Biology subjects

Werfel, T. A.

Publications and source records attributed to Werfel, T. A..

3 recordsLinked to original sources

bioRxiv2025

Precision control of nanoparticle delivery with engineered biomimetic protein coronas

Nanoparticle delivery to tumors remains inefficient, with current nanomedicines achieving only 0.7% injected dose per gram (ID/g) of tumor tissue due to uncontrolled protein corona formation that redirects nanoparticles away from target sites. We engineered biomimetic protein coronas to control nanoparticle-protein interactions and enhance tumor targeting. Competitive binding studies using NMR spectroscopy revealed that transferrin (Tf) and fibronectin (Fn) outcompete albumin (BSA) and immunoglobulin G (IgG) for 15 nm gold nanoparticle surfaces, establishing a binding hierarchy that enables predictable corona composition. Pre-coating nanoparticles with a four-protein combination (BSA+Tf+Fn+IgG) created coronas that selectively enhanced cancer cell uptake while reducing macrophage recognition in vitro. When administered to tumor-bearing mice, these engineered coronas achieved 13 ppm/g tumor accumulation--equivalent to 4% ID/g--representing 6.5-fold improvement over bare nanoparticles and 2.6-fold improvement over PEGylated formulations. Proteomics analysis of secondary coronas formed in human serum revealed that engineered nanoparticles selectively recruit transport and adhesion proteins while limiting immune recognition signatures. The pre-formed coronas maintained targeting protein retention and reduced complement binding compared to controls. Circular dichroism confirmed minimal protein structural perturbation, preserving receptor-binding functionality for active targeting. The strategy harnesses natural protein adsorption processes to create "smart" biological interfaces that simultaneously evade immune clearance and promote tumor cell recognition through multiple receptor pathways. This approach demonstrates the feasibility of treating the coron as a programmable interface, addressing delivery limitations that have hindered clinical translation of cancer nanomedicines. For Table of Contents Only O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=109 SRC="FIGDIR/small/693224v1_ufig1.gif" ALT="Figure 1"> View larger version (61K): org.highwire.dtl.DTLVardef@5dd106org.highwire.dtl.DTLVardef@1460e3corg.highwire.dtl.DTLVardef@50c251org.highwire.dtl.DTLVardef@5604f_HPS_FORMAT_FIGEXP M_FIG C_FIG

biophysics↗

bioRxiv2024

Polymeric Micellar Nanoparticles Enable Image-guided Drug Delivery in Solid Tumors

We report the development of a nanotechnology to co-deliver chemocoxib A with a reactive oxygen species (ROS)-activatable and COX-2 targeted pro-fluorescent probe, fluorocoxib Q (FQ) enabling real time visualization of COX-2 and CA drug delivery into solid cancers, using a di-block PPS135-b-POEGA17 copolymer, selected for its intrinsic responsiveness to elevated reactive oxygen species (ROS), a key trait of the tumor microenvironment. FQ and CA were synthesized independently, then co-encapsulated within micellar PPS135-b-POEGA17 co-polymeric nanoparticles (FQ-CA-NPs), and were assessed for cargo concentration, hydrodynamic diameter, zeta potential, and ROS-dependent cargo release. The uptake of FQ-CA-NPs in mouse mammary cancer cells and cargo release was assessed by fluorescence microscopy. Intravenous delivery of FQ-CA-NPs to mice harboring orthotopic mammary tumors, followed by vital optimal imaging, was used to assess delivery to tumors in vivo. The CA-FQ-NPs exhibited a hydrodynamic diameter of 109.2 {+/-} 4.1 nm and a zeta potential ({zeta}) of -1.59 {+/-} 0.3 mV. Fluorescence microscopy showed ROS-dependent cargo release by FQ-CA-NPs in 4T1 cells, decreasing growth of 4T1 breast cancer cells, but not affecting growth of primary human mammary epithelial cells (HMECs). NP-derived fluorescence was detected in mammary tumors, but not in healthy organs. Tumor LC-MS/MS analysis identified both CA (2.38 nmol/g tumor tissue) and FQ (0.115 nmol/g tumor tissue), confirming the FQ-mediated image guidance of CA delivery in solid tumors. Thus, co-encapsulation of FQ and CA into micellar nanoparticles (FQ-CA-NPs) enabled ROS-sensitive drug release and COX-2-targeted visualization of solid tumors.

bioengineering↗

bioRxiv2023

Near-Infrared Photothermal Ablation of Biofilms using Protein-Functionalized Gold Nanospheres with a Tunable Temperature Response

Temperature-responsive nanostructures with high antimicrobial efficacy are attractive for therapeutic applications against multi-drug-resistant bacteria. Here, we report temperature-responsive nanospheres (TRNs) that are engineered to undergo self-association and agglomeration above a tunable transition temperature (Tt). Temperature-responsive behavior of the nanoparticles is obtained by functionalizing citrate-capped, spherical gold nanoparticles (AuNPs) with elastin-like polypeptides (ELPs). Using protein design principles, we achieve a broad range of attainable Tt values and photothermal conversion efficiencies ({eta}). Two approaches were used to adjust this range: First, by altering the position of the cysteine residue used to attach ELP to the AuNP, we attained a Tt range from 34-42 {degrees}C. Then, functionalizing the AuNP with an additional small globular protein, we were able to extend this range to 34-50 {degrees}C. Under near-infrared (NIR) light exposure, all TRNs exhibited reversible agglomeration. Moreover, they showed enhanced photothermal conversion efficiency in their agglomerated state relative to the dispersed state. Despite their spherical shape, TRNs have a photothermal conversion efficiency approaching that of gold nanorods ({eta} = 68{+/-}6%), yet unlike nanorods, the synthesis of TRNs requires no cytotoxic compounds. Finally, we tested TRNs for photothermal ablation of biofilms. Above Tt, NIR irradiation of TRNs resulted in a 10,000-fold improvement in killing efficiency compared to untreated controls (p < 0.0001). Below Tt, no enhanced anti-biofilm effect was observed. In conclusion, engineering the interactions between proteins and nanoparticles enables the tunable control of TRNs, resulting in a novel, anti-biofilm nanomaterial with low cytotoxicity. TOC Image O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=98 SRC="FIGDIR/small/553096v1_ufig1.gif" ALT="Figure 1"> View larger version (65K): org.highwire.dtl.DTLVardef@ffb632org.highwire.dtl.DTLVardef@12cbb4org.highwire.dtl.DTLVardef@1fbac00org.highwire.dtl.DTLVardef@1d950b6_HPS_FORMAT_FIGEXP M_FIG C_FIG

bioengineering↗