RNA Interference-Mediated Silencing ofAtp6iPrevents Both Periapical Bone Erosion and Inflammation in the Mouse Model of Endodontic Disease

ABSTRACTDental caries is one of the most prevalent infectious diseases in the United States, affecting approximately 80% of children and the majority of adults. Dental caries may lead to endodontic disease, where the bacterial infection progresses to the root canal system of the tooth, leading to periapical inflammation, bone erosion, severe pain, and tooth loss. Periapical inflammation may also exacerbate inflammation in other parts of the body. Although conventional clinical therapies for this disease are successful in approximately 80% of cases, there is still an urgent need for increased efficacy of treatment. In this study, we applied a novel gene-therapeutic approach using recombinant adeno-associated virus (AAV)-mediated Atp6i RNA interference (RNAi) knockdown ofAtp6i/TIRC7gene expression to simultaneously target periapical bone resorption and periapical inflammation. We found thatAtp6iinhibition impaired osteoclast functionin vitroandin vivoand decreased the number of T cells in the periapical lesion. Notably, AAV-mediatedAtp6i/TIRC7knockdown gene therapy reduced bacterial infection-stimulated bone resorption by 80% in the mouse model of endodontic disease. Importantly,Atp6i+/−mice with haploinsufficiency ofAtp6iexhibited protection similar to that in mice with bacterial infection-stimulated bone erosion and periapical inflammation, which confirms the potential therapeutic effect of AAV-small hairpin RNA (shRNA)-Atp6i/TIRC7. Our results demonstrate that AAV-mediatedAtp6i/TIRC7knockdown in periapical tissues can inhibit endodontic disease development, bone resorption, and inflammation, indicating for the first time that this potential gene therapy may significantly improve the health of those who suffer from endodontic disease.

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Inhibiting Periapical Lesions through AAV-RNAi Silencing of Cathepsin K

Journal of Dental Research ◽

10.1177/0022034512468757 ◽

2012 ◽

Vol 92 (2) ◽

pp. 180-186 ◽

Cited By ~ 32

Author(s):

B. Gao ◽

W. Chen ◽

L. Hao ◽

G. Zhu ◽

S. Feng ◽

...

Keyword(s):

Bone Resorption ◽

Dental Caries ◽

Bone Destruction ◽

Cathepsin K ◽

The Body ◽

Periapical Lesion ◽

Rnai Silencing ◽

Endodontic Disease ◽

Periapical Inflammation

Dental caries, one of the most prevalent infectious diseases worldwide, affects approximately 80% of children and the majority of adults. Dental caries may result in endodontic disease, leading to dental pulp necrosis, periapical inflammation and bone resorption, severe pain, and tooth loss. Periapical inflammation may also increase inflammation in other parts of the body. Although many studies have attempted to develop therapies for this disease, there is still an urgent need for effective treatments. In this study, we applied a novel gene therapeutic approach using recombinant adeno-associated virus (AAV)-mediated RNAi knockdown of Cathepsin K (Ctsk) gene expression, to target osteoclasts and periapical bone resorption in a mouse model. We found that AAV-sh-Cathepsin K (AAV-sh-Ctsk) impaired osteoclast function in vivo and furthermore reduced bacterial infection-stimulated bone resorption by 88%. Reduced periapical lesion size was accompanied by decreases in mononuclear leukocyte infiltration and inflammatory cytokine expression. Our study shows that AAV-RNAi silencing of Cathepsin K in periapical tissues can significantly reduce endodontic disease development, bone destruction, and inflammation in the periapical lesion. This is the first demonstration that AAV-mediated RNAi knockdown gene therapy may significantly reduce the severity of endodontic disease.

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SIGNIFICANCE AND NEED OF TREATMENT OF PERIAPICAL LESION

INDIAN DRUGS ◽

10.53879/id.51.07.p0005 ◽

2014 ◽

Vol 51 (07) ◽

pp. 5-13

Author(s):

S.R Pattan ◽

◽

P.S Bykod ◽

P.U. Shetkar ◽

S.V. Pattewar ◽

...

