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Calcitriol prevents SARS-CoV spike-induced inflammation in human trophoblasts through downregulating ACE2 and TMPRSS2 expression

Vargas-Castro et al., The Journal of Steroid Biochemistry and Molecular Biology, doi:10.1016/j.jsbmb.2024.106625 (date from preprint)
Nov 2023  
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Vitamin D for COVID-19
8th treatment shown to reduce risk in October 2020, now with p < 0.00000000001 from 122 studies, recognized in 9 countries.
No treatment is 100% effective. Protocols combine treatments.
5,100+ studies for 111 treatments. c19early.org
In Vitro study showing that calcitriol prevents SARS-CoV-2 spike-induced inflammation in human trophoblasts through downregulation of ACE2 and TMPRSS2. Authors demonstrated that calcitriol significantly reduced expression of ACE2, TMPRSS2, and other renin-angiotensin system (RAS) components in both primary syncytiotrophoblasts and HTR-8/SVneo first-trimester trophoblast cells. Calcitriol treatment prevented spike protein-induced increases in inflammatory cytokines (TNFα, IL-1β, IL-6, IL-8, MCP-1) and reduced ACE1/ACE2 ratio. Effects were observed at concentrations as low as 0.1 nM calcitriol. The study suggests vitamin D may help prevent placental SARS-CoV-2 infection through multiple mechanisms: reducing viral entry receptor expression, decreasing inflammation, and modulating RAS components. Immunofluorescence confirmed reduced ACE2 protein levels and IL-6 expression with calcitriol treatment.
23 preclinical studies support the efficacy of vitamin D for COVID-19:
Vitamin D has been identified by the European Food Safety Authority (EFSA) as having sufficient evidence for a causal relationship between intake and optimal immune system function22-25. Vitamin D inhibits SARS-CoV-2 replication in vitro12,19, mitigates lung inflammation, damage, and lethality in mice with an MHV-3 model for β-CoV respiratory infections12,19, reduces SARS-CoV-2 replication in nasal epithelial cells via increased type I interferon expression15, downregulates proinflammatory cytokines IL-1β and TNF-α in SARS-CoV-2 spike protein-stimulated cells11, attenuates nucleocapsid protein-induced hyperinflammation by inactivating the NLRP3 inflammasome through the VDR-BRCC3 signaling pathway16, may be neuroprotective by protecting the blood-brain barrier, reducing neuroinflammation, and via immunomodulatory effects26, downregulates ACE2 and TMPRSS2 in human trophoblasts and minimizes spike protein-induced inflammation14, minimizes platelet aggregation mediated by SARS-CoV-2 spike protein via inhibiting integrin αIIbβ3 outside-in signaling10, and improves regulatory immune cell levels and control of proinflammatory cytokines in severe COVID-1927. Calcifediol inhibits SARS-CoV-2 papain-like protease (PLpro), a critical enzyme for viral replication9. Symptomatic COVID-19 is associated with a lower frequency of natural killer (NK) cells and vitamin D has been shown to improve NK cell activity28,29.
Vargas-Castro et al., 7 Nov 2023, peer-reviewed, 6 authors. Contact: lorenza.diazn@incmnsz.mx (corresponding author), lorenza.diazn@incmnsz.mx (corresponding author), rrvvcc14@gmail.com, janice.garciaq@incmnsz.mx, euclides.avilac@incmnsz.mx, fernando.larreag@incmnsz.mx, nut.aolmos@gmail.com.
In Vitro studies are an important part of preclinical research, however results may be very different in vivo.
This PaperVitamin DAll
Calcitriol Downregulates ACE1/ACE2, Renin and TMPRSS2 Gene Expression in the Human Placenta
Rafael Vargas-Castro, Andrea Olmos-Ortiz, Janice García-Quiroz, Euclides Avila, Fernando Larrea, Lorenza Díaz
doi:10.20944/preprints202311.0402.v1
Angiotensin-converting enzyme (ACE)1, ACE2, and renin are components of the renin-angiotensin system (RAS), which regulates blood pressure. ACE2 also serves as a receptor for SARS-CoV-2 and together with the transmembrane serine protease 2 (TMPRSS-2), mediates viral cell-endocytosis. As the placenta expresses all these factors, it acts as a target for SARS-CoV-2 and also as a source of blood pressure modulators. An ACE1/ACE2 ratio imbalance can lead to RAS dysregulation and a bad prognosis in COVID-19 patients. Calcitriol, the vitamin D active metabolite, negatively regulates RAS, reduces inflammation, and enhances antiviral immunity, thereby playing a protective role against COVID-19 severity. Placental calcitriol has been inversely correlated with maternal blood pressure; however, its regulatory role in RAS components and SARS-CoV-2 receptors within the fetomaternal unit has been barely explored. Therefore, we investigated the effects of calcitriol on placental RAS components. Calcitriol downregulated ACE1, ACE2, TMPRSS-2, and renin gene expression in cultured syncytiotrophoblasts and the extravillous trophoblast cell line HTR-8/SVneo. The ACE1/ACE2 ratio was also downregulated by calcitriol. Similar results were obtained in syncytiotrophoblasts treated with calcidiol, the precursor of calcitriol. Altogether, these results support that vitamin D is essential in restricting SARS-CoV-2 placental infection while helping to regulate maternal blood pressure during pregnancy.
Funding: This study was supported by the Annual Acquisition Program from the Departamento de Biología de la Reproducción Dr. Carlos Gual Castro, Instituto Nacional de Ciencias Médicas y Nutrición Salvador Zubirán, REF 2221 to L.D. The funders had no role in the study design, analysis and interpretation of the data, writing of the manuscript or the decision to submit the article for publication. Institutional Review Board Statement: Not applicable. Informed Consent Statement: Not applicable Conflicts of Interest: The authors declare no conflict of interest.
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