Orally Delivered Connexin43 Hemichannel Blocker, Tonabersat, Inhibits Vascular Breakdown and Inflammasome Activation in a Mouse Model of Diabetic Retinopathy
Abstract
:1. Introduction
2. Results
2.1. Tonabersat Did Not Induce Any Changes in Uninjured ARPE-19 Cells or NOD Mice
2.2. Tonabersat Decreased the Incidence of Macrovascular Abnormalities
2.3. Tonabersat Prevented Retinal Hyperreflective Foci Formation, Swelling, and Subretinal Fluid Accumulation
2.4. Tonabersat Decreased the Number of PLVAP+ Vessels and the Number of Connexin43 Spots on PLVAP+ Vessels
2.5. Tonabersat Inhibited GFAP Upregulation and Iba-1+ Cell ONL Infiltration
2.6. Tonabersat Inhibited NLRP3 and Cleaved Caspase-1 Upregulation
3. Discussion
4. Materials and Methods
4.1. Animal Studies
4.2. Intravitreal Injection of Pro-Inflammatory Cytokines
4.3. Tonabersat Treatment
4.4. Funduscopy and Optical Coherence Tomography (OCT) Imaging
4.5. Immunohistochemistry
4.6. Statistical Analysis
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
- Abderrazak, A.; Syrovets, T.; Couchie, D.; El Hadri, K.; Friguet, B.; Simmet, T.; Rouis, M. NLRP3 inflammasome: From a danger signal sensor to a regulatory node of oxidative stress and inflammatory diseases. Redox Biol. 2015, 4, 296–307. [Google Scholar] [CrossRef]
- Leemans, J.C.; Cassel, S.L.; Sutterwala, F.S. Sensing damage by the NLRP3 inflammasome. Immunol. Rev. 2011, 243, 152–162. [Google Scholar] [CrossRef] [PubMed]
- Wang, Z.; Zhang, S.; Xiao, Y.; Zhang, W.; Wu, S.; Qin, T.; Yue, Y.; Qian, W.; Li, L. NLRP3 Inflammasome and Inflammatory Diseases. Oxid. Med. Cell. Longev. 2020, 2020, 4063562. [Google Scholar] [CrossRef] [PubMed]
- Chaurasia, S.S.; Lim, R.R.; Parikh, B.H.; Wey, Y.S.; Tun, B.B.; Wong, T.Y.; Luu, C.D.; Agrawal, R.; Ghosh, A.; Mortellaro, A. The NLRP3 inflammasome may contribute to pathologic neovascularization in the advanced stages of diabetic retinopathy. Sci. Rep. 2018, 8, 2847. [Google Scholar] [CrossRef] [PubMed]
- Chen, H.; Zhang, X.; Liao, N.; Mi, L.; Peng, Y.; Liu, B.; Zhang, S.; Wen, F. Enhanced expression of NLRP3 inflammasome-related inflammation in diabetic retinopathy. Investig. Ophthalmol. Vis. Sci. 2018, 59, 978–985. [Google Scholar] [CrossRef]
- Loukovaara, S.; Piippo, N.; Kinnunen, K.; Hytti, M.; Kaarniranta, K.; Kauppinen, A. NLRP3 inflammasome activation is associated with proliferative diabetic retinopathy. Acta Ophthalmol. 2017, 95, 803–808. [Google Scholar] [CrossRef]
- Kuo, C.Y.J.; Murphy, R.; Rupenthal, I.D.; Mugisho, O.O. Correlation between the progression of diabetic retinopathy and inflammasome biomarkers in vitreous and serum—A systematic review. BMC Ophthalmol. 2022, 22, 238. [Google Scholar] [CrossRef]
- Acosta, M.L.; Mat Nor, M.N.; Guo, C.X.; Mugisho, O.O.; Coutinho, F.P.; Rupenthal, I.D.; Green, C.R. Connexin therapeutics: Blocking connexin hemichannel pores is distinct from blocking pannexin channels or gap junctions. Neural Regen. Res. 2021, 16, 482–488. [Google Scholar] [CrossRef]
- Mat Nor, M.N.; Rupenthal, I.D.; Green, C.R.; Acosta, M.L. Differential Action of Connexin Hemichannel and Pannexin Channel Therapeutics for Potential Treatment of Retinal Diseases. Int. J. Mol. Sci. 2021, 22, 1755. [Google Scholar] [CrossRef]
