National Jewish Health

GENETIC EPIDEMIOLOGY OF COPD

AIRWAY TRANSCRIPTOMIC RESPONSES TO COVID-19 ILLNESSES IN THE HUMAN EPIDEMIOLOGY AND RESPONSE TO SARS-COV-2 (HEROS) COHORT

(1 OF 2) GENETIC EPIDEMIOLOGY OF COPD

OVERALL ATOPIC DERMATITIS RESEARCH NETWORK

ETHER LIPIDS, EICOSANOIDS, AND LUNG CELL PATHOPHYSIOLOGY

LUNG GENOMICS RESEARCH CONSORTIUM

MECHANISMS AND TREATMENT OF DEPLOYMENT-RELATED LUNG INJURY: REPAIR OF THE INJURED EPITHELIUM

DEFINING MOLECULAR PHENOTYPES OF EXACERBATION PRONE ASTHMATICS

AN ASTHMA COLLABORATION TO REDUCE CHILDHOOD ASTHMA DISPARITIES ON THE NAVAJO NATION

CENTER FOR RESPIRATORY BIODEFENSE AT NATIONAL JEWISH MEDICAL AND RESEARCH CENTER

INFLAMMATION, AIRWAYS REACTIVITY AND ASTHMA

CERAMIDE-INDUCED LUNG DESTRUCTION IN EMPHYSEMA

ROLE OF KLF15 IN AIRWAY SMOOTH MUSCLE AND THE RESPONSE TO GLUCOCORTICOIDS

BLACK LUNG

LUNG MACROPHAGE PROGRAMMING IN ACUTE LUNG INJURY

COMMUNITY PROJECT FUNDING/CONGRESSIONALLY DIRECTED SPENDING - CONSTRUCTION

ANTI IL5 AND CHURG STRAUSS SYNDROME: A DOUBLE BLIND, PLACEBO CONTROLLED TRIAL

ENVIRONMENTAL DETERMINANTS OF AIRWAY DISEASE IN CHILDREN

CHARACTERISTICS OF T CELL RECEPTORS

ANTIGEN RECOGNITION BY LYMPHOCYTES

ASTHMA SUSCEPTIBILITY DUE TO ENVIRONMENTAL PROGRAMING OF INNATE IMMUNITY IN UTERO

CLINICAL CENTERS FOR THE NHLBI ASTHMA NETWORK (ASTHMANET)

