Document text
Principal Investigator: Benjamin L King
Organization: UNIVERSITY OF MAINE ORONO
Fiscal Year: 2023
Award: $261,916
Funding agency: National Institute of General Medical Sciences
PROJECT SUMMARY / ABSTRACT
Skeletal muscle myopathy has been reported following infection by influenza virus. In Influenza-Associated
Myopathy (IAM), a systemic influenza infection can result in acute skeletal muscle damage that ranges from non-
specific degeneration to extensive necrosis. IAM is associated with high serum creatine kinase (CK) levels.
During the 2009 H1N1 influenza epidemic, 62% of hospitalized patients had increased serum CK levels. Given
that an estimated 9-45 million individuals acquire influenza infections annually in the US, developing new
strategies to reduce muscle damage are needed as influenza vaccines are difficult to design because of
unpredictable changes in viral strains within and across populations. The severity of viral disease varies between
individuals and depends on how the immune system responds to infection. One roadblock to understanding the
pathogenesis of IAM is that the relative contributions of the virus and host factors in vivo are not well understood.
Biopsy studies cannot show the temporal dynamics of viral invasion and subsequent recruitment of neutrophils
and macrophages into muscle. The zebrafish is a powerful model to study host-pathogen interactions as genetic
tools can be combined with in vivo imaging of transparent embryos. My laboratory uses a recently-developed
zebrafish model of influenza A virus (IAV) infection where it was shown that: 1) IAV-infected zebrafish embryos
exhibited mild muscle degeneration with sarcolemma damage and compromised extracellular matrix (ECM)
adhesion; and 2) neutrophils localize to sites of muscle damage in IAV-infected embryos. Our specific goal in
this proposal is to determine the mechanisms through which neutrophils influence the pathology of IAM. This
project will test the novel hypothesis that overactivation of neutrophils during IAV infection triggers a damaging
hyperinflammatory response that contributes to myopathy. In the first aim, we will test the hypothesis that
reduction in reactive oxidative species (ROS) production following IAV infection will limit damage by
strengthening muscle cell-ECM adhesion, and increase survival. This will be accomplished by examining how
global ROS reduction and neutrophil-specific ROS reduction alters muscle degeneration, cell-ECM adhesion,
and neutrophil localization in the muscle in vivo using IAV multi-spectral fluorescent reporter (Color-flu) strains.
In the second aim, we will test the hypothesis that defects in neutrophil migration following IAV infection will
increase muscle degeneration, and weaken ECM adhesion. To accomplish this, we will use in vivo confocal
imaging to study two zebrafish mutants with defective neutrophil migration infected with Color-flu to test our
hypotheses that: 1) neutrophil invasion into skeletal muscle is reduced, and 2) muscle degeneration is increased
and cell-ECM adhesion is increased over controls. One of these mutants overexpresses miR-199 in neutrophils
that disrupts neutrophil migration. Proposed RNA sequencing will allow us to determine microRNA gene
regulatory networks thereby allowing us to establish a link between IAM and microRNA genetic regulation.
Knowledge gained through the study of IAM may inform studies of other viral-associated myopathies.