- Mugisho, O.O.; Green, C.R.; Kho, D.T.; Zhang, J.; Graham, E.S.; Acosta, M.L.; Rupenthal, I.D. The inflammasome pathway is amplified and perpetuated in an autocrine manner through connexin43 hemichannel mediated ATP release. Biochim. Biophys. Acta 2018, 1862, 385–393. [Google Scholar] [CrossRef]
- Mugisho, O.O.; Green, C.R.; Squirrell, D.M.; Bould, S.; Danesh-Meyer, H.V.; Zhang, J.; Acosta, M.L.; Rupenthal, I.D. Connexin43 hemichannel block protects against the development of diabetic retinopathy signs in a mouse model of the disease. J. Mol. Med. 2019, 97, 215–229. [Google Scholar] [CrossRef] [PubMed]
- Mugisho, O.O.; Rupenthal, I.D.; Paquet-Durand, F.; Acosta, M.L.; Green, C.R. Targeting connexin hemichannels to control the inflammasome: The correlation between connexin43 and NLRP3 expression in chronic eye disease. Expert Opin. Ther. Targets 2019, 23, 855–863. [Google Scholar] [CrossRef] [PubMed]
- Mugisho, O.O.; Rupenthal, I.D.; Squirrell, D.M.; Bould, S.J.; Danesh-Meyer, H.V.; Zhang, J.; Green, C.R.; Acosta, M.L. Intravitreal pro-inflammatory cytokines in non-obese diabetic mice: Modelling signs of diabetic retinopathy. PLoS ONE 2018, 13, e0202156. [Google Scholar] [CrossRef] [PubMed]
- Mugisho, O.O.; Green, C.R.; Squirrell, D.; Bould, S.; Zhang, J.; Acosta, M.; Rupenthal, I.D. Intravitreal pro-inflammatory cytokines induce signs of diabetic retinopathy in non-obese diabetic mice. Investig. Ophthalmol. Vis. Sci. 2018, 59, 5358. [Google Scholar]
- Louie, H.H.; Shome, A.; Kuo, C.Y.J.; Rupenthal, I.D.; Green, C.R.; Mugisho, O.O. Connexin43 hemichannel block inhibits NLRP3 inflammasome activation in a human retinal explant model of diabetic retinopathy. Exp. Eye Res. 2021, 202, 108384. [Google Scholar] [CrossRef]
- Lyon, H.; Shome, A.; Rupenthal, I.D.; Green, C.R.; Mugisho, O.O. Tonabersat Inhibits Connexin43 Hemichannel Opening and Inflammasome Activation in an In Vitro Retinal Epithelial Cell Model of Diabetic Retinopathy. Int. J. Mol. Sci. 2020, 22, 298. [Google Scholar] [CrossRef]
- Bialer, M.; Johannessen, S.I.; Levy, R.H.; Perucca, E.; Tomson, T.; White, H.S. Progress report on new antiepileptic drugs: A summary of the Ninth Eilat Conference (EILAT IX). Epilepsy Res. 2009, 83, 1–43. [Google Scholar] [CrossRef]
- Abcouwer, S.F.; Gardner, T.W. Diabetic retinopathy: Loss of neuroretinal adaptation to the diabetic metabolic environment. Ann. N. Y. Acad. Sci. 2014, 1311, 174. [Google Scholar] [CrossRef]
- Comparison of Age-Related Macular Degeneration Treatments Trials (CATT) Research Group; Martin, D.F.; Maguire, M.G.; Fine, S.L.; Ying, G.-S.; Jaffe, G.J.; Grunwald, J.E.; Toth, C.; Redford, M.; Ferris, F.L., 3rd. Ranibizumab and bevacizumab for treatment of neovascular age-related macular degeneration: Two-year results. Ophthalmology 2012, 119, 1388–1398. [Google Scholar] [CrossRef]
- Whitehead, M.; Wickremasinghe, S.; Osborne, A.; Van Wijngaarden, P.; Martin, K.R. Diabetic retinopathy: A complex pathophysiology requiring novel therapeutic strategies. Expert Opin. Biol. Ther. 2018, 18, 1257–1270. [Google Scholar] [CrossRef]
