Periodontitis exacerbates pulmonary hypertension by promoting IFNγ⁺ T cell infiltration in mice

Abstract

Uncovering the risk factors of pulmonary hypertension and its mechanisms is crucial for the prevention and treatment of the disease. In the current study, we showed that experimental periodontitis, which was established by ligation of molars followed by orally smearing subgingival plaques from patients with periodontitis, exacerbated hypoxia-induced pulmonary hypertension in mice. Mechanistically, periodontitis dysregulated the pulmonary microbiota by promoting ectopic colonization and enrichment of oral bacteria in the lungs, contributing to pulmonary infiltration of interferon gamma positive (IFNγ⁺) T cells and aggravating the progression of pulmonary hypertension. In addition, we identified Prevotella zoogleoformans as the critical periodontitis-associated bacterium driving the exacerbation of pulmonary hypertension by periodontitis, and the exacerbation was potently ameliorated by both cervical lymph node excision and IFNγ neutralizing antibodies. Our study suggests a proof of concept that the combined prevention and treatment of periodontitis and pulmonary hypertension are necessary.

References

1. Kylhammar, D.et al.A comprehensive risk stratification at early follow-up determines prognosis in pulmonary arterial hypertension.Eur. Heart J. 39, 4175-4181 (2018).
2. Frost, A.et al.Diagnosis of pulmonary hypertension. Eur. Respir. J. 53, https://doi.org/10.1183/13993003.01904-2018 (2019).
3. Hassoun P. M.& Taichman, D. B. Pulmonary arterial hypertension.N. Engl. J. Med. 385, 2361-2376 (2021).
4. Humbert, M.et al.Sotatercept for the treatment of pulmonary arterial hypertension.N. Engl. J. Med. 384, 1204-1215 (2021).
5. Chen, J.et al.Microbiome and metabolome dysbiosis of the gut-lung axis in pulmonary hypertension.Microbiol. Res. 265, 127205(2022).
6. Chen Y. H., Yuan W., Meng L. K., Zhong, J. C. & Liu, X. Y. The role and mechanism of gut microbiota in pulmonary arterial hypertension. Nutrients 14, https://doi.org/10.3390/nu14204278 (2022).
7. Huang L., Zhang H., Liu Y.& Long, Y. The role of gut and airway microbiota in pulmonary arterial hypertension.Front Microbiol. 13, 929752(2022).
8. Oliveira S. D.Insights on the gut-mesentery-lung axis in pulmonary arterial hypertension: a poorly investigated crossroad.Arterioscler. Thromb. Vasc. Biol. 42, 516-526 (2022).
9. Wu P., Zhu T., Tan Z., Chen S.& Fang, Z. Role of gut microbiota in pulmonary arterial hypertension.Front Cell Infect. Microbiol. 12, 812303(2022).
10. Zhang, C.et al.Altered airway microbiota composition in patients with pulmonary hypertension.Hypertension 76, 1589-1599 (2020).
11. Kim, S.et al.Altered gut microbiome profile in patients with pulmonary arterial hypertension.Hypertension 75, 1063-1071 (2020).
12. Karoor, V.et al. The short-chain fatty acid butyrate attenuates pulmonary vascular remodeling and inflammation in hypoxia-induced pulmonary hypertension. Int. J. Mol. Sci. 22, https://doi.org/10.3390/ijms22189916 (2021).
13. Huang, Y.et al.Gut microbial metabolite trimethylamine N-oxide aggravates pulmonary hypertension.Am. J. Respir. Cell Mol. Biol. 66, 452-460 (2022).
14. Moutsoglou, D. M.et al.Pulmonary arterial hypertension patients have a proinflammatory gut microbiome and altered circulating microbial metabolites.Am. J. Respir. Crit. Care Med. 207, 740-756 (2023).
15. Sanada, T. J.et al.Gut microbiota modification suppresses the development of pulmonary arterial hypertension in an SU5416/hypoxia rat model.Pulmonary Circ. 10, 2045894020929147-2045894020929147 (2020).
16. Gaeckle N. T., Pragman A. A., Pendleton K. M., Baldomero A. K.& Criner, G. J. The oral-lung axis: the impact of oral health on lung health.Respir Care 65, 1211-1220 (2020).
17. Dong, J.et al.Relationships between oral microecosystem and respiratory diseases.Front Mol. Biosci. 8, 718222(2021).
18. Segal, L. N.et al.Enrichment of the lung microbiome with oral taxa is associated with lung inflammation of a Th17 phenotype.Nat. Microbiol. 1, 16031(2016).