USE OF THE SRC FAMILY KINASE INHIBITOR SARACATINIB IN THE TREATMENT OF PULMONARY FIBROSIS

MESENCHYMAL VASCULAR PROGENITOR DEPLETION PROMOTES LUNG AGING AND SUSCEPTIBILITY TO EMPHYSEMA - THE OVERALL MISSION OF THIS RESEARCH PROGRAM IS TO DEFINE HOW PULMONARY MESENCHYMAL VASCULAR PROGENITOR DEPLETION PROMOTES LUNG AGING AND SUSCEPTIBILITY TO EMPHYSEMA. LOSS OF EPITHELIAL PROGENITOR CELL FUNCTION IS A UNIFYING FACTOR IN ACCELERATED LUNG AGING, AND THE DEVELOPMENT OF EMPHYSEMA. HOWEVER, EQUALLY AS IMPORTANT BUT POORLY UNDERSTOOD, THERE IS A GAP IN OUR UNDERSTANDING OF MESENCHYMAL VASCULAR PROGENITORS (MVPC) IN THESE PROCESSES. THIS PROPOSAL IS SIGNIFICANT AS IT ATTEMPTS TO FILL IN A NUMBER OF IMPORTANT GAPS SURROUNDING HOW DYSFUNCTION OF THE MVPC PROGENITOR POPULATION, CONTRIBUTES TO AGING AND INCREASED SUSCEPTIBILITY TO EMPHYSEMA VIA REGULATION OF VASCULAR REMODELING AND LOSS OF ANGIOSTASIS. ANALYSIS OF THE MVPC - LUNG NICHE INTERACTIONS PROVIDE A TARGET RICH ENVIRONMENT TO IDENTIFY NUANCES IN MVPC PROGENITOR DEPENDENT PATHWAYS RELEVANT TO THE PATHOBIOLOGY OF ANGIOSTASIS, AGING AND EMPHYSEMA, AS WELL AS THE POTENTIAL TO IDENTIFY THERAPEUTICS TO RESTORE TISSUE FUNCTION. WE PROPOSE THREE FOCUS AREAS FOR OUR RESEARCH BASED ON COMPLEMENTARY THEMES. THEME 1 REGULATION OF MVPC FUNCTION IN HEALTHY AND AGED LUNG: WE WILL USE OUR UNIQUE IN VIVO AND IN VITRO MODEL SYSTEMS TO IDENTIFY HOW MVPC FUNCTION AND ADAPTIVE ANGIOGENESIS IS REGULATED DURING TISSUE HOMEOSTASIS AND AGING. THEME 2 CONSEQUENCE OF MVPC LOSS OF FUNCTION IN HEALTHY, AGED AND CIGARETTE SMOKE EXPOSED LUNG: WE WILL SHOW THAT LOSS OF MVPC FUNCTION, BY DEPLETION OR ALTERED SIGNALING, DRIVES VASCULOPATHY AND SUBSEQUENT LUNG AGING AND EMPHYSEMA. THEME 3 THERAPEUTIC RESCUE OF MVPC FUNCTION IN AGED AND CIGARETTE SMOKE EXPOSED LUNG: WE WILL VALIDATE THE USE OF MVPC AND REPURPOSING OF FDA APPROVED PAQUINIMOD TO RESTORE MVPC NUMBERS AND FUNCTION, SUBSEQUENT TISSUE FUNCTION AND ESTABLISH TIME FRAMES FOR INTERVENTION. POSITIVE RESULTS FROM THESE STUDIES ARE READILY TRANSLATABLE. WE WILL LEVERAGE OUR STRONG COLLABORATIONS, AT NATIONAL JEWISH, UNIVERSITY OF COLORADO AS WELL AS INTERNATIONALLY RECOGNIZED COLLABORATORS PROVIDE A BASIC AND TRANSLATIONAL UNDERSTANDING OF THE MECHANISMS REGULATING MVPC FUNCTION AND DIFFERENTIATION AT THE SINGLE CELL LEVEL AS WELL AS HOW THEY REGULATE THEIR NICHE AND LUNG MICROENVIRONMENT. WE WILL ALSO DEFINE WHETHER PROGENITOR RESCUE WITH MVPC CELL THERAPY OR INTERVENTIONAL TREATMENT WILL RESTORE LUNG STRUCTURE AND FUNCTION.

METABOLIC PROFILES OF COPD PHENOTYPES

MOLECULAR MECHANISMS OF IMMUNE TOLERANCE

TRANSCRIPTOMIC AND PHARMACOGENETIC ASTHMA ENDOTYPES IN MINORITY CHILDREN

A PROSPECTIVE STUDY OF T CELL IMMUNE-PHENOTYPE AND DISEASE OUTCOME IN PULMONARY SARCOIDOSIS

MECHANISMS OF RIGHT VENTRICLE ADAPTATION TO PULMONARY HYPERTENSION

ROLE OF LUNG MSC IN EMPHYSEMA

OPTIMIZATION OF AEOL10150 TREATMENT OF SULFUR MUSTARD-INDUCED LUNG TOXIDROME IN A PIG MODEL

THE ROLE OF BACTERIAL TOXINS IN HUMAN SKIN DISEASE.