- Wong, T.Y.; Sun, J.; Kawasaki, R.; Ruamviboonsuk, P.; Gupta, N.; Lansingh, V.C.; Maia, M.; Mathenge, W.; Moreker, S.; Muqit, M.M. Guidelines on diabetic eye care: The international council of ophthalmology recommendations for screening, follow-up, referral, and treatment based on resource settings. Ophthalmology 2018, 125, 1608–1622. [Google Scholar] [CrossRef] [PubMed]
- Stewart, M.W.; Flynn, H.W., Jr.; Schwartz, S.G.; Scott, I.U. Extended duration strategies for the pharmacologic treatment of diabetic retinopathy: Current status and future prospects. Expert Opin. Drug Deliv. 2016, 13, 1277–1287. [Google Scholar] [CrossRef] [PubMed]
- Kim, Y.; Griffin, J.M.; Nor, M.N.M.; Zhang, J.; Freestone, P.S.; Danesh-Meyer, H.V.; Rupenthal, I.D.; Acosta, M.; Nicholson, L.F.B.; O’Carroll, S.J.; et al. Tonabersat Prevents Inflammatory Damage in the Central Nervous System by Blocking Connexin43 Hemichannels. Neurotherapeutics 2017, 14, 1148–1165. [Google Scholar] [CrossRef] [PubMed]
- Dahlof, C.G.; Hauge, A.W.; Olesen, J. Efficacy and safety of tonabersat, a gap-junction modulator, in the acute treatment of migraine: A double-blind, parallel-group, randomized study. Cephalalgia 2009, 29 (Suppl. S2), 7–16. [Google Scholar] [CrossRef]
- Coscas, G.; Lupidi, M.; Coscas, F. OCT-A: Guided treatment of diabetic retinopathy. Acta Ophthalmol. 2017, 95, S259. [Google Scholar] [CrossRef]
- Ajlan, R.S.; Silva, P.S.; Sun, J.K. Vascular Endothelial Growth Factor and Diabetic Retinal Disease; Taylor & Francis: Abingdon, UK, 2016; pp. 40–48. [Google Scholar]
- Antonetti, D.A.; Barber, A.J.; Bronson, S.K.; Freeman, W.M.; Gardner, T.W.; Jefferson, L.S.; Kester, M.; Kimball, S.R.; Krady, J.K.; LaNoue, K.F.; et al. Diabetic retinopathy: Seeing beyond glucose-induced microvascular disease. Diabetes 2006, 55, 2401–2411. [Google Scholar] [CrossRef]
- Antonetti, D.A.; Lieth, E.; Barber, A.J.; Gardner, T.W. Molecular mechanisms of vascular permeability in diabetic retinopathy. Semin. Ophthalmol. 1999, 14, 240–248. [Google Scholar] [CrossRef]
- Niemela, H.; Elima, K.; Henttinen, T.; Irjala, H.; Salmi, M.; Jalkanen, S. Molecular identification of PAL-E, a widely used endothelial-cell marker. Blood 2005, 106, 3405–3409. [Google Scholar] [CrossRef]
- Wisniewska-Kruk, J.; van der Wijk, A.-E.; van Veen, H.A.; Gorgels, T.G.M.F.; Vogels, I.M.C.; Versteeg, D.; Van Noorden, C.J.F.; Schlingemann, R.O.; Klaassen, I. Plasmalemma Vesicle-Associated Protein Has a Key Role in Blood-Retinal Barrier Loss. Am. J. Pathol. 2016, 186, 1044–1054. [Google Scholar] [CrossRef]
- Schlingemann, R.O.; Hofman, P.; Andersson, L.; Troost, D.; van der Gaag, R. Vascular Expression of Endothelial Antigen PAL-E Indicates Absence of Blood-Ocular Barriers in the Normal Eye. Ophthalmic. Res. 1997, 29, 130–138. [Google Scholar] [CrossRef]
- Shue, E.H.; Carson-Walter, E.B.; Liu, Y.; Winans, B.N.; Ali, Z.S.; Chen, J.; Walter, K.A. Plasmalemmal Vesicle Associated Protein-1 (PV-1) is a marker of blood-brain barrier disruption in rodent models. BMC Neurosci. 2008, 9, 29. [Google Scholar] [CrossRef] [PubMed]
- Schlingemann, R.O.; Hofman, P.; Vrensen, G.F.J.M.; Blaauwgeers, H.G.T. Increased expression of endothelial antigen PAL-E in human diabetic retinopathy correlates with microvascular leakage. Diabetologia 1999, 42, 596–602. [Google Scholar] [CrossRef] [PubMed]
