Duvall Lab

Research

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The Duvall Advanced Therapeutics Laboratory specializes in design and application of smart polymer-based technologies for: (1) intracellular delivery of biological drugs such as peptides and nucleic acids, (2) proximity-activated targeting of drugs to sites of inflammation and matrix remodeling, and (3) long-term, “on-demand” drug release from localized depots. These delivery systems are designed to improve the therapeutic index of existing drugs and/or to serve as enabling technologies for manipulation of intracellular targets currently considered to be “undruggable”. To achieve optimal, finely-tuned properties for these varied biomedical applications, polymers are utilized that respond to one or more environmental stimuli including pH, matrix metalloproteinases, reactive oxygen species, and temperature. Below, a sampling of topics currently under investigation in the ATL are highlighted.

 

Technologies

Disease Targets


Technologies Used & Under Development


Controlled Polymerization for Creation of Multi-Block and Functionalized Polymers

Several projects in the ATL are aimed at development of novel applications using the RAFT (reversible addition fragmentation chain transfer) polymerization technique. These projects leverage the exquisite synthetic control afforded by RAFT to create pharmaceutical grade polymer-based therapeutics with well-defined molecular weight, architecture, and chemo-selective bioconjugation sites.  Current RAFT-based projects include development of multi-functional, tumor-targeted drugs for breast cancer therapy and new technologies that can achieve environmentally-dependent and/or “cell-demanded” drug release in response to reactive oxygen species and/or enzyme activity.  We are also developing multi-block and functionalized polymer structures optimized to supra-assemble into macroscale structures such as hydrogels.
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  1. MK Gupta, JR Martin, BR Dollinger, ME Hattaway, CL Duvall. Thermogelling, ABC Triblock Copolymer Platform for Resorbable Hydrogels with Tunable, Degradation-Mediated Drug Release. Advanced Functional Materials. 2017 (Dec 15), 27, 1704107.
  2. MA Jackson, TA Werfel, E Curvino, F Yu, TE Kavanaugh, SM Sarett, M Dockery, KV Kilchrist, A Jackson, TD Giorgio, CL Duvall. Zwitterionic Nanocarrier Surface Chemistry Improves siRNA Tumor Delivery and Silencing Activity Relative to Polyethylene Glycol. ACS Nano. 2017 Jun 27;11(6):5680-5696.
  3. H Li, SS Yu, M Miteva,CE Nelson, T Werfel, TD Giorgio, CL Duvall. Matrix Metalloproteinase Responsive, Proximity-activated Targeting Polymeric Nanoparticles for siRNA Delivery.  Advanced Functional Materials. June 25, 2013 23: 3040–3052. doi: 10.1002/adfm.201202215

 


Tissue Regenerative Biomaterial Scaffolds

Researchers in the ATL have developed novel, biodegradable materials that are broken down by cell-generated reactive oxygen species (ROS). This method of material degradation matches the rate of new tissue in-growth in vivo, optimizing tissue repair/regeneration. These materials include injectable poly(thioketal-urethane) (PTK-UR) scaffolds that form porous constructs after delivery to a wound site, and thermally-responsive PPS-based hydrogels that are liquids at room temperature but form stable gels at body temperature. These tissue regenerative materials can be used in many applications, including wound healing, sustained local drug delivery, and therapeutic cell delivery.
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  1. BR Dollinger, MK Gupta, JR Martin, CL Duvall.  ROS-shielding hydrogel for the delivery of adherent and non-adherent therapeutic cell types. Tissue Engineering Part A. 2017 Oct;23(19-20):1120-1131. doi: 10.1089/ten.tea.2016.0495.
  2. JR Martin, MK Gupta, JM Page, F Yu, JM Davidson, SA Guelcher, CL Duvall . A Porous Tissue Engineering Scaffold Selectively Degraded by Cell-Generated Reactive Oxygen Species. Biomaterials. 2014 Apr; 35(12):3766-76.
  3. MK Gupta, JR Martin, TA Werfel, T Shen, JM Page, CL Duvall. Cell Protective, ABC Triblock Polymer-Based Thermoresponsive Hydrogels with ROS-Triggered Degradation and Drug Release. Journal of the American Chemical Society. 2014, 136 (42), 14896–14902.