STEPPED-CARE MANAGEMENT OF INSOMNIA CO-OCCURRING WITH SLEEP APNEA

PARKIN IN MITOCHONDRIAL DYSFUNCTION AND AIRWAY INFLAMMATION OF OBESE ASTHMA

APOPTOSIS AND DEFECTIVE REPAIR IN COPD

BIOMARKER OF LUNG DISEASE IN AFRICAN AMERICANS

BLACK LUNG

TAILORED TREATMENT TO ENHANCE RISK PERCEPTION IN SLEEP APNEA

ROLE OF FABP5 IN COPD EXACERBATIONS

GENETIC RISK FOR GRANULOMATOUS INTERSTITIAL LUNG DISEASE

USING MULTI-OMICS TO DEFINE REGULATORS AND DRIVERS OF GRANULOMATOUS INFLAMMATION AND CHRONIC BERYLLIUM DISEASE - PROJECT SUMMARY/ABSTRACT: THE GOAL OF THIS STUDY IS TO DEFINE CELL-SPECIFIC REGULATORY NETWORKS AND KEY DRIVERS OF CELLULAR RESPONSE TO BERYLLIUM (BE) THAT RESULT IN THE GRANULOMATOUS LUNG DISEASE, CHRONIC BERYLLIUM DISEASE (CBD), AT THE SITE OF ORGAN INVOLVEMENT. RELYING ON THE COMPLEMENTARY, UNIQUE EXPERTISE OF INVESTIGATIVE TEAM WE WILL DEFINE PATHOGENIC PATHWAYS AND RISK FACTORS FOR CBD, ITS PRECURSOR (BE SENSITIZATION; BES) AND SIMILAR ENVIRONMENTALLY INDUCED DISEASES. EXPOSURE TO AN INHALED BE ANTIGEN(S), IN THE SETTING OF A GENETICALLY SUSCEPTIBLE HOST, INITIATES A TH1 IMMUNE RESPONSE, WITH ANTIGEN PRESENTATION OCCURRING VIA HLA CLASS II ON ANTIGEN PRESENTING CELL (APC) IN THE CONTEXT OF CD4+ T CELLS. SUBSEQUENTLY, CD4+ T CELLS AND APCS ARE RECRUITED TO THE LUNG, PROLIFERATE, PRODUCE CYTOKINES AND CHEMOKINES, AND EVENTUALLY FORM GRANULOMAS. AN INCREASED PREVALENCE OF HLA-DPB1 ALLELES WITH A GLUTAMIC ACID AT AMINO ACID POSITION 69 (E69) IS FOUND IN CBD AND BES, ALTHOUGH THIS VARIANT IS FOUND IN UP TO 40% OF BE EXPOSED WORKERS WITHOUT BES OR CBD, SUGGESTING THAT OTHER FACTORS OR FORMS OF GENETIC REGULATION ARE IMPORTANT IN DISEASE PATHOGENESIS. GROWING DATA IN OTHER IMMUNE-MEDIATED DISEASES SUGGESTS THAT EPIGENETIC MECHANISMS IN COMBINATION WITH GENETIC SUSCEPTIBILITY AND ENVIRONMENT MAY HELP EXPLAIN DISEASE RISK. EPIGENETIC MODIFICATIONS DETERMINE T CELL