- Wisniewska-Kruk, J.; Klaassen, I.; Vogels, I.M.; Van Noorden, C.J.; Schlingemann, R.O.; Group, O.A. PLVAP Modulates Angiogenesis By Tuning VEGF Signaling In Endothelial Cells. Investig. Ophthalmol. Vis. Sci. 2014, 55, 2241. [Google Scholar]
- Wisniewska-Kruk, J.; Klaassen, I.; Vogels, I.M.; Magno, A.L.; Lai, C.M.; Van Noorden, C.J.; Schlingemann, R.O.; Rakoczy, E.P. Molecular analysis of blood-retinal barrier loss in the Akimba mouse, a model of advanced diabetic retinopathy. Exp. Eye Res. 2014, 122, 123–131. [Google Scholar] [CrossRef]
- Guo, L.; Zhang, H.; Hou, Y.; Wei, T.; Liu, J. Plasmalemma vesicle-associated protein: A crucial component of vascular homeostasis. Exp. Ther. Med. 2016, 12, 1639–1644. [Google Scholar] [CrossRef] [PubMed]
- Williams, M.D.; Nadler, J.L. Inflammatory mechanisms of diabetic complications. Curr. Diab. Rep. 2007, 7, 242–248. [Google Scholar] [CrossRef]
- Youngblood, H.; Robinson, R.; Sharma, A.; Sharma, S. Proteomic biomarkers of retinal inflammation in diabetic retinopathy. Int. J. Mol. Sci. 2019, 20, 4755. [Google Scholar] [CrossRef]
- Zhang, W.; Liu, H.; Rojas, M.; Caldwell, R.W.; Caldwell, R.B. Anti-inflammatory therapy for diabetic retinopathy. Immunotherapy 2011, 3, 609–628. [Google Scholar] [CrossRef]
- Das, U.N. Diabetic macular edema, retinopathy and age-related macular degeneration as inflammatory conditions. Arch. Med. Sci. 2016, 12, 1142–1157. [Google Scholar] [CrossRef]
- Tittarelli, A. Connexin channels modulation in pathophysiology and treatment of immune and inflammatory disorders. Biochim. Biophys. Acta Mol. Basis Dis. 2021, 1867, 166258. [Google Scholar] [CrossRef]
- Acosta, M.L.; Mat Nor, N.; Guo, C.X.; Rupenthal, I.D.; Green, C.R. A hemichannel blocker currently undergoing clinical trials reduces inflammation and prevents retinal damage in an animal model of age related macular degeneration. In Proceedings of the International Symposium on Ocular Pharmacology and Therapeutics, Rome, Italy, 2 December 2016. [Google Scholar]
- Nor, M.N.M.; Rupenthal, I.D.; Green, C.R.; Acosta, M.L. Connexin Hemichannel Block Using Orally Delivered Tonabersat Improves Outcomes in Animal Models of Retinal Disease. Neurotherapeutics 2020, 17, 371–387. [Google Scholar]
- Vujosevic, S.; Micera, A.; Bini, S.; Berton, M.; Esposito, G.; Midena, E. Aqueous Humor Biomarkers of Müller Cell Activation in Diabetic Eyes. Investig. Ophthalmol. Vis. Sci. 2015, 56, 3913–3918. [Google Scholar] [CrossRef]
- Curtis, T.M.; Hamilton, R.; Yong, P.H.; McVicar, C.M.; Berner, A.; Pringle, R.; Uchida, K.; Nagai, R.; Brockbank, S.; Stitt, A.W. Müller glial dysfunction during diabetic retinopathy in rats is linked to accumulation of advanced glycation end-products and advanced lipoxidation end-products. Diabetologia 2011, 54, 690–698. [Google Scholar] [CrossRef] [PubMed]
- Wang, J.; Xu, X.; Elliott, M.H.; Zhu, M.; Le, Y.-Z. Müller Cell-Derived VEGF Is Essential for Diabetes-Induced Retinal Inflammation and Vascular Leakage. Diabetes 2010, 59, 2297–2305. [Google Scholar] [CrossRef] [PubMed]
- Abcouwer, S.F. Müller Cell–Microglia Cross Talk Drives Neuroinflammation in Diabetic Retinopathy. Diabetes 2017, 66, 261–263. [Google Scholar] [CrossRef] [PubMed]
