key: cord-0275674-p43v3ihz authors: Jalkanen, J.; Pettila, V.; Karvonen, M.; Huttunen, T.; Mandelin, J.; Jalkanen, M.; Malmberg, M.; Elima, K.; Bellingan, G.; Hollmen, M.; Ranieri, V. M.; Jalkanen, S. title: Glucocorticoids inhibit type I IFN beta signaling and the upregulation of CD73 in human lung date: 2020-04-06 journal: nan DOI: 10.1101/2020.04.01.20049700 sha: b73d822cbac4f2102af0d987378355e111a40cfd doc_id: 275674 cord_uid: p43v3ihz Abstract Purpose: Glucocorticoids are widely used to treat acute respiratory distress syndrome (ARDS) despite its use is highly controversial based on randomized controlled trials and meta-analyses. As type I interferons (IFNs) are our first line of defense against severe viral respiratory infections, we explored whether glucocorticoids interfere with IFN signaling and whether their use associates to outcome of IFN beta treatment of ARDS. Methods: We performed a propensity-matched post-hoc-analysis using data from the recent randomized INTEREST trial comparing IFN beta 1a to placebo in ARDS patients. Based on the results of these analyses we utilized human lung tissue and human pulmonary endothelial cell cultures to investigate the effect of hydrocortisone on IFN nuclear signaling and the protein transcription of CD73, a molecule responsible for vascular integrity. Results: We found that hydrocortisone reduces the production, and prevents the nuclear translocation of IRF9, that is required for IFN beta-dependent signaling of multiple IFN-induced genes. In addition, hydrocortisone inhibits IFN beta dependent upregulation of CD73 in human lung tissue. Additionally, we found that use of glucocorticoids with IFN beta 1a was independently associated with increased mortality (OR 5.4, 95% CI 2.1-13.9, P< 0.001) in the INTEREST-trial. Conclusions: Glucocorticoids inhibit type I IFN beta signaling and the upregulation of CD73 in human lung. This provides the mechanistic basis for the harmful association of glucocorticoids in IFN beta treated patients in the INTEREST-trial. Most importantly, it strongly speaks against the use of glucocorticoids in viral-induced ARDS such as in the currently expanding corona virus outbreak. i n h u m a n l u n g . I F N b e t a -d e p e n d e n t u p r e g u l a t i o n o f C D 7 3 i n h u m a n l u n g t i s s u e . A d d i t i o n a l l y , w e f o u n d t h a t u s e o f g l u c o c o r t i c o i d s w i t h I F N b e t a -1 a w a s i n d e p e n d e n t l y a s s o c i a t e d w i t h i n c r e a s e d m o r t a l i t y ( O R 5 . 4 , 9 5 % C I 2 . 1 -1 3 . 9 , P < 0 . 0 0 1 ) i n t h e I N T E R E S T -t r i a l . C o n c l u s i o n s : G l u c o c o r t i c o i d s i n h i b i t t y p e I I F N b e t a s i g n a l i n g a n d t h e u p r e g u l a t i o n o f C D 7 3 i n h u m a n l u n g . T h i s p r o v i d e s t h e m e c h a n i s t i c b a s i s f o r t h e h a r m f u l a s s o c i a t i o n o f g l u c o c o r t i c o i d s i n I F N b e t a t r e a t e d p a t i e n t s i n t h e I N T E R E S T -t r i a l . M o s t i m p o r t a n t l y , i t s t r o n g l y s p e a k s a g a i n s t t h e u s e o f g l u c o c o r t i c o i d s i n v i r a l -i n d u c e d A R D S s u c h a s i n t h e c u r r e n t l y e x p a n d i n g c o r o n a v i r u s o u t b r e a k . ( 2 4 6 w o r d s ) K e y w o r d s : C D 7 3 , e n d o t h e l i u m , A R D S , i n t e r f e r o n , g l u c o c o r t i c o i d s All rights reserved. No reuse allowed without permission. author/funder, who has granted medRxiv a license to display the preprint in perpetuity. I n t r o d u c t i o n A