Nanomedicine and Targeted Drug Delivery

Development of nanomedicines is a significant focus in the ATL, and this work is broad-reaching in terms of types of materials employed, classes of drugs delivered, and pathological applications pursued.  We develop nanomedicines with two goals in mind: (1) to improve the safety and efficacy of existing drugs and (2) to enable modulation of undruggable targets.  To improve the safety and efficacy of existing drugs, we explore nano-formulation approaches that alter drug pharmacokinetics and increase biodistribution to desired target tissues.  This is achieved through approaches that optimize nanocarrier physical and chemical properties (size, shape, and charge) and through targeting mechanisms (environmental targeting of cues such as enzymes and ROS that are upregulated at pathological sites and more conventional receptor-ligand targeting to promote preferential internalization by specific cell types).  To modulate targets and pathways that are “undruggable” by conventional small molecule drugs, we are developing intracellular acting biologic drugs such as peptide inhibitors, short interfering RNA (siRNA) gene inhibitors, and peptide nucleic acid / locked nucleic acid micro-RNA inhibitors.  These various classes of biologics can enable selective and more robust modulation of protein-protein interactions and certain kinases and enzymes that cannot be robustly and selectively modulated with small molecule approaches.  We utilize polymer based and porous silicon nanocarriers predominantly and pursue applications for systemic delivery nanomedicines including cancer, osteoarthritis, and cardiovascular disease.image004

  1. MJ Uddin, TA Werfel, BC Crews, MK Gupta, TE Kavanaugh, PJ Kingsley, K Boyd, LJ Marnett, CL Duvall. Fluorocoxib A Loaded Nanoparticles Enable Targeted Visualization of Cyclooxygenase-2 in Inflammation and Cancer. Biomaterials. Volume 92, June 2016, Pages 71–80.
  2. TA Werfel, MA Jackson, TE Kavanaugh, KC Kirkbride, M Miteva, TD Giorgio, CL Duvall. Combinatorial Optimization of PEG Architecture and Hydrophobic Content Improves siRNA Polyplex Stability, Pharmacokinetics, and Potency In Vivo. Journal of Controlled Release. 2017 Jun 10;255:12-26. doi: 10.1016/j.jconrel.2017.03.389.
  3. CE Nelson, JR Kintzing, A Hanna, JM Shannon, MK Gupta, CL Duvall. Balancing Cationic and Hydrophobic Content of PEGylated siRNA Polyplexes Enhances Endosome Escape, Stability, Blood Circulation Time, and Bioactivity in Vivo. ACS Nano. 2013, 7(10): 8870-8880.
  4. KR Beavers, TA Werfel, T Shen, TE Kavanaugh, KV Kilchrist, JW Mares, JS Fain, CB Wiese, KC Vickers, SM Weiss, CL Duvall. Porous Silicon and Polymer Nanocomposites for Delivery of Peptide Nucleic Acids as anti-microRNA Therapies. Advanced Materials. 2016 Sep;28(36):7984-7992. doi: 10.1002/adma.201601646.

 


Local, Controlled Release Microparticles and Biomaterials

The Duvall Lab also develops and applies microparticles intended for long-term retention and controlled-release of drug cargo at a local injection site. A recent microparticle system integrates mechanisms for “on demand” drug release modulated by the “need” in the local environment rather than simply prolonging the release, which is enabled by simple hydrolytically-degradable polyester-based constructs.  Specifically, this delivery approach enabled “on demand” release of an antioxidant controlled by the level of reactive oxygen species (ROS) in the local environment.  A recent proof-of-concept application for this microparticle system focused on the delivery of the antioxidant and anti-inflammatory molecule curcumin to promote recovery from ischemic injury in the context of diabetes.  We are pursuing additional applications of this microparticle system in inflammation-associated diseases.  A related research focus (highlighted in a separate section below) is on scaffold-based controlled release of RNAi nanomedicines for sustained, local gene silencing for tissue regenerative applications.

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  1. KM Poole, CE Nelson, RV Joshi, JR Martin, MK Gupta, SC Haws, TE Kavanaugh, MC Skala, CL Duvall. ROS-Responsive Microspheres for On Demand Antioxidant Therapy in a Model of Diabetic Peripheral Arterial Disease. Biomaterials. 2015, (41): 166–175. PMC4274772.
  2. RV Joshi, CE Nelson, KM Poole, MC Skala, CL Duvall. Dual pH- and Temperature-responsive Microparticles for Protein Delivery to Ischemic Tissues. Acta Biomaterialia. 2013 May;9(5):6526-34. doi: 10.1016/j.actbio.2013.01.041.