- Portillo, J.-A.C.; Lopez Corcino, Y.; Miao, Y.; Tang, J.; Sheibani, N.; Kern, T.S.; Dubyak, G.R.; Subauste, C.S. CD40 in Retinal Müller Cells Induces P2X7-Dependent Cytokine Expression in Macrophages/Microglia in Diabetic Mice and Development of Early Experimental Diabetic Retinopathy. Diabetes 2016, 66, 483–493. [Google Scholar] [CrossRef] [PubMed]
- Barathi, V.A.; Lim, R.R.; Parikh, B.H.; Wey, Y.S.; Wong, T.Y.; Mortellaro, A.; Chaurasia, S.S. Activation of NLRP3 inflammasome in proliferative diabetic retinopathy. Investig. Ophthalmol. Vis. Sci. 2015, 56, 3487. [Google Scholar]
- Chen, W.; Zhao, M.; Zhao, S.; Lu, Q.; Ni, L.; Zou, C.; Lu, L.; Xu, X.; Guan, H.; Zheng, Z. Activation of the TXNIP/NLRP3 inflammasome pathway contributes to inflammation in diabetic retinopathy: A novel inhibitory effect of minocycline. Inflamm. Res. 2017, 66, 157–166. [Google Scholar] [CrossRef]
- Harkin, K.; Chen, M.; Xu, H.; Pavlou, S. The inflammasome pathway is activated in the retina of type 2 but not type 1 diabetic mice. Investig. Ophthalmol. Vis. Sci. Conf. 2019, 60, 768. [Google Scholar]
- Raman, K.S.; Matsubara, J.A. Dysregulation of the NLRP3 Inflammasome in Diabetic Retinopathy and Potential Therapeutic Targets. Ocul. Immunol. Inflamm. 2020, 30, 470–478. [Google Scholar] [CrossRef]
- Alyaseer, A.A.A.; de Lima, M.H.S.; Braga, T.T. The Role of NLRP3 Inflammasome Activation in the Epithelial to Mesenchymal Transition Process During the Fibrosis. Front. Immunol. 2020, 11, 883. [Google Scholar] [CrossRef] [PubMed]
- Baroja-Mazo, A.; Martin-Sanchez, F.; Gomez, A.I.; Martinez, C.M.; Amores-Iniesta, J.; Compan, V.; Barbera-Cremades, M.; Yague, J.; Ruiz-Ortiz, E.; Anton, J.; et al. The NLRP3 inflammasome is released as a particulate danger signal that amplifies the inflammatory response. Nat. Immunol. 2014, 15, 738–748. [Google Scholar] [CrossRef] [PubMed]
- Butts, B.; Gary, R.A.; Dunbar, S.B.; Butler, J. The Importance of NLRP3 Inflammasome in Heart Failure. J. Card. Fail. 2015, 21, 586–593. [Google Scholar] [CrossRef] [PubMed]
- Celkova, L.; Doyle, S.L.; Campbell, M. NLRP3 Inflammasome and Pathobiology in AMD. J. Clin. Med. 2015, 4, 172–192. [Google Scholar] [CrossRef] [PubMed]
- Coll, R.C.; Robertson, A.A.B.; Chae, J.J.; Higgins, S.C.; Munoz-Planillo, R.; Inserra, M.C.; Vetter, I.; Dungan, L.S.; Monks, B.G.; Stutz, A.; et al. A small-molecule inhibitor of the NLRP3 inflammasome for the treatment of inflammatory diseases. Nat. Med. 2015, 21, 248–255. [Google Scholar] [CrossRef]
- de Rivero Vaccari, J.P.; Dietrich, W.D.; Keane, R.W. Activation and regulation of cellular inflammasomes: Gaps in our knowledge for central nervous system injury. J. Cereb. Blood Flow Metab. 2014, 34, 369–375. [Google Scholar] [CrossRef]
- Dupont, M.D.; Longhini, A.L.; Sreejit, G.; Prasad, R.; Nagareddy, P.; Grant, M.B. Circulating inflammatory cells and inflammasome activation in diabetics with diabetic retinopathy (DR). Diabetes 2019, 68 (Suppl. S1), 603. [Google Scholar] [CrossRef]
- Gao, J.; Liu, R.T.; Cao, S.; Cui, J.Z.; Wang, A.; To, E.; Matsubara, J.A. NLRP3 inflammasome: Activation and regulation in age-related macular degeneration. Mediat. Inflamm. 2015, 2015, 690243. [Google Scholar] [CrossRef]