AND APC/MACROPHAGE DIFFERENTIATION AND THE ENSUING IMMUNE RESPONSE THROUGH DNA METHYLATION AND HISTONE MODIFICATIONS OF KEY GENES AND THUS IMPACT HEALTH AND DISEASE. OUR PREVIOUS WORK DEMONSTRATED DNA METHYLATION CHANGES ASSOCIATED WITH ALTERATIONS IN GENE EXPRESSION AND DISEASE STATE IN BRONCHOALVEOLAR LAVAGE (BAL) CELLS OF CBD SUBJECTS COMPARED TO BES AND CONTROLS, INCLUDING PIVOTAL IMMUNE RESPONSE GENES AND NETWORKS. SINGLE CELL SEQUENCING TECHNOLOGIES HAVE EMERGED AS A KEY METHOD TO CHARACTERIZE NOVEL CELL POPULATION AND CELL-SPECIFIC CHANGES IN GENE EXPRESSION. BASED ON THIS INFORMATION, OUR HYPOTHESIS IS THAT EXPOSURE TO BE ALTERS EPIGENETIC MARKS, IMPACTING GENE EXPRESSION AND IMMUNE CELL DIFFERENTIATION IN CELL-SPECIFIC MANNER, AND ULTIMATELY RISK OF GRANULOMATOUS LUNG DISEASE. WE WILL TEST THIS HYPOTHESIS THROUGH THREE SPECIFIC AIMS, FOCUSING MAINLY ON MACROPHAGES AND CD4+ T CELLS BUT ACKNOWLEDGING THAT OTHER CELL POPULATIONS ARE IMPORTANT AND CONSIDERING THEM IN ALTERNATIVE APPROACHES AND FUTURE DIRECTIONS. IN AIM 1 WE WILL CHARACTERIZE THE LUNG IMMUNE CELL RESPONSE TO BE EXPOSURE USING SINGLE CELL ATAC-SEQ AND CITE- SEQ TO MEASURE CHROMATIN ACCESSIBILITY, GENE EXPRESSION LEVELS AND PROTEIN EPITOPES AT FOUR TIMEPOINTS TO DEFINE KEY DRIVERS IN SPECIFIC CELL POPULATIONS. IN AIM 2 WE WILL VALIDATE THE REGULATORY NETWORKS IN UNCULTURED LUNG SAMPLES, FOCUSING ON MACROPHAGES AND CD4+ TCELLS, USING BULK ATAC- ME AND RNA-SEQUENCING. WE WILL FUNCTIONALLY VALIDATE THE RESULTS FROM OUR MULTI-OMIC ANALYSIS FROM AIMS 1 AND 2 USING CRSIPR-DCAS9 TO CHANGE METHYLATION, CHROMATIN ACCESSIBILITY, AND GENE EXPRESSION AT LOCI IDENTIFIED AS KEY DRIVERS OF RESPONSE TO BE IN CULTURED MACROPHAGE AND T CELL LINES IN AIM 3. ULTIMATELY, THIS PROPOSAL WILL ENHANCE OUR UNDERSTANDING OF THE NOVEL GENES, REGULATORY PATHWAYS AND NETWORKS, AND MOLECULAR MECHANISMS INVOLVED IN CBD ETIOLOGY.