 


Molecular Design for RNA Albumin Piggybacking

We recently started a new research thrust focused on optimizing albumin “piggybacking” as an elegantly simple natural carrier approach for improving pharmacokinetics of siRNA. RNA molecules have very short circulation time and poor tumor bioavailability due to rapid renal clearance. Designing modified RNAs that dock onto albumin (a long-lasting and highly abundant blood serum protein that is a natural fatty acid carrier) can significantly extend RNA circulation time, tumor accumulation, and homogeneity of tumor penetration. This is a promising strategy for high penetrance delivery of RNAs targeting currently undruggable tumor drivers.

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  1. SM Sarett, TA Werfel, KV Kilchrist, DM Brantley-Sieders, CL Duvall. Lipophilic siRNA Targets Albumin in Situ and Promotes Bioavailability, Tumor Penetration, and Carrier-Free Gene Silencing. PNAS 2017 114 (32) E6490-E6497.
  2. EN Hoogenboezem, CL Duvall. Harnessing Albumin as a Carrier for Cancer Therapies. Advanced Drug Delivery Reviews. 2018 May;130:73-89.

 


Tools for Screening Endosomal Escape and Intracellular Trafficking

One of the major barriers for use of intracellular-acting biologic drugs is the inefficiency of escape from the endo-lysosomal vesicles after cellular entry via endocytosis pathways. There are a lack of quantitative and easily to implement, objective methods for rapidly assessing escape from the endosomal pathway. We are developing tools for both high throughput and in vivo screening of endosomal escape and intracellular bioavailability. In recent work, we validated high throughput methods for in vitro screening for endosome escape based on intracellular redistribution of Galectin 8, a component of the innate immune system, which concentrates onto disrupted endosomes. We are also implementing and developing split GPF and other related technologies for screening delivery formulations ability to disrupt specific endosomal compartments and achieve intracellular bioavailability.

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  1. KV Kilchrist, SC Dimobi, MA Jackson, BC Evans, TA Werfel, EA Dailing, SK Bedingfield, IB Kelly, CL Duvall. Gal8 Visualization of Endosome Disruption Predicts Carrier-mediated Biologic Drug Intracellular Bioavailability. ACS Nano. 2019 Jan 10. doi: 10.1021/acsnano.8b05482. [Epub ahead of print].
  2. KV Kilchrist, BC Evans, CM Brophy, CL Duvall. Mechanism of Enhanced Cellular Uptake and Cytosolic Retention of MK2 Inhibitory Peptide Nano-polyplexes. Cellular and Molecular Bioengineering. 9: 368. doi:10.1007/s12195-016-0446-7. (2016).

 


Pathological Applications


Improved Vascular Graft Function

Another research thrust in the ATL utilizes a pH-responsive, endosomolytic RAFT polymer for intracellular delivery of therapeutic peptides. Current work in this area targets mitogen activated protein kinase (MAPK) signaling pathways that prevent vascular smooth muscle cells (VSMCs) from transitioning to a pathological phenotype. In this application, we seek to inhibit SMC proliferation after vascular graft transplantation, preventing intimal hyperplasia (IH) and ultimately improving long-term graft patency in peripheral and cardiac vascular bypass procedures.
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  1. AJ Mukalel, BC Evans, KV Kilchrist, EA Dailing, J Cheung-Flynn, CM Brophy, CL Duvall. Excipients for the Lyoprotection of MAPKAP Kinase 2 Inhibitory Peptide Nano-Polyplexes. Journal of Controlled Release. Vol 282, July 28, 2018, P. 110-119.
  2. BC Evans, KM Hocking, MJ Osgood, I Voskresensky, J Dmowska, KV Kilchrist, CM Brophy, CL Duvall. Nano-polyplex MK2 Inhibitory Peptide Delivery Inhibits Vascular Graft Intimal Hyperplasia. Science Translational Medicine. 2015 Jun 10;7(291):291ra95.
  3. BC Evans, KM Hocking, KV Kilchrist, ES Wise, CM Brophy, CL Duvall. Endosomolytic Nano-Polyplex Platform Technology for Cytosolic Peptide Delivery to Inhibit Pathological Vasoconstriction. ACS Nano. 2015 Jun 23;9(6):5893-907.