- Gao, L.; Dong, Q.; Song, Z.; Shen, F.; Shi, J.; Li, Y. NLRP3 inflammasome: A promising target in ischemic stroke. Inflamm. Res. 2017, 66, 17–24. [Google Scholar] [CrossRef]
- Hosseinian, N.; Cho, Y.; Lockey, R.F.; Kolliputi, N. The role of the NLRP3 inflammasome in pulmonary diseases. Ther. Adv. Respir. Dis. 2015, 9, 188–197. [Google Scholar] [CrossRef]
- Ildefonso, C.J.; Biswal, M.R.; Ahmed, C.M.; Lewin, A.S. The NLRP3 Inflammasome and its Role in Age-Related Macular Degeneration. In Retinal Degenerative Diseases; Springer: Berlin/Heidelberg, Germany, 2016; pp. 59–65. [Google Scholar]
- Kerur, N.; Hirano, Y.; Tarallo, V.; Fowler, B.J.; Bastos-Carvalho, A.; Yasuma, T.; Yasuma, R.; Kim, Y.; Hinton, D.R.; Kirschning, C.J.; et al. TLR-Independent and P2X7-Dependent Signaling Mediate Alu RNA-Induced NLRP3 Inflammasome Activation in Geographic Atrophy. Investig. Ophthalmol. Vis. Sci. 2013, 54, 7395–7401. [Google Scholar] [CrossRef] [PubMed]
- Liao, Y.; Zhang, H.J.; He, D.X.; Wang, Y.; Cai, B.X.; Chen, J.M.; Ma, J.X.; Liu, Z.G.; Wu, Y.L. Retinal Pigment Epithelium Cell Death Is Associated With NLRP3 Inflammasome Activation by All-trans Retinal. Investig. Ophthalmol. Vis. Sci. 2019, 60, 3034–3045. [Google Scholar] [CrossRef] [PubMed]
- Liberale, L.; Montecucco, F.; Tardif, J.C.; Libby, P.; Camici, G.G. Inflamm-ageing: The role of inflammation in age-dependent cardiovascular disease. Eur. Heart J. 2020, 41, 2974–2982. [Google Scholar] [CrossRef] [PubMed]
- Marín-Aguilar, F.; Lechuga-Vieco, A.V.; Alcocer-Gómez, E.; Castejón-Vega, B.; Lucas, J.; Garrido, C.; Peralta-Garcia, A.; Pérez-Pulido, A.J.; Varela-López, A.; Quiles, J.L. NLRP3 inflammasome suppression improves longevity and prevents cardiac aging in male mice. Aging Cell 2020, 19, e13050. [Google Scholar] [CrossRef] [PubMed]
- Marneros, A.G. NLRP3 inflammasome blockade inhibits VEGF-A-induced age-related macular degeneration. Cell Rep. 2013, 4, 945–958. [Google Scholar] [CrossRef] [PubMed]
- Niu, L.; Zhang, S.; Wu, J.; Chen, L.; Wang, Y. Upregulation of NLRP3 Inflammasome in the Tears and Ocular Surface of Dry Eye Patients. PLoS ONE 2015, 10, e0126277. [Google Scholar] [CrossRef]
- Ozaki, E.; Campbell, M.; Doyle, S.L. Targeting the NLRP3 inflammasome in chronic inflammatory diseases: Current perspectives. J. Inflamm. Res. 2015, 8, 15. [Google Scholar]
- Park, B.; Jo, K.; Lee, T.G.; Hyun, S.W.; Kim, J.S.; Kim, C.S. Polydatin Inhibits NLRP3 Inflammasome in Dry Eye Disease by Attenuating Oxidative Stress and Inhibiting the NF-kappaB Pathway. Nutrients 2019, 11, 2792. [Google Scholar] [CrossRef]
- Pronin, A.; Pham, D.; An, W.; Dvoriantchikova, G.; Reshetnikova, G.; Qiao, J.; Kozhekbaeva, Z.; Reiser, A.E.; Slepak, V.Z.; Shestopalov, V.I. Inflammasome Activation Induces Pyroptosis in the Retina Exposed to Ocular Hypertension Injury. Front. Mol. Neurosci. 2019, 12, 36. [Google Scholar] [CrossRef]
- Saresella, M.; La Rosa, F.; Piancone, F.; Zoppis, M.; Marventano, I.; Calabrese, E.; Rainone, V.; Nemni, R.; Mancuso, R.; Clerici, M. The NLRP3 and NLRP1 inflammasomes are activated in Alzheimer’s disease. Mol. Neurodegener. 2016, 11, 23. [Google Scholar] [CrossRef]