c u t e r e s p i r a t o r y d i s t r e s s s y n d r o m e ( A R D S ) i s a c o n d i t i o n c h a r a c t e r i z e d b y p u l m o n a r y i n f l a m m a t i o n , d i f f u s e p u l m o n a r y e d e m a a n d r e f r a c t o r y h y p o x e m i a t h a t m a y c o m p l i c a t e e n d o t h e l i a l -a l v e o l a r b a r r i e r w i t h i n c r e a s e d p u l m o n a r y v a s c u l a r l e a k a g e [ 1 , 3 ] . E n d o g e n o u s p r o d u c t i o n o f t y p e I I F N s ( I F N b e t a a n d a l p h a ) i s o f o u t m o s t i m p o r t a n c e t o f i g h t a g a i n s t v i r a l a n d b a c t e r i a l i n f e c t i o n s [ 4 ] , s i n c e i t u p r e g u l a t e s c l u s t e r o f d i f f e r e n t i a t i o n 7 3 ( C D 7 3 ) e x p r e s s e d o n e n d o t h e l i u m c e l l s . C D 7 3 i s a n e n z y m e c o n t r o l l i n g e n d o t h e l i a l b a r r i e r f u n c t i o n a n d l e u c o c y t e r e c r u i t m e n t t o s i t e s o f i n f l a m m a t i o n v i a t h e p r o d u c t i o n o f a d e n o s i n e [ 5 -7 ] , a h i g h l y a n t i -i n f l a m m a t o r y s u b s t a n c e h a v i n g c a r d i o p r o t e c t i v e , n e u r o p r o t e c t i v e , v a s o d i l a t o r y a n d a n g i o g e n i c p r o p e r t i e s [ 8 ] . A p h a s e I I t r i a l s h o w e d t h a t u p r e g u l a t i o n o f C D 7 3 v i a i n t r a v e n o u s a d m i n i s t r a t i o n o f r e c o m b i n a n t h u m a n i n t e r f e r o n i s a s s o c i a t e d w i t h a r e d u c t i o n i n 2 c o n t r o l l e d p h a s e I I I t r i a l ( I N T E R E S T ) s h o w e d t h a t i n t r a v e n o u s r e c o m b i n a n t h u m a n i n t e r f e r o n b e t a -1 a d i d n o t i m p r o v e o u t c o m e i n p a t i e n t s w i t h A R D S [ 1 0 ] . H o w e v e r , t h e s t u d y s h o w e d t h a t u s e o f c o r t i c o s t e r o i d s a t b a s e l i n e w a s h i g h e r t h a n 5 0 % a n d p o s t -All rights reserved. No reuse allowed without permission. author/funder, who has granted medRxiv a license to display the preprint in perpetuity. h o c a n a l y s e s r e v e a l e d a s t a t i s t i c a l l y s i g n i f i c a n t i n t e r a c t i o n b e t w e e n b a s e l i n e author/funder, who has granted medRxiv a license to display the preprint in perpetuity. All rights reserved. No reuse allowed without permission. author/funder, who has granted medRxiv a license to display the preprint in perpetuity. The copyright holder for this preprint (which was not peer-reviewed) is the . https://doi.org/10.1101/2020.04.01.20049700 doi: medRxiv preprint author/funder, who has granted medRxiv a license to display the preprint in perpetuity. The copyright holder for this preprint (which was not peer-reviewed) is the . https://doi.org/10.1101/2020.04.01.20049700 doi: medRxiv preprint I g a s a n e g a t i v e c o n t r o l . T h e s e c o n d s t a g e a n t i b o d y w a s F I T C a n t i - All rights reserved. No reuse allowed without permission. author/funder, who has granted medRxiv a license to display the preprint in perpetuity. R e s u l t s All rights reserved. No reuse allowed without permission. author/funder, who has granted medRxiv a license to display the preprint in perpetuity. The copyright holder for this preprint (which was not peer-reviewed) is the . https://doi.org/10.1101/2020.04.01.20049700 doi: medRxiv preprint s h o c k 3 3 % , a n t i -i n f l a m m a t o r y 1 7 % , A R D S 1 4 % a n d o t h e r 3 6 % ( f i b r o s i s , i m m u n o s u p p r e s s i o n p o s All rights reserved. No reuse