Keyword(s):

Dental Caries ◽

Infectious Diseases ◽

Dental Pulp ◽

Severe Pain ◽

Tooth Loss ◽

The Body ◽

Periapical Lesion ◽

Endodontic Disease ◽

Periapical Inflammation ◽

Effective Treatments

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Long-term RNA interference gene therapy in a dominant retinitis pigmentosa mouse model

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.1112758108 ◽

2011 ◽

Vol 108 (45) ◽

pp. 18476-18481 ◽

Cited By ~ 27

Author(s):

L. Jiang ◽

H. Zhang ◽

A. M. Dizhoor ◽

S. E. Boye ◽

W. W. Hauswirth ◽

...

Keyword(s):

Gene Therapy ◽

Rna Interference ◽

Mouse Model ◽

Retinitis Pigmentosa

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Silencing of Ac45 Simultaneously Inhibits Osteoclast-Mediated Bone Resorption and Attenuates Dendritic Cell-Mediated Inflammation through Impairing Acidification and Cathepsin K Secretion

Infection and Immunity ◽

10.1128/iai.00436-20 ◽

2020 ◽

Vol 89 (1) ◽

pp. e00436-20

Author(s):

Wenbin Yang ◽

Zheng Zhu ◽

Longjiang Li ◽

Abigail McVicar ◽

Ning Gao ◽

...

Keyword(s):

Dendritic Cell ◽

Bone Resorption ◽

Bone Erosion ◽

Bone Destruction ◽

Cathepsin K ◽

Osteoclast Formation ◽

Sequencing Analysis ◽

Necrosis Factor Alpha ◽

K Secretion ◽

Endodontic Disease

ABSTRACTEndodontic disease is characterized by inflammation and destruction of periapical tissues, leading to severe bone resorption and tooth loss. ATP6AP1 (Ac45) has been implicated in human immune diseases, yet the mechanism underlying how Ac45 regulates immune response and reaction in inflammatory diseases remains unknown. We generated endodontic disease mice through bacterial infection as an inflammatory disease model and used adeno-associated virus (AAV)-mediated Ac45 RNA interference knockdown to study the function of Ac45 in periapical inflammation and bone resorption. We demonstrated that the AAV small hairpin RNA targeting Ac45 (AAV-sh-Ac45) impaired cellular acidification, extracellular acidification, and bone resorption. Our results showed that local delivery of AAV-sh-Ac45 in periapical tissues in bacterium-induced inflammatory lesions largely reduced bone destruction, inhibited inflammation, and dramatically reduced mononuclear immune cells. T-cell, macrophage, and dendritic cell infiltration in the periapical lesion was dramatically reduced, and the periodontal ligament was protected from inflammation-induced destruction. Furthermore, AAV-sh-Ac45 significantly reduced osteoclast formation and the expression of proinflammatory cytokines, such as tumor necrosis factor alpha, interleukin-10 (IL-10), IL-12, IL-1α, IL-6, and IL-17. Interestingly, AAV-sh-Ac45 impaired mature cathepsin K secretion more significantly than that by AAV-sh-C1 and AAV-sh-CtsK. Unbiased genome-wide transcriptome sequencing analysis of Ctsk−/− dendritic cells stimulated with lipopolysaccharide demonstrated that the ablation of Ctsk dramatically reduced dendritic cell-mediated inflammatory signaling. Taken together, our results indicated that AAV-sh-Ac45 simultaneously inhibits osteoclast-mediated bone resorption and attenuates dendritic cell-mediated inflammation through impairing acidification and cathepsin K secretion. Thus, Ac45 may be a novel target for therapeutic approaches to attenuate inflammation and bone erosion in endodontic disease and other inflammation-related osteolytic diseases.

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