https://www.nih.gov/grantsfunding) through grants awarded to J.E.S. (P01HL119165, R01CA169644 and R01HL169760). The funders had no role in study design, data collection and analysis, decision to publish or preparation of the paper.
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J.E.S. conceived the study. T.R.N., A.C., M.D.L., B.O. and J.E.S. designed the study. R.Y. and B.C. conducted antibody expression, purification, ELISA and western blotting. T.B. performed site-directed mutagenesis. A.C. prepared and performed SPECT-CT imaging and biodistribution studies of radioimmunoconjugates of PAMAM dendrimers and dendrons. O.C.-R. characterized and purified starting material antibodies. T.R.N. prepared GNP nanoimmunoconjugates. T.R.N. and O.C.-R. characterized GNP nanoimmunoconjugates. T.R.N. characterized GNP radio-nanoimmunoconjugates. T.R.N., A.C. and J.V. performed in vivo SPECT-CT imaging. A.C. and T.R.N. conducted the ex vivo biodistribution study. J.V. and T.R.N. conducted the ICP-MS study. T.R.N. and A.C. collected all data. J.K. conducted statistical analyses. T.R.N., A.C. and J.E.S. wrote the paper. M.D.L. and B.O. revised the paper. All authors read and edited the paper.
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Extended data
Extended Data Fig. 1 Functionalization of PAMAM G5 dendrimers with APP2 Fab to form supramolecular nanoassembly.
PAMAM G5 were biotinylated using NHS-PEG4-biotin then radiolabeled with N-succinimidyl-3-(4-hydroxy-3-[125I]iodophenyl)propionate and terminated with glycidol. To form supramolecular nanoassembly, the biotinylated 125I labeled PAMAM G5 dendrimers were then noncovalently functionalized with trivalent APP2 streptabodies via streptavidin (SAV)-biotin linkage.
Extended Data Fig. 2 Characterization of Gold Nanoparticles (GNPs).
Size exclusion chromatography (SEC-UV) of unconjugated antibody, GNP and GNP immunoconjugates as indicated in color legend. The UV traces (λabs = 280 nm) show high purity of the samples and absence of high and low molecular weight contaminants.
Extended Data Fig. 3 Functionalization of gold nanoparticles with radioiodinated mAPP2.
PEG-Carboxilic acid terminated gold nanopartilces were conjugated to mAPP2 antibody as described in Methods. EDC: 1-ethyl-3-(3-dimethylamino) propyl carbodiimide, NHS: N-hydroxysulfosuccinimide.
Extended Data Fig. 4 Western blot validation of antibody specificity towards rat APP2.
Membranes, containing (1) 50 ng recombinant rat APP2 protein (aa2-648 + 6XHIS), (2) 30 μg rat lung homogenate, and (3) 30 μg purified plasma membrane fraction of rat lung-derived endothelial cells, probed with mAPP2 and non-specific, mutated mAPP2X show distinct difference in target recognition. mAPP2X shows no binding to rat APP2 nor any native rat lung proteins.
Source data
Source Data Fig. 1
Biodistribution data for Fig. 1.
Source Data Fig. 3
ROI analysis data for Fig. 3.
Source Data Fig. 4
Biodistribution data for Fig. 4.
Source Data Fig. 5
Target indices data for Fig. 5.
Source Data Fig. 6
Biodistribution and targeting indices for Figs. 4–6 and ICP-MS data for Fig. 6.
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Nayak, T.R., Chrastina, A., Valencia, J. et al. Rapid precision targeting of nanoparticles to lung via caveolae pumping system in endothelium. Nat. Nanotechnol. 20, 144–155 (2025). https://doi.org/10.1038/s41565-024-01786-z
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DOI: https://doi.org/10.1038/s41565-024-01786-z
