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Principal Investigator: Nicholas S Heaton
Organization: DUKE UNIVERSITY
Fiscal Year: 2024
Award: $713,913
Funding agency: National Institute of Allergy and Infectious Diseases
PROJECT SUMMARY/ABSTRACT
Influenza viruses induce significant morbidity and mortality in the human population every year. Despite the
medical importance of influenza disease, the mechanisms underlying the viral pathogenesis are incompletely
understood. After influenza viruses initiate infection, cellular pattern recognition receptors detect components of
the virus and inflammatory cytokines such as type I interferon (IFN) are produced. While an important part of
early control of viral replication and spread, excessive or prolonged IFN signaling is associated with
immunopathology and more severe viral disease. We have previously shown that some respiratory epithelial
cells can survive direct viral infection, display prolonged IFN signaling, and contribute to lung immunopathology
likely by delaying epithelial regeneration. Thus, understanding how type I IFN is downregulated represents a
critical aspect of influenza disease pathogenesis. Interestingly, however, we failed to detect a difference in any
of the canonical IFN induction or suppression pathways in the cells that survived direct viral infection. We
therefore hypothesized that there must be additional regulators of IFN signaling relevant for IFN control
during the resolution of influenza virus-induced disease. By leveraging genome wide CRISPR activation
screening approaches, we were able to identify a previously uncharacterized regulator of IFN signaling (the ETS-
family transcription factor ETV7) which upon induction, non-uniformly suppressed the expression of individual
interferon stimulated genes (ISGs). The major goal of this application is to define this novel, ETV7 mediated, IFN
regulatory mechanism and understand its physiological importance during influenza viral disease in vivo. In Aim
1, we will perform high-throughput biochemical analysis of ETV7 binding across all ISG promoters and then
define how intrinsic ETV7 affinities for primary promoter sequences affect transcription factor competition. This
work will reveal the first regulatory pathway capable of differential suppression of individual ISGs. In Aim 2, we
will focus on understanding the cell type-specific importance of ETV7 regulation during respiratory epithelial
innate immune responses and tissue regeneration after infection. These experiments will establish a new
mechanism for how the balance between the antiviral signaling and regenerative capability in the lung is
maintained. Finally, in Aim 3, we will explore how dysregulation of ETS-family transcription factors affects viral
pathogenesis and recovery from infection in vivo. Successful completion of these studies will increase our
understanding of the mechanisms that regulate IFN signaling during influenza virus infection and potentially
inform new interventions to limit the severity of viral disease.
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disease rate><Murine><Mus><Native Immunity><Natural Immunity><Natural regeneration><Non-Specific Immunity><Nonspecific Immunity><Orthomyxovirus Type A><Orthomyxoviruses Type B><Pathogenesis><Pathway interactions><Pattern recognition receptor><Physiologic><Physiological><Play><Pneumonitis><Population><Position><Positioning Attribute><Production><Progenitor Cells><Proliferating><Promoter Regions><Promotor Regions><Pulmonary Inflammation><RNA Expression><Recovery><Regeneration><Regulation><Regulatory Element><Regulatory Pathway><Regulatory Protein><Research><Resolution><Respiratory Epithelium><Role><SARS corona virus 2><SARS-CO-V2><SARS-COVID-2><SARS-CoV-2><SARS-CoV2><SARS-associated corona virus 2><SARS-associated coronavirus 2><SARS-coronavirus-2><SARS-related corona virus 2><SARS-related coronavirus 2><SARSCoV2><Severe Acute Respiratory Coronavirus 2><Severe Acute Respiratory Distress Syndrome CoV 2><Severe Acute Respiratory Distress Syndrome Corona Virus 2><Severe Acute Respiratory Distress Syndrome Coronavirus 2><Severe Acute Respiratory Syndrome CoV 2><Severe Acute Respiratory Syndrome-associated coronavirus 2><Severe Acute Respiratory Syndrome-related coronavirus 2><Severe acute respiratory syndrome associated corona virus 2><Severe acute respiratory syndrome coronavirus 2><Severe acute respiratory syndrome related corona virus 2><Severities><Signal Pathway><Signal Repression><Signal Transduction><Signal Transduction Systems><Signaling><Site><Structure of respiratory epithelium><TEL Gene><Testing><Transcription><Transcription Factor Proto-Oncogene><Transcription factor genes><Transgenes><Type A Influenza><Variant><Variation><Viral><Viral Activity><Viral Diseases><Viral Function><Viral Pathogenesis><Viral Physiology><Virus><Virus Diseases><Virus Replication><Work><Wuhan coronavirus><activating CRISPR technology><airway epithelium><balance><balance function><biological signal transduction><cell culture><cell cultures><cell type><computational modeling><computational models><computer based models><computerized modeling><coronavirus disease 2019 virus><coronavirus disease-19 virus><cytokine><epithelium regeneration><experiment><experimental research><experimental study><experiments><flu infection><flu virus infection><gene modification><genetic promoter element><genetic promoter sequence><genetic regulatory protein><genetically modified><genome scale><genome-wide><genomewide><hCoV19><immunopathology><in vivo><infected with flu><infected with flu virus><infected with influenza><infected with influenza virus><influenza infection><influenza virus infection><influenzavirus><insight><interventional strategy><lung regeneration><model of animal><mortality><nCoV2><novel><overexpress><overexpression><pathogenic virus><pathway><preservation><prevent><preventing><progenitor><promoter><promoter sequence><promotor><prototype><pulmonary><pulmonary regeneration><regenerate><regenerate epithelium><regenerate new tissue><regenerate tissue><regenerating damaged tissue><regenerating tissue><regeneration model><regeneration potential><regenerative><regenerative model><regenerative potential><regulatory gene product><resolutions><respiratory tract epithelium><response><screening><screenings><social role><stem cells><tissue regeneration><tissue regrowth><tissue renewal><tissue specific regeneration><transcription factor><transgene><viral infection><viral multiplication><viral pathogen><viral replication><virus infection><virus multiplication><virus pathogen><virus pathogenesis><virus-induced disease>