Terms: <ADP Phosphocreatine Phosphotransferase><ATP Creatine Phosphotransferase><Acetylcysteine><Acetylin><Acute><Adhesions><Airbron><Antioxidants><Arrowhead><Biopsy><Biosensor><Blood Neutrophil><Blood Polymorphonuclear Neutrophil><Blood Serum><Body Tissues><Brachydanio rerio><Broncholysin><Brunac><Cell Communication and Signaling><Cell Locomotion><Cell Migration><Cell Movement><Cell Signaling><Cell-Extracellular Matrix><Cellular Migration><Cellular Motility><Cellular Regulation><Chemotaxis><Creatine Kinase><Creatine Kinase-B><Creatine Kinase-B Chain><Creatine Phosphokinase><Cues><Danio rerio><Defect><Disease><Disorder><ECM><Embryo><Embryonic><Exhibits><Extensive Necrosis><Extracellular Matrix><Fabrol><Fiber><Flu epidemic><Fluatox><Fluimucetin><Fluimucil><Fluprowit><Focal Infection><Genetic><Glutathione><Goals><Grant><Grippe><H1N1><H1N1 Virus><H2O2><Hemi-Myeloperoxidase><Hospital Admission><Hospitalization><Host Factor><Host Factor Protein><Hour><Hydrogen Peroxide><Hydroperoxide><Immune system><Individual><Infection><Inflammatory Response><Influenza><Influenza A><Influenza A Virus, H1N1 Subtype><Influenza A virus><Influenza Vaccines><Influenza Virus><Influenza Viruses Type A><Influenzavirus A><Innate Immune System><Innate Immunity><Integration Host Factors><Intracellular Communication and Signaling><Invaded><Knowledge><Laboratories><Ligands><Link><Macrophage><Marrow Neutrophil><Mediating><Mercapturic Acid><Micro RNA><MicroRNAs><Modeling><Motility><Muco Sanigen><Mucocedyl><Mucolator><Mucolyticum><Mucomyst><Mucosolvin><Mucret><Muscle><Muscle Cells><Muscle Disease><Muscle Disorders><Muscle Tissue><Muscular Diseases><Myelogenous><Myeloid><Myeloperoxidase><Myocarditis><Myocytes><Myopathic Conditions><Myopathic Diseases and Syndromes><Myopathic disease or syndrome><Myopathy><Mφ><N-Acetylcysteine><NAC Zambon><Native Immunity><Natural Immunity><Neo-Fluimucil><Neutrophil Activation><Neutrophil Infiltration><Neutrophil Recruitment><Neutrophilic Granulocyte><Neutrophilic Infiltrate><Neutrophilic Leukocyte><Non-Specific Immunity><Nonspecific Immunity><Orthomyxovirus Type A><Oxidative Stress><Parvolex><Pathogenesis><Pathology><Pathway interactions><Patients><Peroxidases><Polymorphonuclear Cell><Polymorphonuclear Leukocytes><Polymorphonuclear Neutrophils><Population><Production><RNA Seq><RNA sequencing><RNAseq><Regulation><Reporter><Reporting><Respaire><Respiratory Signs and Symptoms><Role><Sagittaria><Sarcolemma><Serum><Severities><Signal Pathway><Signal Transduction><Signal Transduction Systems><Signaling><Site><Skeletal Muscle><Symptoms><System><Testing><Tissues><Tixair><Transgenes><Type A Influenza><Viral><Viral Diseases><Virus><Virus Diseases><Visualization><Voluntary Muscle><Zebra Danio><Zebra Fish><Zebrafish><airway symptom><anti-oxidant><biological adaptation to stress><biological sensor><biological signal transduction><cardiac inflammation><cell behavior><cell growth regulation><cell motility><cellular behavior><confocal imaging><design><designing><develop therapy><extracellular><flu infection><flu vaccine><flu virus infection><flu virus vaccine><gamma-L-Glu-L-Cys-Gly><gamma-L-Glutamyl-L-Cysteinylglycine><gene regulatory network><imaging in vivo><in vivo><in vivo imaging><infected with flu><infected with flu virus><infected with influenza><infected with influenza virus><infection localized><influenza epidemic><influenza infection><influenza virus infection><influenza virus vaccine><influenzavirus><innate immune function><insight><intervention development><local infection><miRNA><miRNAs><migration><muscle degeneration><muscle strengthening><muscular><muscular disorder><mutant><neutrophil><novel><overexpress><overexpression><pathogen><pathway><reaction; crisis><recruit><respiratory symptom><response><social role><strength training><stress response><stress; reaction><therapy development><tool><transcriptome sequencing><transcriptomic sequencing><transgene><treatment development><vaccine against flu><vaccine against influenza><viral infection><virus infection><virus-induced disease>