19. Mammen, M. J., Scannapieco, F. A.& Sethi, S. Oral-lung microbiome interactions in lung diseases.Periodontol 2000 83, 234-241 (2020).
20. Pu C. Y., Seshadri M., Manuballa S.& Yendamuri, S. The Oral Microbiome and Lung Diseases.Current Oral Health Reports 7, 79-86 (2020).
21. Hajishengallis G.Interconnection of periodontal disease and comorbidities: Evidence, mechanisms, and implications.Periodontol 2000 89, 9-18 (2022).
22. Hajishengallis, G. & Chavakis, T. Local and systemic mechanisms linking periodontal disease and inflammatory comorbidities. Nat Rev Immunol, 1-15, (2021).
23. Dorfmüller P., Perros F., Balabanian K.& Humbert, M. Inflammation in pulmonary arterial hypertension.European Respiratory Journal 22, 358(2003).
24. Schermuly R. T., Ghofrani H. A., Wilkins M. R.& Grimminger, F. Mechanisms of disease: pulmonary arterial hypertension.Nat Rev Cardiol 8, 443-455 (2011).
25. Wang, R.-R.et al.Immunity and inflammation in pulmonary arterial hypertension: From pathophysiology mechanisms to treatment perspective.Pharmacological Research 180, 106238(2022).
26. Rabinovitch M., Guignabert C., Humbert M.& Nicolls, M. R. Inflammation and immunity in the pathogenesis of pulmonary arterial hypertension.Circ Res. 115, 165-175 (2014).
27. Ryan J. J.& Archer, S. L. The right ventricle in pulmonary arterial hypertension: disorders of metabolism, angiogenesis and adrenergic signaling in right ventricular failure.Circ Res. 115, 176-188 (2014).
28. Tuder R. M., Groves B., Badesch D. B.& Voelkel, N. F. Exuberant endothelial cell growth and elements of inflammation are present in plexiform lesions of pulmonary hypertension.Am. J. Pathol. 144, 275-285 (1994).
29. Cool C. D., Kennedy D., Voelkel N. F.& Tuder, R. M. Pathogenesis and evolution of plexiform lesions in pulmonary hypertension associated with scleroderma and human immunodeficiency virus infection.Hum. Pathol. 28, 434-442 (1997).
30. Zhang, J. et al. Differential oral microbial input determines two microbiota pneumo-types associated with health status. Adv. Sci. (Weinh) 9, e2203115 (2022).
31. Sanz, M.et al.Periodontitis and cardiovascular diseases: consensus report.J. Clin. Periodontol. 47, 268-288 (2020).
32. Herrera, D.et al.Association between periodontal diseases and cardiovascular diseases, diabetes and respiratory diseases: Consensus report of the Joint Workshop by the European Federation of Periodontology (EFP) and the European arm of the World Organization of Family Doctors (WONCA Europe).J. Clin. Periodontol. 50, 819-841 (2023).
33. Czesnikiewicz-Guzik, M. et al. Causal association between periodontitis and hypertension: evidence from Mendelian randomization and a randomized controlled trial of non-surgical periodontal therapy.Eur. Heart J. 40, 3459-3470 (2019).
34. Chen, B.-Y.et al.Roles of oral microbiota and oral-gut microbial transmission in hypertension.J. Adv. Res. 43, 147-161 (2023).
35. Hajishengallis G.Periodontitis: from microbial immune subversion to systemic inflammation.Nat. Rev. Immunol. 15, 30-44 (2015).
36. Lourenςo T. G.B., Spencer, S. J., Alm, E. J. & Colombo, A. P. V. Defining the gut microbiota in individuals with periodontal diseases: an exploratory study.J. Oral Microbiol. 10, 1487741(2018).
37. Kitamoto, S.et al.The Intermucosal Connection between the Mouth and Gut in Commensal Pathobiont-Driven Colitis. Cell, https://doi.org/10.1016/j.cell.2020.05.048(2020).
38. Bui, F. Q.et al.Association between periodontal pathogens and systemic disease.Biomed. J. 42, 27-35 (2019).
39. Shen, X.et al.Ectopic colonization and immune landscapes of periodontitis microbiota in germ-free mice with streptozotocin-induced type 1 diabetes mellitus. Front. Microbiol. 13, https://doi.org/10.3389/fmicb.2022.889415(2022).
40. Atarashi, K.et al.Ectopic colonization of oral bacteria in the intestine drives T(H)1 cell induction and inflammation.Science 358, 359-365 (2017).
