A pinboard by
Kyle Isaacson

Graduate Researcher, University of Utah


Loading cancer drugs into nanogels for improved tumor-targeting can improve current treatments

Breast cancer is a debilitating disease that will affect 1 out of every 8 women. Current chemotherapy treatments involve the injection of highly toxic compounds through patients' bloodstreams to destroy disseminated cancer cells. Unfortunately, these toxic compounds often devastate healthy tissues and bring about a plethora of side effects. Nanoparticles are often proposed to help improve the transport of chemotherapeutics through the blood, but researchers rarely develop methods that inhibit the passage of such nanoparticle drug delivery systems into healthy tissues.

My research focuses on the development of a completely protein-based nanogel that is too large to readily leave the bloodstream and enter into tissues normally, a process called extravasation. However, the nanogels' elastin units respond to temperature and shrivel when heated. This shriveling causes the chemotherapeutic-carrying nanogels to shrink approximately 50% in size, allowing them to freely extravasate from the bloodstream. Tumors are identified and heated slightly above body temperature, allowing the nanogels to only extravasate at tumor sites. This localized extravasation concentrates the toxic chemotherapeutic drugs near cancer cells and reduces contact with healthy cells, effectively improving treatment efficacy and substantially reducing side effects.


On the scattered light by dilute aqueous dispersions of nanogel particles.

Abstract: This work deals with the scattered light by nanoparticles formed by a temperature sensitive polymer networks, namely nanogel particles. The scattered light is measured as a function of the scattering angle at temperatures below and above the volume phase transition temperature (VPTT) of nanogel particles. Our experimental results indicate that nanogel particles have a core-shell structure, formed by a uniform highly cross-linked core surrounded by a fuzzy shell where the polymer density decays to zero gradually for swollen configurations and sharply for shrunken states. The theoretical fitting of the experimental curves shows that the scattered light at low angle obeys a decreasing power law with the scattering vector, q(-α). The value of exponent α provides information about the radial dependence of the polymer density at the external shell of the particles for swollen nanogels, and about the degree of roughness of the surface for the case of shrunken nanogels. On the one hand, at low temperatures (below the VPPT), the nanogel particle is in the swollen state and the light scattering data show that its shell structure follows a fractal behaviour, with a polymer density that decays as r(α-3), where r is the distance to the particle centre. On the other hand, above the VPPT the results indicate that nanogel collapses into a core of uniform polymer density and a rough shell, with a fractal surface dimension of 2.5.

Pub.: 04 Apr '15, Pinned: 06 Sep '17

Competition between excluded-volume and electrostatic interactions for nanogel swelling: effects of the counterion valence and nanogel charge.

Abstract: In this work the equilibrium distribution of ions around a thermo-responsive charged nanogel particle in an electrolyte aqueous suspension is explored using coarse-grained Monte Carlo computer simulations and the Ornstein-Zernike integral equation theory. We explicitly consider the ionic size in both methods and study the interplay between electrostatic and excluded-volume effects for swollen and shrunken nanogels, monovalent and trivalent counterions, and for two different nanogel charges. We find good quantitative agreement between the ionic density profiles obtained using both methods when the excluded repulsive force exerted by the cross-linked polymer network is taken into account. For the shrunken conformation, the electrostatic repulsion between the charged groups provokes a heterogeneous polymer density profile, leading to a nanogel structure with an internal low density hole surrounded by a dense corona. The results show that the excluded-volume repulsion strongly hinders the ion permeation for shrunken nanogels, where volume exclusion is able to significantly reduce the concentration of counterions in the more dense regions of the nanogel. In general, we demonstrate that the thermosensitive behaviour of nanogels, as well as their internal structure, is strongly influenced by the valence of the counterions and also by the charge of the particles. On the one hand, an increase of the counterion valence moves the swelling transition to lower temperatures, and induces a major structuring of the charged monomers into internal and external layers around the crown for shrunken nanogels. On the other hand, increasing the particle charge shifts the swelling curve to larger values of the effective radius of the nanogel.

Pub.: 22 Feb '17, Pinned: 06 Sep '17

Protein corona formation on colloidal polymeric nanoparticles and polymeric nanogels: impact on cellular uptake, toxicity, immunogenicity and drug release properties.

Abstract: The adsorption of biomolecules to the surfaces of a nanoparticle (NP) following administration into biological environments is widely recognised. In particular, the "protein corona" is well understood in terms of formation kinetics and its impact upon the biological interactions of NPs. Its presence is an essential consideration in the design of therapeutic NPs. In the present study, the protein corona of six polymeric nanoparticles of prospective therapeutic use, were investigated. These included three colloidal NPs-soft core-multi-shell (CMS) NPs, plus solid cationic Eudragit® RS (EGRS) and anionic ethyl cellulose (EC) nanoparticles-and three nanogels (NGs)-thermoresponsive dendritic-polyglycerol (dPG) nanogels (NGs) and two amino-functionalised dPG-NGs. Following incubation with human plasma, protein coronas were characterised and their biological interactions compared with pristine NPs. All NPs demonstrated protein adsorption and increased hydrodynamic diameters, although the solid EGRS and EC NPs bound notably more protein than the other tested particles. Shifts towards moderately negative surface charges were also observed for all corona bearing NPs, despite varied zeta potentials in their pristine state. While the uptake and cellular adhesion of the colloidal NPs in primary human keratinocytes and HUVECS were significantly decreased when bearing the protein corona, no obvious impact was seen in the NGs. By contrast, corona bearing NGs induced marked increases in cytokine release from primary human macrophages not seen with corona bearing colloidal NPs. Despite this, no apparent enhancement to in vitro toxicity was noted. Finally, drug release from EGRS and EC NPs was assessed, where a decrease was seen in the EGRS NPs alone. Together these results provide a direct comparison of the physical and biological impact the protein corona has on NPs of widely varied character, and in particular highlights a distinction between the corona's effects on NGs and colloidal NPs.

