Defective RNA Polymerase III sensing of mitochondrial DNA in pulmonary epithelial cells impairs type I IFN immunity to SARS-CoV-2

M Michelle Møhlenberg (Department of Biomedicine, Aarhus University) S Sofie Eg Jørgensen (Department of Biomedicine, Aarhus University) R Renée Marije van der Sluis (Department of Biomedicine, Aarhus University) T Thomas Zillinger (Department of Biomedicine, Aarhus University) D Daniëla Maria Hinke (Department of Biomedicine, Aarhus University) A Anne Kruse Hollensen (Department of Biomedicine, Aarhus University) A Anne Louise Hansen (Department of Biomedicine, Aarhus University) P Pierre Hausfater (Pitie Salpetriere Hospital, Departement de Sante Publique and Sorbonne University, INSERM, Institut Pierre Louis d‘Epidemiologie et de Sante Publique) G Guy Gorochov (Pitie Salpetriere Hospital, Departement de Sante Publique and Sorbonne University, INSERM, Institut Pierre Louis d‘Epidemiologie et de Sante Publique) F Florence Tubach (Pitie Salpetriere Hospital, Departement de Sante Publique and Sorbonne University, INSERM, Institut Pierre Louis d‘Epidemiologie et de Sante Publique) J Jade Ghosn (Assistance Publique–Hôpitaux de Paris.Nord, Hopital Bichat-Claude Bernard, Service des Maladies Infectieuses, Paris F75018, France and Universite Paris Cite, INSERM UMR 1137 IAME) C Cedric Laouenan (Assistance Publique–Hôpitaux de Paris.Nord, Hopital Bichat-Claude Bernard, Service des Maladies Infectieuses, Paris F75018, France and Universite Paris Cite, INSERM UMR 1137 IAME) M Merete Storgaard (Department of Infectious Diseases, Aarhus University Hospital) C Christian Kanstrup Holm (Department of Biomedicine, Aarhus University) H Helen Su (National Institute of Allergy and Infectious Diseases, NIH) R Rebeca Pérez de Diego (Institute of Biomedical Research of IdiPAZ, University Hospital “La Paz”) A Aurora Pujol (Neurometabolic Diseases Laboratory, Bellvitge Biomedical Research Institute, L’Hospitalet de Llobregat) S Shen-Ying Zhang (St. Giles Laboratory of Human Genetics of Infectious Diseases, The Rockefeller University) Q Qian Zhang F Fernanda Sales Luiz Vianna (Genomic Medicine, Experimental Research Center, Hospital de Clínicas de Porto Alegre) L Laurent Abel (St. Giles Laboratory of Human Genetics of Infectious Diseases, The Rockefeller University) A Aurélie Cobat (St Giles Laboratory of Human Genetics of Infectious Diseases, Rockefeller Branch, The Rockefeller University) J Jean-Laurent Casanova T Trine H. Mogensen (Department of Biomedicine, Aarhus University)

Abstract

The clinical spectrum of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection ranges from asymptomatic cases to critical COVID-19 pneumonia. To investigate the role of host genetics in susceptibility to critical COVID-19 and identify pathophysiological mechanisms and pathways, we analyzed whole-exome and whole-genome sequencing data from the COVID Human Genetic Effort. We identified 10 rare, monoallelic predicted loss-of-function variants in 18 patients in POLR3A and POLR3C encoding two subunits of RNA polymerase III (POL III), a nuclear multisubunit enzyme, which has been implicated in cytosolic DNA sensing. These variants were deleterious for expression of full-length POLR3A and POLR3C proteins. We demonstrate that human pulmonary A549-hACE2 cells with reduced POLR3A or POLR3C expression exhibit impaired type I IFN responses to transfected mitochondrial DNA (mtDNA) or SARS-CoV-2 infection, together with increased viral replication. Mechanistically, we show that SARS-CoV-2 induces cellular mtDNA release via oligomerization of the mitochondrial voltage-dependent anion channel under virus-induced oxidative stress, enabling POL III–mtDNA interaction. These findings establish POL III as a sensor of endogenous mtDNA released during viral infection and indicate that autosomal dominant POL III haploinsufficiency may predispose individuals to critical COVID-19.

Article Details

Volume / Issue Vol. 123, Issue 12
Published March 24, 2026
ISSN 0027-8424
Publisher National Academy of Sciences

Authors (24)

M

Michelle Møhlenberg

Department of Biomedicine, Aarhus University

S

Sofie Eg Jørgensen

Department of Biomedicine, Aarhus University

R

Renée Marije van der Sluis

Department of Biomedicine, Aarhus University

T

Thomas Zillinger

Department of Biomedicine, Aarhus University

D

Daniëla Maria Hinke

Department of Biomedicine, Aarhus University

A

Anne Kruse Hollensen

Department of Biomedicine, Aarhus University

A

Anne Louise Hansen

Department of Biomedicine, Aarhus University

P

Pierre Hausfater

Pitie Salpetriere Hospital, Departement de Sante Publique and Sorbonne University, INSERM, Institut Pierre Louis d‘Epidemiologie et de Sante Publique

G

Guy Gorochov

Pitie Salpetriere Hospital, Departement de Sante Publique and Sorbonne University, INSERM, Institut Pierre Louis d‘Epidemiologie et de Sante Publique

F

Florence Tubach

Pitie Salpetriere Hospital, Departement de Sante Publique and Sorbonne University, INSERM, Institut Pierre Louis d‘Epidemiologie et de Sante Publique

J

Jade Ghosn

Assistance Publique–Hôpitaux de Paris.Nord, Hopital Bichat-Claude Bernard, Service des Maladies Infectieuses, Paris F75018, France and Universite Paris Cite, INSERM UMR 1137 IAME

C

Cedric Laouenan

Assistance Publique–Hôpitaux de Paris.Nord, Hopital Bichat-Claude Bernard, Service des Maladies Infectieuses, Paris F75018, France and Universite Paris Cite, INSERM UMR 1137 IAME

M

Merete Storgaard

Department of Infectious Diseases, Aarhus University Hospital

C

Christian Kanstrup Holm

Department of Biomedicine, Aarhus University

H

Helen Su

National Institute of Allergy and Infectious Diseases, NIH

R

Rebeca Pérez de Diego

Institute of Biomedical Research of IdiPAZ, University Hospital “La Paz”

A

Aurora Pujol

Neurometabolic Diseases Laboratory, Bellvitge Biomedical Research Institute, L’Hospitalet de Llobregat

S

Shen-Ying Zhang

St. Giles Laboratory of Human Genetics of Infectious Diseases, The Rockefeller University

Q

Qian Zhang

F

Fernanda Sales Luiz Vianna

Genomic Medicine, Experimental Research Center, Hospital de Clínicas de Porto Alegre

L

Laurent Abel

St. Giles Laboratory of Human Genetics of Infectious Diseases, The Rockefeller University

A

Aurélie Cobat

St Giles Laboratory of Human Genetics of Infectious Diseases, Rockefeller Branch, The Rockefeller University

J

Jean-Laurent Casanova

T

Trine H. Mogensen

Department of Biomedicine, Aarhus University