allowed without permission. author/funder, who has granted medRxiv a license to display the preprint in perpetuity. All rights reserved. No reuse allowed without permission. author/funder, who has granted medRxiv a license to display the preprint in perpetuity. The copyright holder for this preprint (which was not peer-reviewed) is the . https://doi.org/10.1101/2020.04.01.20049700 doi: medRxiv preprint T o f u r t h e r c o n f i r m t h e s e f i n d i n g s , w e i s o l a t e d t h e n u c l e a r a n d c y t o p l a s m i c c o m p a r t m e n t s o f author/funder, who has granted medRxiv a license to display the preprint in perpetuity. author/funder, who has granted medRxiv a license to display the preprint in perpetuity. All rights reserved. No reuse allowed without permission. author/funder, who has granted medRxiv a license to display the preprint in perpetuity. The copyright holder for this preprint (which was not peer-reviewed) is the . https://doi.org/10.1101/2020.04.01.20049700 doi: medRxiv preprint A s S T A T 1 -S T A T 2 -I R F 9 c o m p l e x a l s o d r i v e s t h e e x p r e s s i o n o f H I F -1 a [ 3 2 ] , i t c a n b e e n v i s i o n e d t h a t a l s o t h e e x p r e s s i o n o f H I F -1 a i s d e c r e a s e d d u e t o g l u c o c o r t i c o i d u s e . I n a d d i t i o n , H I F -1 a m e d i a t e d t r a n s a c t i v a t i o n o f g e n e s m a y a l s o b e i m p a c t e d a s g l u c o c o r t author/funder, who has granted medRxiv a license to display the preprint in perpetuity. author/funder, who has granted medRxiv a license to display the preprint in perpetuity. The copyright holder for this preprint (which was not peer-reviewed) is the . https://doi.org/10.1101/2020.04.01.20049700 doi: medRxiv preprint T a b l e 1 . B a s e l i n e d e m o g r a p h i c s a n d 2 8 -d a y m o r t a l i t y i n t h e I F N b e t a -1 a a r m o f t h e I N T E author/funder, who has granted medRxiv a license to display the preprint in perpetuity. T a b l e 2 . B a s e l i n e d e m o g r a p h i c s a n d 2 8 -d a y m o r t a l i t y f o r p r o p e n s i t y -m a t c h e d I F N b e t a -1 a t r e a t e All rights reserved. No reuse allowed without permission. author/funder, who has granted medRxiv a license to display the preprint in perpetuity. T a b l e 3 . ( A ) I n d e p e n d e n t f a c t o r s a s s o c i a t e d w i t h 2 8 -d a y m o r t a l i t y i n a l l I F N b e t a -1 a t r e a t e All rights reserved. No reuse allowed without permission. author/funder, who has granted medRxiv a license to display the preprint in perpetuity. The copyright holder for this preprint (which was not peer-reviewed) is the All rights reserved. No reuse allowed without permission. author/funder, who has granted medRxiv a license to display the preprint in perpetuity. The copyright holder for this preprint (which was not peer-reviewed) is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. The copyright holder for this preprint (which was not peer-reviewed) is the . https://doi.org/10.1101/2020.04.01.20049700 doi: medRxiv preprint r e p r e s e n t t h e p l a c e b o i n d u c e d l e v e l o f C D 7 3 e x p r e s s i o n . S t a t i s t i c a l s i g n i f i c a n c e w a s a n a l y z e d u s i n All rights reserved. No reuse allowed without permission. author/funder, who has granted medRxiv a license to display the preprint in perpetuity. The copyright holder for this preprint (which was not peer-reviewed) is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. 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