Calcium channel blockers reduce severe fever with thrombocytopenia syndrome virus (SFTSV) related fatality

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Abstract

Severe fever with thrombocytopenia syndrome (SFTS), an emerging tick-borne infectious disease caused by a novel phlebovirus (SFTS virus, SFTSV), was listed among the top 10 priority infectious diseases by the World Health Organization due to its high fatality of 12%–50% and possibility of pandemic transmission. Currently, effective anti-SFTSV intervention remains unavailable. Here, by screening a library of FDA-approved drugs, we found that benidipine hydrochloride, a calcium channel blocker (CCB), inhibited SFTSV replication in vitro. Benidipine hydrochloride was revealed to inhibit virus infection through impairing virus internalization and genome replication. Further experiments showed that a broad panel of CCBs, including nifedipine, inhibited SFTSV infection. The anti-SFTSV effect of these two CCBs was further analyzed in a humanized mouse model in which CCB treatment resulted in reduced viral load and decreased fatality rate. Importantly, by performing a retrospective clinical investigation on a large cohort of 2087 SFTS patients, we revealed that nifedipine administration enhanced virus clearance, improved clinical recovery, and remarkably reduced the case fatality rate by >5-fold. These findings are highly valuable for developing potential host-oriented therapeutics for SFTS and other lethal acute viral infections known to be inhibited by CCBs in vitro.

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Data availability

The data that support the findings of this study are available from the corresponding authors upon request.

References

  1. 1.

    Paules, C. I., Marston, H. D., Bloom, M. E. & Fauci, A. S. Tickborne diseases — confronting a growing threat. N. Engl. J. Med. 379, 701–703 (2018).

  2. 2.

    Yu, X. J. et al. Fever with thrombocytopenia associated with a novel bunyavirus in China. N. Engl. J. Med. 364, 1523–1532 (2011).

  3. 3.

    Liu, Q., He, B., Huang, S. Y., Wei, F. & Zhu, X. Q. Severe fever with thrombocytopenia syndrome, an emerging tick-borne zoonosis. Lancet Infect. Dis. 14, 763–772 (2014).

  4. 4.

    Kim, K. H. et al. Severe fever with thrombocytopenia syndrome, South Korea, 2012. Emerg. Infect. Dis. 19, 1892–1894 (2013).

  5. 5.

    Takahashi, T. et al. The first identification and retrospective study of severe fever with thrombocytopenia syndrome in Japan. J. Infect. Dis. 209, 816–827 (2014).

  6. 6.

    Li, H. et al. Epidemiological and clinical features of laboratory-diagnosed severe fever with thrombocytopenia syndrome in China, 2011-17: a prospective observational study. Lancet Infect. Dis. 18, 1127–1137 (2018).

  7. 7.

    2017 Annual review of diseases prioritized under the Research and Development Blueprint (WHO Meeting report, World Health Organization, 2017).

  8. 8.

    Luo, L. M. et al. Haemaphysalis longicornis ticks as reservoir and vector of severe fever with thrombocytopenia syndrome virus in China. Emerg. Infect. Dis. 21, 1770–1776 (2015).

  9. 9.

    Zhuang, L. et al. Transmission of severe fever with thrombocytopenia syndrome virus by Haemaphysalis longicornis ticks, China. Emerg. Infect. Dis. 24, https://doi.org/10.3201/eid2405.151435 (2018).

  10. 10.

    Park, S. W. et al. Prevalence of severe fever with thrombocytopenia syndrome virus in Haemaphysalis longicornis ticks in South Korea. Ticks Tick. Borne Dis. 5, 975–977 (2014).

  11. 11.

    Tateno, M. et al. Molecular survey of arthropod-borne pathogens in ticks obtained from Japanese wildcats. Ticks Tick. Borne Dis. 6, 281–289 (2015).

  12. 12.

    Hammer, J. F., Emery, D., Bogema, D. R. & Jenkins, C. Detection of Theileria orientalis genotypes in Haemaphysalis longicornis ticks from southern Australia. Parasit. Vectors 8, 229 (2015).

  13. 13.

    Heath, A. Biology, ecology and distribution of the tick, Haemaphysalis longicornis Neumann (Acari: Ixodidae) in New Zealand. N. Z. Vet. J. 64, 10–20 (2016).

  14. 14.

    Yun, Y. et al. Phylogenetic analysis of severe fever with thrombocytopenia syndrome virus in South Korea and migratory bird routes between China, South Korea, and Japan. Am. J. Trop. Med. Hyg. 93, 468–474 (2015).