Pub.: 17 May '17, Pinned: 06 Sep '17

Silk-elastinlike protein polymers enhance the efficacy of a therapeutic glycosaminoglycan for prophylactic treatment of radiation-induced proctitis.

Abstract: Radiation-induced proctitis (RIP) is the most common clinical adverse effect for patients receiving radiotherapy as part of the standard course of treatment for ovarian, prostate, colon, and bladder cancers. RIP limits radiation dosage, interrupts treatment, and lowers patients' quality of life. A prophylactic treatment that protects the gastrointestinal tract from deleterious effects of radiotherapy will significantly improve patient quality of life and may allow for higher and more regular doses of radiation therapy. Semi-synthetic glycosaminoglycan (GAG), generated from the sulfation of hyaluronic acid, are anti-inflammatory but have difficulty achieving therapeutic levels in many tissues. To enhance the delivery of GAG, we created an in situ gelling rectal delivery system using silk-elastinlike protein polymers (SELPs). Using solutions of SELP 815K (which contains 6 repeats of blocks comprised of 8 silk-like units, 15 elastin-like units, and 1 lysine-substituted elastin-like unit) with GAG GM-0111, we created an injectable delivery platform that transitioned in <5min from a liquid at room temperature to a hydrogel at body temperature. The hydrogels released 50% of their payload within 30min and enhanced the accumulation of GAG in the rectum compared to traditional enema-based delivery. Using a murine model of radiation-induced proctitis, the prophylactic delivery of a single dose of GAG from a SELP matrix administered prior to irradiation significantly reduced radiation-induced pain after 3, 7, and 21days by 53±4%, 47±10%, and 12±6%, respectively. Matrix-mediated delivery of GAG by SELP represents an innovative method for more effective treatment of RIP and promises to improve quality of life of cancer patients by allowing higher radiotherapy doses with improved safety.

Pub.: 25 Feb '17, Pinned: 25 Aug '17

Application of thermally responsive polypeptides directed against c-Myc transcriptional function for cancer therapy.

Abstract: Elastin-like polypeptides are biopolymers composed of the pentapeptide repeat Val-Pro-Gly-Xaa-Gly. Elastin-like polypeptides are soluble in aqueous solution below their transition temperature, but they hydrophobically collapse and aggregate when the temperature is raised above the transition temperature. Previous studies have suggested that the aggregation of these polypeptides in response to externally applied hyperthermia may be exploited in the use of elastin-like polypeptide for thermally targeted drug delivery. This work shows the application of elastin-like polypeptide as a delivery vehicle for a short peptide that can inhibit the transcriptional function of a specific oncogene. The coding sequence for elastin-like polypeptide was modified by the addition of the membrane translocating sequence penetratin and a peptide derived from helix 1 of the helix-loop-helix region of c-Myc (H1-S6A,F8A), known to inhibit c-Myc transcriptional function. The designed polypeptide (Pen-ELP-H1) was then expressed and purified from Escherichia coli. Cellular uptake of Pen-ELP-H1 is enhanced by both the penetratin sequence and by the hyperthermia-induced phase transition as shown by flow cytometry studies. Using immunofluorescence and reverse transcription-PCR, we show that Pen-ELP-H1 is able to disrupt the nuclear localization of c-Myc and inhibit transcriptional activation by c-Myc. Cell proliferation studies showed that Pen-ELP-H1 inhibits growth of MCF-7 cells. Furthermore, the use of hyperthermia increased the antiproliferative effect of a thermally responsive Pen-ELP-H1 approximately 2-fold compared with a nonthermally responsive control polypeptide. These studies show that genetically engineered elastin-like polypeptide carriers may provide a new way to thermally target specific oncogene inhibitors to solid tumors.

Pub.: 16 Jul '05, Pinned: 25 Aug '17

Hydrophobic drug-triggered self-assembly of nanoparticles from silk-elastin-like protein polymers for drug delivery.

Abstract: Silk-elastin-like protein polymers (SELPs) combine the mechanical and biological properties of silk and elastin. These properties have led to the development of various SELP-based materials for drug delivery. However, SELPs have rarely been developed into nanoparticles, partially due to the complicated fabrication procedures, nor assessed for potential as an anticancer drug delivery system. We have recently constructed a series of SELPs (SE8Y, S2E8Y, and S4E8Y) with various ratios of silk to elastin blocks and described their capacity to form micellar-like nanoparticles upon thermal triggering. In this study, we demonstrate that doxorubicin, a hydrophobic antitumor drug, can efficiently trigger the self-assembly of SE8Y (SELPs with silk to elastin ratio of 1:8) into uniform micellar-like nanoparticles. The drug can be loaded in the SE8Y nanoparticles with an efficiency around 6.5% (65 ng doxorubicin/μg SE8Y), S2E8Y with 6%, and S4E8Y with 4%, respectively. In vitro studies with HeLa cell lines demonstrate that the protein polymers are not cytotoxic (IC50 > 200 μg/mL), while the doxorubicin-loaded SE8Y nanoparticles showed a 1.8-fold higher cytotoxicity than the free drug. Confocal laser scanning microscopy (CLSM) and flow cytometry indicate significant uptake of the SE8Y nanoparticles by the cells and suggest internalization of the nanoparticles through endocytosis. This study provides an all-aqueous, facile method to prepare nanoscale, drug-loaded SELPs packages with potential for tumor cell treatments.

Pub.: 18 Feb '14, Pinned: 25 Aug '17