Controlled release of MIF siRNA and GDNF protein from a photocurable scaffold efficiently repairs spinal cord injury

Abstract

Compared with traditional treatment strategies, siRNA-based gene therapy combines with protein therapy to offer a new strategy for spinal cord injury (SCI). The siRNA and protein therapy are limited by the large and deep lesion site and local co-delivery vectors. However, the photocurable scaffold has the properties of injectable, flexible, and biodegradable, which provide a potential formulation for siRNA and protein combined therapy. Here, a photocurable lipid nanoparticle gel (PLNG) scaffold is designed for efficiently sustained and controlled release of the macrophage migration-inhibitory factor (MIF) targeted siRNA and co-delivery of GDNF protein for SCI. The GDNF is chemically modified in the scaffold and the prepared GDNF-PLNG/siRNA scaffold is injectable with easily photocured. This formulation can inhibit inflammation by promoting macrophage M2 polarization and effectively promote primary neuron axon growth. After locally administered with GDNF-PLNG/siMIF scaffold to SCI mice, the scaffold promoted neuron regeneration by upregulation of neuron cytokine production and inhibited inflammation through the downregulation immune pathway. With the interaction mechanism of GDNF and MIF siRNA, GDNF-PLNG/siMIF scaffold increases the collagen and integrin expression to promote spinal cord repairing and significantly improve motor function, so that scaffold is a potential candidate gene formulation applied to clinical SCI treatment.

Keywords

controlled-release / hydrogel / nanoparticle / siRNA-based gene therapy / spinal cord injury

References

[1] Krause JS, Cao Y, DiPiro N. Psychological factors and risk of mortality after spinal cord injury. J Spinal Cord Med. 2020; 43(5): 667-675.

[2] Griffin JM, Bradke F. Therapeutic repair for spinal cord injury: combinatory approaches to address a multifaceted problem. EMBO Mol Med. 2020; 12(3): e11505.

[3] Gao Y, Men K, Pan C, et al. Functionalized DMP-039 hybrid nanoparticle as a novel mRNA vector for efficient cancer suicide gene therapy. Int J Nanomedicine. 2021; 16: 5211-5232.

[4] Lei S, Zhang X, Men K, et al. Efficient colorectal cancer gene therapy with IL-15 mRNA nanoformulation. Molecular pharmaceutics. 2020; 17(9): 3378-3391.

[5] Lei S, Chen X, Gao Y, et al. ALPPL2-binding peptide facilitates targeted mrna delivery for efficient hepatocellular carcinoma gene therapy. Advanced Functional Materials. 2022:2204342.

[6] Yang X, Yang W, Xia X, et al. Intranasal delivery of BACE1 siRNA and rapamycin by dual targets modified nanoparticles for Alzheimer’s disease therapy. Small. 2022; 18(30): e2203182.

[7] Kandil R, Baldassi D, Bohlen S, et al. Targeted GATA3 knockdown in activated T cells via pulmonary siRNA delivery as novel therapy for allergic asthma. J Control Release. 2023; 354: 305-315.

[8] Shan S, Chen J, Sun Y, et al. Functionalized macrophage exosomes with panobinostat and PPM1D-siRNA for diffuse intrinsic pontine gliomas therapy. Adv Sci (Weinh). 2022; 9(21): e2200353.

[9] Guo S, Perets N, Betzer O, et al. Intranasal delivery of mesenchymal stem cell derived exosomes loaded with phosphatase and tensin homolog sirna repairs complete spinal cord injury. ACS Nano. 2019; 13(9): 10015-10028.

[10] Rong Y, Wang Z, Tang P, et al. Engineered extracellular vesicles for delivery of siRNA promoting targeted repair of traumatic spinal cord injury. Bioact Mater. 2023; 23: 328-342.

[11] Francos-Quijorna I, Sanchez-Petidier M, Burnside ER, et al. Chondroitin sulfate proteoglycans prevent immune cell phenotypic conversion and inflammation resolution via TLR4 in rodent models of spinal cord injury. Nat Commun. 2022; 13(1): 2933.

[12] Lee D, Baek S, Kim YY, et al. Self-assembled DNA-protein hybrid nanospheres: biocompatible nano-drug-carriers for targeted cancer therapy. ACS Appl Mater Interfaces. 2022; 14(33): 37493-37503.

[13] De Lorenzo F, Luningschror P, Nam J, et al. CDNF rescues motor neurons in models of amyotrophic lateral sclerosis by targeting endoplasmic reticulum stress. Brain. 2023; 146(9): 3783-3799.