REGULATION OF GENE EXPRESSION IN THE ANAPHYLACTIC PATHWAY

NEUTROPHIL NOX2 CONTROLS MONONUCLEAR CELL FUNCTIONS IN INFLAMMATION; ROLE IN CGD

GENETIC CONTROL OF AIRWAY EPITHELIUM GENE EXPRESSION IN CHILDHOOD ASTHMATICS

MACROPHAGE APOPTOSIS IN RESOLUTION OF ACUTE LUNG INJURY

IDENTIFICATION OF PATHWAYS TO MITIGATE IMMUNE-RELATED ADVERSE EVENTS WITH CANCER IMMUNOTHERAPY

ILC2 MEMORY IN ASTHMA

ROLES FOR INTERSTITIAL AND AIRSPACE MACROPHAGES IN RESOLUTION OF PULMONARY INFLAMMATION

THE BREATHEWELL PROGRAM TO IMPROVE ASTHMA OUTCOMES

TELECOMMUNICATIONS ENHANCED ASTHMA MANAGEMENT (TEAM)

ROLE OF IMMUNOPROTEASOME IN AIRWAY VIRAL INFECTION

SUNBEAM BIRTH COHORT: DECIPHERING THE ROLE OF EARLY LIFE SKIN DYSFUNCTION IN DEVELOPMENT OF THE ALLERGIC MARCH - PROJECT SUMMARY/ABSTRACT ALLERGIC DISEASES, INCLUDING ATOPIC DERMATITIS (AD), FOOD ALLERGIES (FA), AND ASTHMA ARE COMMON IN YOUNG CHILDREN AND ARE OFTEN CO-MORBID, SUGGESTING A SHARED ALLERGIC PATHOBIOLOGY. SUPPORTING THIS, DEVELOPMENT OF THESE DISEASES OFTEN OCCURS IN A SEQUENTIAL PATTERN ACROSS INFANCY KNOWN AS THE “ALLERGIC MARCH,” WITH A DIAGNOSIS OF AD BEFORE 12-MONTHS RESULTING IN INCREASED RISK OF FA AND ASTHMA. HOWEVER, ONLY A SUBSET OF INFANTS WITH AD PROCEED DOWN THE ALLERGIC MARCH FOR UNKNOWN REASONS. WE HYPOTHESIZE THAT DIFFERENT PATHOBIOLOGICAL MECHANISMS (I.E. ENDOTYPES) OPERATING IN THE SKIN, UNDERLIE AD CLINICAL SUBTYPES AND THEIR PROPENSITY TO GIVE RISE TO ADDITIONAL ALLERGIC DISEASES. SUPPORTING THIS, WE HAVE FOUND A TYPE 2 INFLAMMATORY SKIN ENDOTYPE THAT WAS GREATLY INCREASED IN AD SUBJECTS WITH FA RELATIVE TO AD SUBJECTS WITHOUT FA, WITH THE ADFA+ PATIENTS ALSO EXHIBITING SKIN BARRIER ABNORMALITIES. MOREOVER, ANIMAL MODELS HAVE SHOWN THAT SKIN BARRIER DISRUPTION WITH INFLAMMATION CAN DRIVE SENSITIZATION TO ALLERGENS AND THE DEVELOPMENT OF AD, FA, AND AIRWAY INFLAMMATION. GENETIC STUDIES ALSO SUPPORT A SHARED GENETIC DISPOSITION AMONG ALLERGIC DISEASES AND IMPLICATE DYSREGULATION OF SKIN GENES IN THIS RISK. AMONG ADULTS, HETEROGENEITY