- Seok, J.K.; Kang, H.C.; Cho, Y.Y.; Lee, H.S.; Lee, J.Y. Therapeutic regulation of the NLRP3 inflammasome in chronic inflammatory diseases. Arch. Pharm. Res. 2021, 44, 16–35. [Google Scholar] [CrossRef] [PubMed]
- Shao, B.Z.; Cao, Q.; Liu, C. Targeting NLRP3 Inflammasome in the Treatment of CNS Diseases. Front. Mol. Neurosci. 2018, 11, 320. [Google Scholar] [CrossRef] [PubMed]
- Shome, A.; Mugisho, O.O.; Niederer, R.L.; Rupenthal, I.D. Blocking the inflammasome: A novel approach to treat uveitis. Drug Discov. Today 2021, 26, 2839–2857. [Google Scholar] [CrossRef] [PubMed]
- Song, L.; Pei, L.; Yao, S.; Wu, Y.; Shang, Y. NLRP3 Inflammasome in Neurological Diseases, from Functions to Therapies. Front. Cell. Neurosci. 2017, 11, 63. [Google Scholar] [CrossRef] [PubMed]
- Tan, M.-S.; Yu, J.-T.; Jiang, T.; Zhu, X.-C.; Tan, L. The NLRP3 inflammasome in Alzheimer’s disease. Mol. Neurobiol. 2013, 48, 875–882. [Google Scholar] [CrossRef] [PubMed]
- Vakrakou, A.G.; Boiu, S.; Ziakas, P.D.; Xingi, E.; Boleti, H.; Manoussakis, M.N. Systemic activation of NLRP3 inflammasome in patients with severe primary Sjogren’s syndrome fueled by inflammagenic DNA accumulations. J. Autoimmun. 2018, 91, 23–33. [Google Scholar] [CrossRef]
- Wang, K.; Yao, Y.; Zhu, X.; Zhang, K.; Zhou, F.; Zhu, L. Amyloid beta induces NLRP3 inflammasome activation in retinal pigment epithelial cells via NADPH oxidase- and mitochondria-dependent ROS production. J. Biochem. Mol. Toxicol. 2017, 31, e21887. [Google Scholar] [CrossRef]
- Wang, W.; Wang, X.; Chun, J.; Vilaysane, A.; Clark, S.; French, G.; Bracey, N.A.; Trpkov, K.; Bonni, S.; Duff, H.J.; et al. Inflammasome-independent NLRP3 augments TGF-beta signaling in kidney epithelium. J. Immunol. 2013, 190, 1239–1249. [Google Scholar] [CrossRef]
- Wang, Y.; Liu, X.; Shi, H.; Yu, Y.; Yu, Y.; Li, M.; Chen, R. NLRP3 inflammasome, an immune-inflammatory target in pathogenesis and treatment of cardiovascular diseases. Clin. Transl. Med. 2020, 10, 91–106. [Google Scholar] [CrossRef]
- Yerramothu, P.; Vijay, A.K.; Willcox, M.D.P. Inflammasomes, the eye and anti-inflammasome therapy. Eye 2018, 32, 491–505. [Google Scholar] [CrossRef]
- Zhou, K.R.; Shi, L.G.; Wang, Y.; Chen, S.; Zhang, J.M. Recent Advances of the NLRP3 Inflammasome in Central Nervous System Disorders. J. Immunol. Res. 2016, 2016, 9238290. [Google Scholar] [CrossRef] [PubMed]
- Zmora, N.; Levy, M.; Pevsner-Fishcer, M.; Elinav, E. Inflammasomes and intestinal inflammation. Mucosal Immunol. 2017, 10, 865–883. [Google Scholar] [CrossRef] [PubMed]
- Vujosevic, S.; Bini, S.; Midena, G.; Berton, M.; Pilotto, E.; Midena, E. Hyperreflective intraretinal spots in diabetics without and with nonproliferative diabetic retinopathy: An in vivo study using spectral domain OCT. J. Diabetes Res. 2013, 2013, 1–5. [Google Scholar] [CrossRef] [PubMed]
- Vujosevic, S.; Bini, S.; Torresin, T.; Berton, M.; Midena, G.; Parrozzani, R.; Martini, F.; Pucci, P.; Daniele, A.R.; Cavarzeran, F. Hyperreflective retinal spots in normal and diabetic eyes: B-scan and en face spectral domain optical coherence tomography evaluation. Retina 2017, 37, 1092–1103. [Google Scholar] [CrossRef] [PubMed]