[14] Qian K, Bao X, Li Y, et al. Cholinergic neuron targeting nanosystem delivering hybrid peptide for combinatorial mitochondrial therapy in Alzheimer’s disease. ACS Nano. 2022; 16(7): 11455-11472.

[15] Ma J, Li J, Wang X, et al. GDNF-loaded polydopamine nanoparticles-based anisotropic scaffolds promote spinal cord repair by modulating inhibitory microenvironment. Adv Healthcare Mater. 2023; 12(8): e2202377.

[16] Wang L, Zhang D, Ren Y, et al. Injectable hyaluronic acid hydrogel loaded with BMSC and NGF for traumatic brain injury treatment. Mater Today Bio. 2022; 13: 100201.

[17] Zhang F, Wu X, Li Q, et al. Dual growth factor methacrylic alginate microgels combined with chitosan-based conduits facilitate peripheral nerve repair. Int J Biol Macromol. 2024; 268(1): 131594. Pt.

[18] Shen H, Xu B, Yang C, et al. A DAMP-scavenging, IL-10-releasing hydrogel promotes neural regeneration and motor function recovery after spinal cord injury. Biomaterials. 2022; 280: 121279.

[19] Jaffer H, Andrabi SS, Petro M, Kuang Y, Steinmetz MP, Labhasetwar V. Catalytic antioxidant nanoparticles mitigate secondary injury progression and promote functional recovery in spinal cord injury model. J Control Release. 2023; 364: 109-123.

[20] Feng F, Song X, Tan Z, et al. Cooperative assembly of a designer peptide and silk fibroin into hybrid nanofiber gels for neural regeneration after spinal cord injury. Sci Adv. 2023; 9(25): eadg0234.

[21] Li J, Yao Y, Wang Y, et al. Modulation of the crosstalk between Schwann cells and macrophages for nerve regeneration: a therapeutic strategy based on a multifunctional tetrahedral framework nucleic acids system. Adv Mater. 2022; 34(46): e2202513.

[22] Chen P, Xu C, Wu P, et al. Wirelessly powered electrical-stimulation based on biodegradable 3D piezoelectric scaffolds promotes the spinal cord injury repair. ACS Nano. 2022; 16(10): 16513-16528.

[23] Xin W, Baokun Z, Zhiheng C, et al. Biodegradable bilayer hydrogel membranes loaded with bazedoxifene attenuate blood-spinal cord barrier disruption via the NF-kappaB pathway after acute spinal cord injury. Acta Biomater. 2023; 159: 140-155.

[24] Zeng X, Wei QS, Ye JC, et al. A biocompatible gelatin sponge scaffold confers robust tissue remodeling after spinal cord injury in a non-human primate model. Biomaterials. 2023; 299: 122161.

[25] Sha Q, Wang Y, Zhu Z, et al. A hyaluronic acid/silk fibroin/poly-dopamine-coated biomimetic hydrogel scaffold with incorporated neurotrophin-3 for spinal cord injury repair. Acta Biomater. 2023; 167: 219-233.

[26] Gao C, Li Y, Liu X, Huang J, Zhang Z. 3D bioprinted conductive spinal cord biomimetic scaffolds for promoting neuronal differentiation of neural stem cells and repairing of spinal cord injury. Chem Eng J. 2023; 451: 138788.

[27] Wang R, Wu X, Tian Z, et al. Sustained release of hydrogen sulfide from anisotropic ferrofluid hydrogel for the repair of spinal cord injury. Bioact Mater. 2023; 23: 118-128.

[28] Silva D, Schirmer L, Pinho TS, et al. Sustained release of human adipose tissue stem cell secretome from star-shaped poly(ethylene glycol) glycosaminoglycan hydrogels promotes motor improvements after complete transection in spinal cord injury rat model. Adv Healthcare Mater. 2023; 12(17): e2202803.

[29] Liu X, Song S, Chen Z, et al. Release of O-GlcNAc transferase inhibitor promotes neuronal differentiation of neural stem cells in 3D bioprinted supramolecular hydrogel scaffold for spinal cord injury repair. Acta Biomater. 2022; 151: 148-162.

[30] Gao Y, Wang K, Wu S, et al. Injectable and photocurable gene scaffold facilitates efficient repair of spinal cord injury. ACS Appl Mater Interface. 2024; 16(4): 4375-4394.

[31] Zhang J, Li Y, Xiong J, et al. Delivery of pOXR1 through an injectable liposomal nanoparticle enhances spinal cord injury regeneration by alleviating oxidative stress. Bioact Mater. 2021; 6(10): 3177-3191.