IN INFLAMMATORY SKIN ENDOTYPE HAS BEEN OBSERVED, INCLUDING TH22, TH17, IL36G, AND TH1 PATHWAY ACTIVATION. THE ROLES OF THESE ENDOTYPES IN PROGRESSION OF INFANTS THROUGH THE ALLERGIC MARCH IS UNEXPLORED. WE HAVE PIONEERED SKIN TAPE STRIPPING (STS) RNA-SEQ METHODS TO ALLOW REPEATED, MINIMALLY INVASIVE SKIN EXPRESSION PROFILING. USING THESE METHODS, WE WILL EVALUATE WHETHER EARLY-LIFE SKIN PATHOBIOLOGY LEADS TO THE DEVELOPMENT OF THE ALLERGIC MARCH AND OTHER SEQUENCES OF ALLERGIC DISEASE DEVELOPMENT. OUR PROPOSAL RELIES ON PHENOTYPING AND STS SAMPLES COLLECTED IN THE SYSTEMS BIOLOGY OF EARLY ATOPY (SUNBEAM) BIRTH COHORT (N=2500). FROM A SUNBEAM ALLERGIC DISEASE CASE-COHORT (N~800) WE WILL GENERATE RNA-SEQ DATA ON STS SAMPLES COLLECTED AT BIRTH, 2, 5, 12, 24, AND 36 MONTHS, PAIRED WITH ALLERGIC DISEASE PHENOTYPING. IN AIM 1 WE WILL DETERMINE INFLAMMATORY ENDOTYPES OF AD IN INFANCY AND THEIR ASSOCIATED PATTERNS OF GENE EXPRESSION DYSREGULATION. CELLULAR DECONVOLUTION OF RNA-SEQ WILL IDENTIFY IMMUNE CELL TYPES AND CHANGES IN EPIDERMAL CELLULAR COMPOSITION THAT UNDERLIE DIFFERENT AD ENDOTYPES. WE WILL DETERMINE WHETHER SKIN EXPRESSION AND ENDOTYPES, AT BIRTH AND WITH AD AT 2 AND 5 MOS, ARE PREDICTIVE OF FOOD ALLERGY AT ONE YEAR OF AGE. IN AIM 2 WE WILL DETERMINE LONGITUDINAL PATTERNS OF EARLY LIFE SKIN PATHOBIOLOGY THAT UNDERLIE THE ALLERGIC MARCH AND OTHER SEQUENCES OF ALLERGIC DISEASE DEVELOPMENT. WE WILL DEFINE ALLERGIC DISEASE DEVELOPMENTAL CLASSES, THEIR ASSOCIATED LONGITUDINAL EXPRESSION AND CELLULAR COMPOSITION PROFILES, AND PROSPECTIVE BIOMARKERS FOR ALLERGIC DISEASE DEVELOPMENT. IN AIM 3, WHOLE GENOME- AND RNA-SEQ DATA WILL BE USED TO IDENTIFY EQTLS THAT INFLUENCE SKIN EXPRESSION AT EACH TIMEPOINT AND LONGITUDINAL SKIN DEVELOPMENT AMONG ALLERGIC DISEASE GROUPS. USING TWAS, GENETIC MODELS OF EARLY-LIFE SKIN GENE EXPRESSION WILL BE USED WITH GWAS DATA FOR ALLERGIC DISEASES TO IDENTIFY GENETIC VARIANTS INFLUENCING ALLERGIC DISEASE RISK THROUGH MODULATION OF EARLY-LIFE SKIN GENE EXPRESSION.