41. Blasco-Baque, V. et al. Periodontitis induced by Porphyromonas gingivalis drives periodontal microbiota dysbiosis and insulin resistance via an impaired adaptive immune response.Gut. 66, 872-885 (2017).
42. Zhang, C.et al.Airway delivery of Streptococcus salivarius is sufficient to induce experimental pulmonary hypertension in rats.Br. J. Pharmacol. 180, 2102-2119 (2023).
43. Swain S. D., Siemsen D. W., Pullen R. R.& Han, S. CD4+ T cells and IFN-γ are required for the development of pneumocystis-associated pulmonary hypertension.Am. J. Pathol. 184, 483-493 (2014).
44. Tomaszewski, M.et al. Overexpression of PD-1 on peripheral blood lymphocytes in patients with idiopathic pulmonary arterial hypertension and its association with high viral loads of epstein-barr virus and poor clinical parameters. J. Clin. Med. 9, https://doi.org/10.3390/jcm9061966 (2020).
45. Li S., Ma X., Xie J., Yan X.& Sun, W. MicroRNA-206, IL-4, IL-13, and INF-γ levels in lung tissue and plasma are increased by the stimulation of particulate matter with a diameter of ≤2.5?μm, and are associated with the poor prognosis of asthma induced pulmonary arterial hypertension patients.Clin. Experimental Hypertension 43, 181-188 (2021).
46. Liu, S.-F.et al.Pulmonary hypertension: Linking inflammation and pulmonary arterial stiffening. Front. Immunol. 13, https://doi.org/10.3389/fimmu.2022.959209(2022).
47. George, P. M.et al.Evidence for the involvement of type I interferon in pulmonary arterial hypertension.Circ. Res. 114, 677-688 (2014).
48. Papani R., Duarte A. G., Lin Y. L., Kuo Y. F.& Sharma, G. Pulmonary arterial hypertension associated with interferon therapy: a population-based study.Multidiscip. Respir. Med. 12, 1(2017).
49. Glasner, A.et al. NKp46 receptor-mediated interferon-γ production by natural killer cells increases fibronectin 1 to alter tumor architecture and control metastasis. Immunity 48, 107-119.e104 (2018).
50. Ma, X. X.et al.Ncor1 deficiency promotes osteoclastogenesis and exacerbates periodontitis.J. Dent. Res. 102, 72-81 (2023).
51. Abe T.& Hajishengallis, G. Optimization of the ligature-induced periodontitis model in mice.J. Immunol. Methods 394, 49-54 (2013).
52. Bai L., Chen B.-Y., Liu Y., Zhang W.-C.& Duan, S.-Z. A mouse periodontitis model with humanized oral bacterial community. Front. Cell. Infect. Microbiol. 12, https://doi.org/10.3389/fcimb.2022.842845(2022).
53. Zhou, L. J.et al.Oral pathobionts promote MS-like symptoms in mice.J. Dental Res. 102, 217-226 (2022).
54. van Zwam, M.et al. Surgical excision of CNS‐draining lymph nodes reduces relapse severity in chronic‐relapsing experimental autoimmune encephalomyelitis.J. Pathol. 217, 543-551 (2008).
55. Zhu, Z.et al.Echocardiographic assessment of right ventricular function in experimental pulmonary hypertension.Pulm Circ. 9, 2045894019841987(2019).
56. Thibault, H. B.et al.Noninvasive assessment of murine pulmonary arterial pressure.Circ.: Cardiovas. Imaging 3, 157-163 (2010).
57. Du, L.-J.et al.NCOR1 maintains the homeostasis of vascular smooth muscle cells and protects against aortic aneurysm.Cell Death Differ. 30, 618-631 (2022).
58. Handoko, M. L.et al.A refined radio-telemetry technique to monitor right ventricle or pulmonary artery pressures in rats: a useful tool in pulmonary hypertension research.Pflugers Arch. 455, 951-959 (2008).
59. Shu, T.et al.Inhibition of immunoglobulin E attenuates pulmonary hypertension.Nat. Cardiovas. Res. 1, 665-678 (2022).
60. Dong, T.et al.Prevotella intermedia aggravates subclinical hypothyroidism.J. Dental Res. 102, 814-824 (2023).
61. Nejman, D.et al.The human tumor microbiome is composed of tumor type-specific intracellular bacteria.Science 368, 973-980 (2020).
62. Jungblut M., Oeltze K., Zehnter I., Hasselmann, D. & Bosio, A. Standardized preparation of single-cell suspensions from mouse lung tissue using the gentleMACS Dissociator. J. Vis. Exp. https://doi.org/10.3791/1266 (2009).