[32] Song Z, Wang Z, Shen J, Xu S, Hu Z, Nerve growth factor delivery by ultrasound-mediated nanobubble destruction as a treatment for acute spinal cord injury in rats. Int J Nanomedicine. 2017; 12: 1717-1729.

[33] Leibinger M, Zeitler C, Gobrecht P, Andreadaki A, Gisselmann G, Fischer D. Transneuronal delivery of hyper-interleukin-6 enables functional recovery after severe spinal cord injury in mice. Nat Commun. 2021; 12(1): 391.

[34] Bavencoffe AG, Spence EA, Zhu MY, et al. Macrophage migration inhibitory factor (MIF) makes complex contributions to pain-related hyperactivity of nociceptors after spinal cord injury. J Neurosci. 2022; 42(27): 5463-5480.

[35] Zhang H, Hu YM, Wang YJ, et al. Macrophage migration inhibitory factor facilitates astrocytic production of the CCL2 chemokine following spinal cord injury. Neural Regen Res. 2023; 18(8): 1802-1808.

[36] Piri SM, Ghodsi Z, Shool S, et al. Macrophage migration inhibitory factor as a therapeutic target after traumatic spinal cord injury: a systematic review. Eur Spine J. 2021; 30(6): 1474-1494.

[37] Zhang Y, Zhou Y, Chen S, et al. Macrophage migration inhibitory factor facilitates prostaglandin E2 production of astrocytes to tune inflammatory milieu following spinal cord injury. J Neuroinflammation. 2019; 16(1): 85.

[38] Zhou Y, Guo W, Zhu Z, et al. Macrophage migration inhibitory factor facilitates production of CCL5 in astrocytes following rat spinal cord injury. J Neuroinflammation. 2018; 15(1): 253.

[39] Baloh RH, Johnson JP, Avalos P, et al. Transplantation of human neural progenitor cells secreting GDNF into the spinal cord of patients with ALS: a phase 1/2a trial. Nat Med. 2022; 28(9): 1813-1822.

[40] Hunt CPJ, Penna V, Gantner CW, et al. Tissue programmed hydrogels functionalized with GDNF improve human neural grafts in Parkinson’s disease. Adv Funct Mater. 2021; 31(47)

[41] Zheng D, Wang K, Bai B, Hu N, Wang H. Swelling and glyphosate-controlled release behavior of multi-responsive alginate-g-P(NIPAm-co-NDEAm)-based hydrogel. Carbohydr Polym. 2022; 282: 119113.

[42] Teal CJ, Hettiaratchi MH, Ho MT, et al. Directed evolution enables simultaneous controlled release of multiple therapeutic proteins from biopolymer-based hydrogels. Adv Mater. 2022; 34(34): e2202612.

[43] Phan VHG, Murugesan M, Huong H, et al. Cellulose nanocrystals-incorporated thermosensitive hydrogel for controlled release, 3D printing, and breast cancer treatment applications. ACS Appl Mater Interfaces. 2022; 14(38): 42812-42826.

[44] Nazemi Z, Nourbakhsh MS, Kiani S, et al. Co-delivery of minocycline and paclitaxel from injectable hydrogel for treatment of spinal cord injury. J Control Release. 2020; 321: 145-158.

[45] Boido M, Ghibaudi M, Gentile P, Favaro E, Fusaro R, Tonda-Turo C. Chitosan-based hydrogel to support the paracrine activity of mesenchymal stem cells in spinal cord injury treatment. Sci Rep. 2019; 9(1): 6402.

[46] Zhao YZ, Jiang X, Lin Q, et al. Thermosensitive heparin-poloxamer hydrogels enhance the effects of GDNF on neuronal circuit remodeling and neuroprotection after spinal cord injury. J Biomed Mater Res A. 2017; 105(10): 2816-2829.

[47] Long Y, Yan L, Dai H, et al. Enhanced proliferation and differentiation of neural stem cells by peptide-containing temperature-sensitive hydrogel scaffold. Mater Sci Eng C Mater Biol Appl. 2020; 116: 111258.

[48] Han Z, Wang P, Mao G, et al. Dual pH-responsive hydrogel actuator for lipophilic drug delivery. ACS Appl Mater Interfaces. 2020; 12(10): 12010-12017.

[49] Nguyen HT, Massino M, Keita C, Salmon JB. Microfluidic dialysis using photo-patterned hydrogel membranes in PDMS chips. Lab Chip. 2020; 20(13): 2383-2393.

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