EPIGENETIC REGULATION OF IMMUNE PATHWAYS IN SARCOIDOSIS

DISCOVERY OF NOVEL MECHANISMS OF ACTION OF UBIQUITIN-LIKE PROTEINS IN CELLULAR STRESS PATHWAYS

THE ORIGIN AND ROLE OF PULMONARY ILC2 SUBSETS IN ANTI-HELMINTH IMMUNITY

PROTEOLYSIS IN THE PATHOGENESIS OF ARDS - PROJECT SUMMARY/ABSTRACT THE ACUTE RESPIRATORY DISTRESS SYNDROME (ARDS) IS THE SEVEREST FORM OF ACUTE LUNG INJURY (ALI), AND IS CHARACTERIZED BY INJURY TO THE ALVEOLAR-CAPILLARY UNIT AND COMPROMISED ALVEOLAR EPITHELIAL INTEGRITY, LEADING TO HIGH PERMEABILITY PULMONARY EDEMA AND NEUTROPHILIC ALVEOLAR INFLAMMATION. FAILURE TO REPAIR THE DAMAGED ALVEOLAR MEMBRANE LEADS TO SIGNIFICANT MORTALITY. STUDIES FROM SEVERAL LABORATORIES, INCLUDING OUR OWN, HAVE FOUND THAT INDUCTION OF ALI IN ANIMAL MODELS IS ASSOCIATED WITH INCREASED EXPRESSION OF MATRIX METALLOPROTEINASE-3 (MMP-3), A PROTEASE THAT HAS BEEN SHOWN TO DIRECTLY TARGET CELL-CELL JUNCTIONS AND TO DEGRADE BASEMENT MEMBRANES, ALTHOUGH IT WAS UNKNOWN AT WHAT STAGES OF ALI PATHOLOGY THIS PROCESS WAS MOST CRITICAL. OTHER LABORATORIES AND OURS HAVE ALSO FOUND THAT TRANSGENIC MICE LACKING MMP-3 ARE RESISTANT TO LUNG INJURY INDUCED BY MULTIPLE STIMULI, INDICATING THAT INHIBITORS OF MMP-3 HAVE POTENTIAL AS THERAPEUTIC AGENTS FOR ARDS, ALTHOUGH SUCH A THERAPEUTIC STRATEGY WOULD NEED TO BE HIGHLY SELECTIVE, AS OTHER MMPS HAVE BEEN SHOWN TO PLAY KEY ROLES IN REPAIR OF LUNG INJURY. IN THIS TRANSLATIONAL APPLICATION, WE PROPOSE EXPERIMENTS WHICH WILL BRIDGE THESE CRITICAL GAPS. IN AIM 1, WE WILL USE PHYSIOLOGICALLY RELEVANT CELL CULTURE AND ANIMAL MODELS OF ALI/ARDS INCLUDING NOVEL TRANSGENIC MICE TO DEFINE HOW MMP-3 AFFECTS THE RESOLUTION OF EPITHELIAL INJURY IN RESPONSE TO ACID ASPIRATION OR INFLUENZA INFECTION. IN AIM 2, WE WILL USE OUR DIRECTED MOLECULAR EVOLUTION PLATFORM TO CREATE VARIANTS OF TISSUE INHIBITOR OF METALLOPROTEINASE-1 (TIMP-1) WITH GREATLY INCREASED AFFINITY FOR MMP-3 AND DECREASED BINDING TO BENEFICIAL MMPS, AND WE WILL DEFINE THE THERAPEUTIC UTILITY OF THESE TIMP VARIANTS IN MOUSE MODELS OF ALI/ARDS. IN AIM 3, WE WILL INTERROGATE CLINICAL SAMPLES OF ARDS TO DETERMINE THE PRECISE STAGES OF HUMAN DISEASE PROGRESSION AT WHICH THERAPEUTIC INTERVENTION WOULD BE MOST BENEFICIAL. OUR RESEARCH PROPOSAL WILL DEFINE A NEWLY-DISCOVERED MECHANISM BY WHICH MMP-3 DRIVES ARDS PATHOLOGY THROUGH AN INTEGRATED RESEARCH PLAN THAT LINKS SINGLE CELL RNA SEQUENCING OF CELL POPULATIONS ISOLATED FROM ARDS PATIENT TISSUE, CELL CULTURE AND ANIMAL MODELS OF ARDS, AND ANALYSIS OF RELEVANT ARDS BIOMARKERS AND EX VIVO CULTURE OF PRECISION CUT HUMAN LUNG SLICES, AND WILL DEVELOP A NOVEL SELECTIVE INHIBITOR WITH IMMEDIATE THERAPEUTIC POTENTIAL. OUR MULTIDISCIPLINARY RESEARCH TEAM IS UNIQUELY SUITED TO ADDRESS THESE QUESTIONS. OUR WORK WILL ADDRESS CRITICAL MECHANISMS UNDERLYING ALI/ARDS, WILL USE THIS KNOWLEDGE TO DEVELOP PERSONALIZED STRATEGIES THAT WILL IDENTIFY PATIENTS AT HIGHEST RISK OF DEVELOPING ALI/ARDS, AND WILL CREATE THERAPEUTIC APPROACHES BASED ON THE PIVOTAL ROLE OF MMP-3 IN THE PATHOGENESIS OF EPITHELIAL INJURY LEADING TO ALI/ARDS.

