Todd J. Harris

Systematic Surface Engineering of Magnetic Nanoworms for in vivo Tumor Targeting

In the design of nanoparticles that can target disease tissue in vivo, parameters such as targeting ligand density, type of target receptor, and nanoparticle shape can play an important role in determining the extent of accumulation. Herein, a systematic study of these parameters for the targeting of mouse xenograft tumors is performed using superparamagnetic iron oxide as a model nanoparticle system. The type of targeting peptide (recognizing cell surface versus extracellular matrix), the surface coverage of the peptide, its attachment chemistry, and the shape of the nanomaterial [elongated (nanoworm, NW) versus spherical (nanosphere, NS)] are varied. Nanoparticle circulation times and in vivo tumor-targeting efficiencies are quantified in two xenograft models of human tumors (MDA-MB-435 human carcinoma and HT1080 human fibrosarcoma). It is found that the in vivo tumor-targeting ability of the NW is superior to that of the NS, that the smaller, neutral CREKA targeting group is more effective than the larger, positively charged F3 molecule, that a maximum in tumor-targeting efficiency and blood half-life is observed with approximately 60 CREKA peptides per NW for either the HT1080 or the MDA-MB-435 tumor types, and that incorporation of a 5-kDa polyethylene glycol linker improves targeting to both tumor types relative to a short linker. It is concluded that the blood half-life of a targeting molecule-nanomaterial ensemble is a key consideration when selecting the appropriate ligand and nanoparticle chemistry for tumor targeting.

Protease-Triggered Unveiling of Bioactive Nanoparticles

Nanomaterials modified with biological recognition motifs acquire a myriad of functions that can be exploited for the diagnosis and treatment of cancer. Nevertheless, while bioactive domains can be used to target nanoparticles to cell receptors, shuttle them across cell membranes, and activate cell signaling, such modifications typically include cationic or hydrophobic regions that lead to rapid reticuloendothelial system (RES) clearance of particles from the blood, ultimately reducing particle accumulation in tumors. [1,2] Further functionalization with hydrophilic polymers like poly(ethylene glycol) (PEG) can improve blood half-lives and tumor accumulation, but often at the expense of efficient ligandmediated nanoparticle binding. [3‐5] To address this tradeoff between improved biodistribution and optimal functionality on nanoparticles, we present a general strategy for reversibly veiling bioactive domains on nanoparticles using sterically protective polymers. We demonstrate that these materials effectively accumulate via the hyperpermeable vasculature of tumors and can be activated by cancer-secreted proteases to unveil hidden functional domains.

Functional delivery of siRNA in mice using dendriworms.

Small interfering RNAs (siRNAs) mediate cleavage of specific, complementary mRNA sequences and thus regulate gene expression. Not surprisingly, their use for treatment of diseases that are rooted in aberrant gene expression, such as cancer, has become a paradigm that has gained wide interest. Here, we report the development of dendrimer-conjugated magnetofluorescent nanoworms that we call “dendriworms” as a modular platform for siRNA delivery in vivo. This platform maximizes endosomal escape to robustly produce protein target knockdown in vivo, and is tolerated well in mouse brain. We demonstrate that siRNA-carrying dendriworms can be readily internalized by cells and enable endosomal escape across a wide range of loading doses, whereas dendrimers or nanoworms alone are inefficient. Further, we show that dendriworms carrying siRNA against the epidermal growth factor receptor (EGFR) reduce protein levels of EGFR in human glioblastoma cells by 70−80%, 2.5-fold more efficiently than commercial cationic lipids. Dendriworms were well-tolerated after 7-days of convection-enhanced delivery to the mouse brain and in an EGFR-driven transgenic model of glioblastoma, anti- EGFR dendriworms led to specific and significant suppression of EGFR expression. Collectively, these data establish dendriworms as a multimodal platform that enables fluorescent tracking of siRNA delivery in vivo, cellular entry, endosomal escape, and knockdown of target proteins.

Tissue-specific gene delivery via nanoparticle coating

The use of biomaterials for gene delivery can potentially avoid many of the safety concerns with viral gene delivery. However, the efficacy of polymeric gene delivery methods is low, particularly in vivo. One significant concern is that the interior and exterior composition of polymeric gene delivery nanoparticles are often coupled, with a single polymer backbone governing all functions from biophysical properties of the polymer/DNA particle to DNA condensation and release. In this work we develop electrostatically adsorbed poly(glutamic acid)-based peptide coatings to alter the exterior composition of a core gene delivery particle and thereby affect tissue-specificity of gene delivery function in vivo. We find that with all coating formulations tested, the coatings reduce potential toxicity associated with uncoated cationic gene delivery nanoparticles following systemic injection. Particles coated with a low 2.5:1 peptide:DNA weight ratio (w/w) form large 2 micro sized particles in the presence of serum that can facilitate specific gene delivery to the liver. The same particles coated at a higher 20:1w/w form small 200nm particles in the presence of serum that can facilitate specific gene delivery to the spleen and bone marrow. Thus, variations in nanoparticle peptide coating density can alter the tissue-specificity of gene delivery in vivo.

Visual Sensor for Sterilization of Polymer Fixtures Using Embedded Mesoporous Silicon Photonic Crystals

A porous photonic crystal is integrated with a plastic medical fixture (IV connector hub) to provide a visual colorimetric sensor to indicate the presence or absence of alcohol used to sterilize the fixture. The photonic crystal is prepared in porous silicon (pSi) by electrochemical anodization of single crystal silicon, and the porosity and the stop band of the material is engineered such that the integrated device visibly changes color (green to red or blue to green) when infiltrated with alcohol. Two types of self-reporting devices are prepared and their performance compared: the first type involves heat-assisted fusion of a freestanding pSi photonic crystal to the connector end of a preformed polycarbonate hub, forming a composite where the unfilled portion of the pSi film acts as the sensor; the second involves generation of an all-polymer replica of the pSi photonic crystal by complete thermal infiltration of the pSi film and subsequent chemical dissolution of the pSi portion. Both types of sensors visibly change color when wetted with alcohol, and the color reverts to the original upon evaporation of the liquid. The sensor performance is verified using E. coli-infected samples.