  15. 15.

    Li, Z. et al. Ecology of the tick-borne phlebovirus causing severe fever with thrombocytopenia syndrome in an endemic area of China. PLoS Negl. Trop. Dis. 10, e0004574 (2016).

  16. 16.

    Rainey, T., Occi, J. L., Robbins, R. G. & Egizi, A. Discovery of Haemaphysalis longicornis (Ixodida: Ixodidae) parasitizing a sheep in New Jersey, United States. J. Med. Entomol. 55, 757–759 (2018).

  17. 17.

    National Haemaphysalis longicornis (longhorned tick) Situation Report – August 29, 2018 (United States Department of Agriculture, 2018).

  18. 18.

    Liu, W. et al. Case-fatality ratio and effectiveness of ribavirin therapy among hospitalized patients in china who had severe fever with thrombocytopenia syndrome. Clin. Infect. Dis. 57, 1292–1299 (2013).

  19. 19.

    Tani, H. et al. Efficacy of T-705 (Favipiravir) in the treatment of infections with lethal severe fever with thrombocytopenia syndrome virus. mSphere 1, e00061–15 (2016).

  20. 20.

    Smee, D. F., Jung, K. H., Westover, J. & Gowen, B. B. 2’-Fluoro-2’-deoxycytidine is a broad-spectrum inhibitor of bunyaviruses in vitro and in phleboviral disease mouse models. Antivir. Res. 160, 48–54 (2018).

  21. 21.

    Richardson, F. C. et al. Quantification of 2’-fluoro-2’-deoxyuridine and 2’-fluoro-2’-deoxycytidine in DNA and RNA isolated from rats and woodchucks using LC/MS/MS. Chem. Res. Toxicol. 15, 922–926 (2002).

  22. 22.

    Baba, M. et al. Establishment of an antiviral assay system and identification of severe fever with thrombocytopenia syndrome virus inhibitors. Antivir. Chem. Chemother. 25, 83–89 (2017).

  23. 23.

    Yuan, S. et al. Screening of an FDA-approved drug library with a two-tier system identifies an entry inhibitor of severe fever with thrombocytopenia syndrome virus. Viruses 11, E385 (2019).

  24. 24.

    Barrows, N. J. et al. A Screen of FDA-approved drugs for inhibitors of Zika virus infection. Cell Host Microbe 20, 259–270 (2016).

  25. 25.

    Johansen, L. M. et al. A screen of approved drugs and molecular probes identifies therapeutics with anti-Ebola virus activity. Sci. Transl. Med 7, 290ra89 (2015).

  26. 26.

    Zhang, J. H., Chung, T. D. & Oldenburg, K. R. A simple statistical parameter for use in evaluation and validation of high throughput screening assays. J. Biomol. Screen. 4, 67–73 (1999).

  27. 27.

    Yuan, F. & Zheng, A. Entry of severe fever with thrombocytopenia syndrome virus. Virol. Sin. 32, 44–50 (2017).

  28. 28.

    Tani, H. et al. Characterization of glycoprotein-mediated entry of severe fever with thrombocytopenia syndrome virus. J. Virol. 90, 5292–5301 (2016).

  29. 29.

    Liu, J. et al. Single-particle tracking reveals the sequential entry process of the Bunyavirus severe fever with thrombocytopenia syndrome virus. Small 15, e1803788 (2019).

  30. 30.

    Hermann, L. L. & Coombs, K. M. Inhibition of reovirus by mycophenolic acid is associated with the M1 genome segment. J. Virol. 78, 6171–6179 (2004).

  31. 31.

    Yao, K., Nagashima, K. & Miki, H. Pharmacological, pharmacokinetic, and clinical properties of benidipine hydrochloride, a novel, long-acting calcium channel blocker. J. Pharm. Sci. 100, 243–261 (2006).

  32. 32.

    Zhao, W. & Zhen, J.-C. Analysis of the utilization of oral antihypertensive drugs in more than 40 hospitals from Beijing area during 2008-2011. China Pharm. 24, 506–509 (2013).

  33. 33.

    Li, X.-P., Yu, Z.-Q. & Liu, D. Analysis of the application of antihypertensive drugs in 34 hospitals of Wuhan city during 2011 to 2013. China Pharm. 20, 2748–2751 (2015).

  34. 34.

    Seino, H. et al. Effect of benidipine hydrochloride, a long-acting T-type calcium channel blocker, on blood pressure and renal function in hypertensive patients with diabetes mellitus. Anal. Switch. cilnidipine benidipine. Arzneim. 57, 526–531 (2007).