Pathology | Timepoint | Incidence per Treatment Group (Number of Animals and Percentage of Total Number) | |
---|---|---|---|
Vehicle (n = 8) | Tonabersat (n = 12) | ||
Vessel dilation | Day 2 | 8 (100%) | 2 (16.7%) **** |
Vessel tortuosity | Day 2 | 7 (87.5%) | 4 (33.3%) **** |
Vessel beading | Day 2 | 7 (87.5%) | 6 (50%) **** |
Hyperreflective foci | Day 2 | 3 (37.5%) | 0 (0%) **** |
Sub-retinal fluid accumulation | Day 7 | 2 (25%) | 0 (0%) **** |
Molecular Marker | Role | Antibody | Antibody Type | Working Dilution | Source |
---|---|---|---|---|---|
Connexin43 | Target protein | Rabbit polyclonal | Primary | 1:2000 | Sigma Aldrich, St Louis, MO, USA #C6219 |
GFAP-Cy3 | Müller cell and astrocyte marker | Mouse monoclonal | Primary | 1:1000 | Sigma Aldrich, #C9205 |
Iba1 | Microglia marker | Rabbit monoclonal | Primary | 1:2000 | Abcam plc, Cambridge, UK #ab178846 |
NLRP3 | Inflammasome marker | Goat polyclonal | Primary | 1:500 | Abcam plc, #ab4207 |
Cleaved caspase-1 | Inflammasome marker | Rabbit polyclonal | Primary | 1:50 | Invitrogen, Auckland, New Zealand #PA5-38099 |
Isolectin-B4-A594 | Blood vessel stain | Griffonia simplicifolia | Primary | 1:100 | Molecular Probes, Eugene, OR, USA #I-21413 |
PLVAP | Marker of leaky blood vessels | Mouse monoclonal | Primary | 1:1000 | Abcam plc, #ab27853 |
NF-κB | Inflammation marker | Rabbit polyclonal | Primary | 1:1000 | Abcam plc, #ab16502 |
Goat anti-rabbit Alexa Fluor 488 | - | Goat polyclonal | Secondary | 1:500 | Invitrogen, #A11034 |
Donkey anti-rabbit Alexa Fluor 488 | - | Donkey polyclonal | Secondary | 1:500 | Invitrogen, #A21206 |
Donkey anti-goat Cy3 | - | Donkey polyclonal | Secondary | 1:500 | Jackson Immuno Research Laboratories Inc., Westgrove, PA, USA #705-165-147 |
Disclaimer/Publisher’s Note: The statements, opinions and data contained in all publications are solely those of the individual author(s) and contributor(s) and not of MDPI and/or the editor(s). MDPI and/or the editor(s) disclaim responsibility for any injury to people or property resulting from any ideas, methods, instructions or products referred to in the content. |
© 2023 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
Share and Cite
Mugisho, O.O.; Aryal, J.; Shome, A.; Lyon, H.; Acosta, M.L.; Green, C.R.; Rupenthal, I.D. Orally Delivered Connexin43 Hemichannel Blocker, Tonabersat, Inhibits Vascular Breakdown and Inflammasome Activation in a Mouse Model of Diabetic Retinopathy. Int. J. Mol. Sci. 2023, 24, 3876. https://doi.org/10.3390/ijms24043876
Mugisho OO, Aryal J, Shome A, Lyon H, Acosta ML, Green CR, Rupenthal ID. Orally Delivered Connexin43 Hemichannel Blocker, Tonabersat, Inhibits Vascular Breakdown and Inflammasome Activation in a Mouse Model of Diabetic Retinopathy. International Journal of Molecular Sciences. 2023; 24(4):3876. https://doi.org/10.3390/ijms24043876
Chicago/Turabian StyleMugisho, Odunayo O., Jyoti Aryal, Avik Shome, Heather Lyon, Monica L. Acosta, Colin R. Green, and Ilva D. Rupenthal. 2023. "Orally Delivered Connexin43 Hemichannel Blocker, Tonabersat, Inhibits Vascular Breakdown and Inflammasome Activation in a Mouse Model of Diabetic Retinopathy" International Journal of Molecular Sciences 24, no. 4: 3876. https://doi.org/10.3390/ijms24043876