 


Skin Wound Healing

The Duvall Lab also develops polymer-based, controlled delivery systems to efficiently knockdown gene targets to promote tissue formation and repair within nonhealing diabetic skin wounds. We are interested in local modulation of vascularization and also local reprogramming of macrophages to modulate the wound microenvironment to make it more conducive to tissue repair. Our specific focus to this point has been on silencing of PHD2 (prolyl hydroxylase domain 2), which is a negative regulator of the transcription factor HIF1 Silencing PHD2 promotes the expression of numerous pro-angiogenic and pro-healing genes. Other potential applications for this platform technology include repair of bone or critically-sized defects in other tissues.
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  1. JR Martin, CE Nelson, MK Gupta, F Yu, SM Sarett, KM Hocking, AC Pollins, LB Nanney, JM Davidson, SA Guelcher, CL Duvall. Local delivery of PHD2 siRNA from ROS-degradable scaffolds to promote diabetic wound healing. Advanced Healthcare Materials. 2016 Nov;5(21):2751-2757.
  2. CE Nelson, AJ Kim, EJ Adolph, MK Gupta, F Yu, KM Hocking, JM Davidson, SA Guelcher, CL Duvall. Tunable Delivery of siRNA from a Biodegradable Scaffold to Promote Angiogenesis In Vivo. Advanced Materials. 2014 Jan;26(4):607-14.
  3. CE Nelson, MK Gupta, EJ Adolph, JM Shannon, SA Guelcher, CL Duvall. Sustained local delivery of siRNA from an injectable scaffold. Biomaterials. 2012 Feb;33(4):1154-61. PMC3232066.

 


Breast Cancer Therapies

We are currently exploring new breast cancer targets that have not been successfully drugged by small molecule inhibitors, including selective modulation of the different arms of the mTOR signaling pathway using RNA interference. We are also developing and applying targeted nanomedicines that reduce bone destruction as a result of cancer metastasis to bone by drugging key transcription factors.
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  1. TA Werfel, S Wang, MA Jackson, TE Kavanaugh, MM Joly, L Lee, DJ Hicks, VM Sanchez, PI Gonzalez-Ericsson, KV Kilchrist, SC Dimobi, SM Sarett, DM Brantley-Sieders , RS Cook, CL Duvall. Selective mTORC2 inhibitor therapeutically blocks breast cancer cell growth and survival. Cancer Research. 2018 Apr 1;78(7):1845-1858. doi: 10.1158/0008-5472.CAN-17-2388.
  2. MM Williams, L Lee, TA Werfel, MM Joly, DJ Hicks, B Rahman, D Elion, C McKernan, V Sanchez, V Estrada, S Massarweh, R Elledge, CL Duvall, RS Cook. Intrinsic Apoptotic Pathway Activation Increases Response to Anti-Estrogens in Luminal Breast Cancers. Cell Death and Disease. 2018 Jan 17;9(2):21.

 


Osteoarthritis Therapies

Current treatment of osteoarthritis revolves around pain management, and there are not any clinically approved disease modifying osteoarthritis drugs (DMOADs) that actually inhibit or delay this progressive, degenerative, and debilitating disease. We are developing long-lasting microparticle technologies with inherent antioxidant effect and for sustained drug delivery capabilities. We also are having great success in forthcoming work using matrix targeted nanoparticles for sustained retention and selective silencing of OA-driving genes.
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  1. KP O’Grady, TE Kavanaugh, H Cho, H Ye, MK Gupta, MC Madonna, J Lee, CM O’Brien, MC Skala, KA Hasty, CL Duvall. Drug Free ROS Sponge Polymeric Microspheres Reduce Tissue Damage from Ischemic and Mechanical Injury. ACS Biomaterials Science and Engineering. 2018, 4 (4), pp 1251–1264. DOI: 10.1021/acsbiomaterials.6b00804.
  2. TE Kavanaugh, TA Werfel, H Cho, KA Hasty, CL Duvall. Particle-based Technologies for Osteoarthritis Detection and Therapy.  Drug Delivery and Translational Research, 2016 Apr;6(2):132-47. doi: 10.1007/s13346-015-0234-2.