  35. 35.

    Hayashi, K. et al. Ca2+ channel subtypes and pharmacology in the kidney. Circ. Res. 100, 342–353 (2007).

  36. 36.

    Helton, T. D., Xu, W. & Lipscombe, D. Neuronal L-type calcium channels open quickly and are inhibited slowly. J. Neurosci. 25, 10247–10251 (2005).

  37. 37.

    Jin, C. et al. Pathogenesis of emerging severe fever with thrombocytopenia syndrome virus in C57/BL6 mouse model. Proc. Natl Acad. Sci. USA 109, 10053–10058 (2012).

  38. 38.

    Zhang, Y. Z. et al. Hemorrhagic fever caused by a novel Bunyavirus in China: pathogenesis and correlates of fatal outcome. Clin. Infect. Dis. 54, 527–533 (2012).

  39. 39.

    Gai, Z. T. et al. Clinical progress and risk factors for death in severe fever with thrombocytopenia syndrome patients. J. Infect. Dis. 206, 1095–1102 (2012).

  40. 40.

    McMullan, L. K. et al. A new phlebovirus associated with severe febrile illness in Missouri. N. Engl. J. Med. 367, 834–841 (2012).

  41. 41.

    Muehlenbachs, A. et al. Heartland virus-associated death in tennessee. Clin. Infect. Dis. 59, 845–850 (2014).

  42. 42.

    Fill, M. A. et al. Novel clinical and pathologic findings in a Heartland virus-associated death. Clin. Infect. Dis. 64, 510–512 (2017).

  43. 43.

    Shen, S. et al. A novel tick-borne phlebovirus, closely related to severe fever with thrombocytopenia syndrome virus and Heartland virus, is a potential pathogen. Emerg. Microbes Infect. 7, 95 (2018).

  44. 44.

    Scherbik, S. V. & Brinton, M. A. Virus-induced Ca2+ influx extends survival of west nile virus-infected cells. J. Virol. 84, 8721–8731 (2010).

  45. 45.

    Zamponi, G. W. Targeting voltage-gated calcium channels in neurological and psychiatric diseases. Nat. Rev. Drug Discov. 15, 19–34 (2016).

  46. 46.

    Elliott, W. J. & Ram, C. V. Calcium channel blockers. J. Clin. Hypertens. 13, 687–689 (2011).

  47. 47.

    Bagur, R. & Hajnoczky, G. Intracellular Ca2+ sensing: Its role in calcium homeostasis and signaling. Mol. Cell 66, 780–788 (2017).

  48. 48.

    Clapham, D. E. Calcium signaling. Cell 131, 1047–1058 (2007).

  49. 49.

    Zhou, Y., Frey, T. K. & Yang, J. J. Viral calciomics: interplays between Ca2+ and virus. Cell Calcium 46, 1–17 (2009).

  50. 50.

    Ma, G. et al. phage HIV-1 infection. J. Exp. Med. 200, 1337–1346 (2004).

  51. 51.

    Choi, J. et al. Association of hepatitis B virus polymerase with promyelocytic leukemia nuclear bodies mediated by the S100 family protein p11. Biochem Biophys. Res. Commun. 305, 1049–1056 (2003).

  52. 52.

    Perez, M., Craven, R. C. & de la Torre, J. C. The small RING finger protein Z drives arenavirus budding: implications for antiviral strategies. Proc. Natl Acad. Sci. USA 100, 12978–12983 (2003).

  53. 53.

    Lingappa, U. F. et al. Host-rabies virus protein-protein interactions as druggable antiviral targets. Proc. Natl Acad. Sci. USA 110, E861–E868 (2013).

  54. 54.

    Andrei, G. & De Clercq, E. Molecular approaches for the treatment of hemorrhagic fever virus infections. Antivir. Res. 22, 45–75 (1993).

  55. 55.

    Garcia, M. et al. Productive replication of Ebola virus is regulated by the c-Abl1 tyrosine kinase. Sci. Transl. Med. 4, 123ra24 (2012).

  56. 56.

    Sakurai, Y. et al. Ebola virus. Two-pore channels control Ebola virus host cell entry and are drug targets for disease treatment. Science 347, 995–998 (2015).

  57. 57.

    DeWald, L. E. et al. The calcium channel blocker Bepridil demonstrates efficacy in the murine model of Marburg virus disease. J. Infect. Dis. 218, S588–S591 (2018).

