Supramolecular Organization Predicts Protein Nanoparticle Delivery to Neutrophils for Acute Lung Inflammation Diagnosis and Treatment

Jacob W. Myerson; Priyal N. Patel; Nahal Habibi; Landis R. Walsh; Yi-Wei Lee; David C. Luther; Laura T. Ferguson; Michael H. Zaleski; Marco E. Zamora; Oscar A. Marcos-Contreras; Patrick M. Glassman; Ian Johnston; Elizabeth D. Hood; Tea Shuvaeva; Jason V. Gregory; Raisa Y. Kiseleva; Jia Nong; Kathryn M. Rubey; Colin F. Greineder; Samir Mitragotri; George S. Worthen; Vincent M. Rotello; Joerg Lahann; Vladimir R. Muzykantov; Jacob S. Brenner

In contrast to NAPs, three particles (adenovirus, AAVs, and ferritin) characterized by highly symmetric arrangement of protein subunits into a protein superstructure [44] [45] [46] did not accumulate in the inflamed neutrophil-rich lungs. These three particles have evolved structures that lead to prolonged circulation or evasion of innate immunity in . CC-BY-NC-ND 4.0 International license author/funder. It is made available under a The copyright holder for this preprint (which was not peer-reviewed) is the . https://doi.org/10.1101/2020.04.15.037564 doi: bioRxiv preprint mammals. [53] [54] [55] [56] It is conceivable that neutrophils more effectively recognize less patterned and more variable protein arrangements that may better parallel the wide variety of structures presented by the staggering diversity of microbes against which neutrophils defend. 20, 35 To support our conclusions regarding supramolecular organization and neutrophil tropism, we re-engineered liposomes, particles with no intrinsic neutrophil tropism, to behave like NAPs. Protein arrangement on the surface of DBCO-IgG liposomes was predicted to recapitulate protein agglutination seen in NAPs based on hydrophobic interactions. Introduction of DBCO to IgG entails conjugation of a highly hydrophobic moiety 57 to hydrophilic residues on the IgG. Replacing DBCO with the less hydrophobic modifying group used in SATA-maleimide conjugation 58 abrogates the inflammation specificity observed with DBCO-IgG liposomes. Likewise, titrating down the amount of DBCO on the IgG, thus limiting the hydrophobic groups on the protein, also ratchets down the targeting behavior of the DBCO-IgG liposomes. Our data therefore points towards hydrophobic interactions between proteins on the liposome surface being a determinant in liposome uptake in neutrophils in the inflamed lungs. Essentially, the DBCO-IgG liposomes may reproduce the hydrophobic interaction structural motif seen in NAPs produced by protein gelation (i.e. LDNGs).