Multivalent Conjugates of Sonic Hedgehog Accelerate Diabetic Wound Healing

Abstract

Despite their preclinical promise, few recombinant growth factors have been fully developed into effective therapies, in part, due to the short interval of therapeutic activity after administration. To address this problem, we developed nanoscale polymer conjugates for multivalent presentation of therapeutic proteins that enhance the activation of targeted cellular responses. As an example of this technology, we conjugated multiple Sonic hedgehog (Shh) proteins onto individual hyaluronic acid biopolymers to generate multivalent protein clusters at defined ratios (i.e., valencies) that yield enhanced Shh pathway activation at equivalent concentrations relative to unconjugated Shh. In this study, we investigated whether these multivalent conjugates (mvShh) could be used to improve the therapeutic function of Shh. We found that a single treatment with mvShh significantly accelerated the closure of full-thickness wounds in diabetic (db/db) mice compared to either an equivalent dose of unconjugated Shh or the vehicle control. Furthermore, we identified specific indicators of wound healing in fibroblasts and endothelial cells (i.e., transcriptional activation and cell migration) that were activated by mvShh in vitro and at concentrations approximately an order of magnitude lower than the unconjugated Shh. Taken together, our findings suggest that mvShh conjugates exhibit greater potency to activate the Shh pathway, and this multivalency advantage improves its therapeutic effect to accelerate wound closure in a diabetic animal model. Our strategy of multivalent protein presentation using nanoscale polymer conjugates has the potential to make a significant impact on the development of protein-based therapies by improving their in vivo performance.

Multivalent hyaluronic acid bioconjugates improve sFlt-1 activity in vitro

“…Conjugation of sFlt to HyA was carried out according to the schematic in Figure 1A, as previously described [16–18]. To make thiol-reactive HyA intermediates, 3,3′- N -(ε-maleimidocaproic acid) hydrazide (EMCH, Thermo Fisher Scientific, 1.2 mg/mL), 1-hydroxybenzotriazole hydrate (HOBt, Sigma Aldrich, 0.3 mg/mL) and 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC, Thermo Fisher Scientific, 10 mg/mL) were added to a 3 mg/mL solution of HyA (Lifecore Biotechnology) of various molecular weights in 0.1 M 2-( N -morpholino)ethanesulphonic acid (MES) (Sigma Aldrich) buffer (pH 6.5) and allowed to react at 4 °C for 4 hours.…”

Glycosaminoglycan and Proteoglycan Biotherapeutics in Articular Cartilage Protection and Repair Strategies: Novel Approaches to Visco‐supplementation in Orthobiologics

“…Multimodal imaging-guided multistage targeted chemo-photothermal therapy [335] Optimized HA-methotrexate conjugates OA therapy [336] Steroids Steroid grafted on to HA Antioxidant delivery [337] EGF HA-EGF conjugate Chronic wound healing [338] Skin wound healing and regeneration [339] Anti-tumor necrosis factor-α Anti-tumor necrosis factor-α HA conjugate Early healing effects in a rat burn model [340] Epigallocatechin-3-O-Gallate HA-Epigallocatechin-3-O-Gallate conjugates injectable hydrogel Free radical scavenging [341] Sonic hedgehog Sonic hedgehog-HA conjugates Wound healing [342] Multivalent Sonic hedgehog conjugate Diabetic wound healing [343] Catechin Hyaluronic acid-green tea catechin nanogels Targeted intracellular protein delivery [344] Quercetin HA-quercetin conjugated micelles A tumor cell-targeted prodrug [345] Opiods HA-opioid conjugate Controlled delivery treatment for pain alleviation applications [346] Adv. Therap.…”

Biomaterials-based strategies for salivary gland tissue regeneration

“…Non-covalent interactions 78 or reversible covalent bonds 79, 80 can be introduced to impart dynamic properties to the matrices without compromising the network integrity. A programmed introduction of various biological signals, in a multivalent fashion, 81, 82 can promote the desired cellular functions at different stages of tissue assembly. 83 …”