  58. 58.

    Lavanya, M., Cuevas, C. D., Thomas, M., Cherry, S. & Ross, S. R. siRNA screen for genes that affect Junin virus entry uncovers voltage-gated calcium channels as a therapeutic target. Sci. Transl. Med. 5, 204ra131 (2013).

  59. 59.

    Wang, S. et al. Screening of FDA-approved drugs for inhibitors of Japanese encephalitis virus infection. J. Virol. 91, e01055-17 (2017).

  60. 60.

    Park, S. Y. et al. Use of plasma therapy for severe fever with thrombocytopenia syndrome encephalopathy. Emerg. Infect. Dis. 22, 1306–1308 (2016).

  61. 61.

    Oh, W. S. et al. Effect of early plasma exchange on survival in patients with severe fever with thrombocytopenia syndrome: a multicenter study. Yonsei Med. J. 58, 867–871 (2017).

  62. 62.

    Nakamura, S. et al. Steroid pulse therapy in patients with encephalopathy associated with severe fever with thrombocytopenia syndrome. J. Infect. Chemother. 24, 389–392 (2018).

  63. 63.

    Zhang, Y. et al. Isolation, characterization, and phylogenic analysis of three new severe fever with thrombocytopenia syndrome bunyavirus strains derived from Hubei Province, China. Virol. Sin. 32, 89–96 (2017).

  64. 64.

    Livonesi, M. C., Moro de Sousa, R. L. & Moraes Figueiredo, L. T. In vitro study of antiviral activity of mycophenolic acid on Brazilian orthobunyaviruses. Intervirology 50, 204–208 (2007).

  65. 65.

    Sepulveda, C. S. et al. Inhibition of Junin virus RNA synthesis by an antiviral acridone derivative. Antivir. Res. 93, 16–22 (2012).

  66. 66.

    Takhampunya, R., Ubol, S., Houng, H. S., Cameron, C. E. & Padmanabhan, R. Inhibition of dengue virus replication by mycophenolic acid and ribavirin. J. Gen. Virol. 87, 1947–1952 (2006).

  67. 67.

    Guo, S. et al. Oncological and genetic factors impacting PDX model construction with NSG mice in pancreatic cancer. FASEB J. 33, 873–884 (2019).

  68. 68.

    Ministry of Health PRC. Guideline for prevention and treatment of severe fever with thrombocytopenia syndrome (2010 vesion). Chin. J. Clin. Infect. Dis. 4, 193–194 (2011).

  69. 69.

    Cui, N. et al. Severe fever with thrombocytopenia syndrome bunyavirus-related human encephalitis. J. Infect. 70, 52–59 (2015).

  70. 70.

    Ding, S. et al. Age is a critical risk factor for severe fever with thrombocytopenia syndrome. PLoS ONE 9, e111736 (2014).

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Acknowledgements

This work was supported by the Strategic Priority Research Program of the Chinese Academy of Sciences (XDB29010204), the National Natural Science Foundation of China (81825019, 81722041, 31770188, 81472005, 81473023 and 31500144), the National Science and Technology Major Project (2018ZX10101004001005), the National Key R&D Program of China (2016YFC1200400, 2018YFA0507201 and 2016YFC1201905), the Hundred Talents Program of the Chinese Academy of Sciences (K.P.), the Youth Innovation Promotion Association of the Chinese Academy of Sciences (2018367), the Special Major Program of Wuhan Institute of Virology (WIV-135-TP1), the State Key Laboratory of Virology Open Projects (2017IOV003), the Foundation for Innovative Research Groups of the National Natural Science Foundation of China (81621005), and the New Star Plan of Science and Technology of Beijing (Z171100001117089).

Author information

K.P. and W.L. conceived and supervised the study. H.L., L.-K.Z., K.P. and W.L. designed the experiments, analyzed the results, and wrote the manuscript. H.L., L.-K.Z., S.-F.L., W.-W.W., Y.-L.Z., Q.-L.X., K.D., Y.-Y.H., X.-T.Z., Y.-J.F., P.-H.Z., J.-Y.B. and K.P. performed the experiments. H.L., S.-F.Z., Z.-B.W., N.C., C.Y. and W.L. recruited the patients. H.L., L.-K.Z., S.-F.Z. and Q.-B.L. performed statistical analysis. G.-F.X. and F.D. contributed to the design of the study and data analysis. All authors had access to the study data, and reviewed and approved the final manuscript.

Correspondence to Wei Liu or Ke Peng.

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