LOSS OF PROGENITOR FUNCTION ACCELERATES LUNG AGING - AGING IS ASSOCIATED WITH LOSS OF LUNG STRUCTURE AND DECLINING FUNCTION. EMPHYSEMATOUS LOSS OF TISSUE STRUCTURE IS EXACERBATED BY VASCULOPATHY, WHICH INCREASES SUSCEPTIBILITY TO LUNG DISEASE, AND LIMITS SURVIVAL. WE HAVE PREVIOUSLY DEMONSTRATED THAT THE STRUCTURE AND FUNCTION OF THE LUNG MICROVASCULATURE IS REGULATED BY A SPECIALIZED MESENCHYMAL VASCULAR PROGENITOR CELL (MVPC). WE HAVE ALSO DEFINED DICKKOPF-RELATED PROTEIN 1 (DKK1) AS A REGULATOR OF THIS NICHE IN MURINE AND HUMAN MODEL. OUR PRELIMINARY DATA DEMONSTRATE THAT MVPC NUMBERS DECLINE WITH AGE IN WT MICE BY 1 YEAR, AND THAT, WHEN MVPC ARE DEPLETED IN YOUNG MICE, LUNG AGING IS ACCELERATED, RESULTING IN SEVERE EMPHYSEMA AT 1.5 YEARS OF AGE. WHILE MVPC ARE KEY MODULATORS OF THE PULMONARY MICROVASCULATURE IN THE DISTAL LUNG, ADVENTITIAL STEM CELLS (ASC) INFLUENCE LARGE BLOOD VESSEL HOMEOSTASIS IN THE PROXIMAL LUNG. OUR PRELIMINARY DATA IDENTIFIES KEY SIMILARITIES BETWEEN MVPCS AND ASCS SUGGESTING THAT DECLINE IN ASC NUMBERS AND FUNCTION MAY ALSO RESULT IN ACCELERATED LUNG AGING. GIVEN ASC FUNCTION IS TIGHTLY CONTROLLED BY LUNG-RESIDENT TYPE-2 INNATE LYMPHOID CELLS (ILC2) AND PULMONARY ILC2 ARE KNOWN TO DECREASE WITH AGE, LOSS OF ASC-ILC2 CROSSTALK IS LIKELY A CONTRIBUTOR TO EMPHYSEMA. SIMILAR TO AGING, VASCULAR REMODELING IS DRIVEN BY CHRONIC ACTIVATION OF TYPE 2 (TH2) INFLAMMATION. OUR PRELIMINARY DATA SUPPORT INCREASED TH2 CYTOKINE PROFILES IN PLASMA AND BAL OF COPD PATIENTS. HUMAN AND MURINE SCRNASEQ DATA DEFINE ILC2 AND MVPC POPULATIONS AS WELL AS IMMUNE CELL INFILTRATE AND CYTOKINE EXPRESSION CHARACTERISTIC OF TH2 INFLAMMATION IN COPD AND A SMOKING MODEL. THE GOAL OF THIS PROPOSAL IS TO DEFINE THE MECHANISMS THAT RESULT IN LOSS OF ADULT MVPC AND ILC2 PROGENITOR FUNCTION CONTRIBUTING TO ACCELERATED LUNG AGING, BY ALTERING VASCULAR STRUCTURE AND FUNCTION, AND INCREASED SUSCEPTIBILITY TO EMPHYSEMA IN THE AGED POPULATION. WE HYPOTHESIZE THAT LOSS OF PROGENITOR FUNCTION ACCELERATES LUNG AGING BY ALTERING CELL INTERACTIONS WITHIN VASCULAR NICHES, PROMOTING VASCULAR REMODELING AND INCREASING SUSCEPTIBILITY TO EMPHYSEMA IN THE AGING POPULATION. WE WILL TEST THAT DECLINE IN MVPC AND ILC2 NUMBERS AND/OR FUNCTION IN THE AGING LUNG ACCELERATES AGING VIA IMPAIRMENT OF VASCULAR HOMEOSTASIS DUE TO DISRUPTION OF CELL – CELL INTERACTIONS IN THEIR RESPECTIVE PERIVASCULAR NICHES, PROMOTING VASCULAR REMODELING AND LOSS OF TISSUE STRUCTURE USING NOVEL CONDITIONAL MURINE MODELS TO KNOCK DOWN PROGENITORS IN AGED MICE OR YOUNG MICE. WE WILL ASSESS THE REQUIREMENT OF MVPC, MVPC DERIVED DKK1, ILC2 CELLS, AND ILC2 DERIVED IL13 IN THE MAINTENANCE OF VASCULAR NICHE HOMEOSTASIS AND SUSCEPTIBILITY TO EMPHYSEMA. WE WILL USE CONDITIONAL MODELS TO MANIPULATE DKK1 OR IL13 EXPRESSION IN MICE ALLOWED TO AGE IN THE PRESENCE OR ABSENCE OF CIGARETTE SMOKE EXPOSURE BOTH IN VIVO AND IN VITRO. LASTLY, WE WILL TEST THAT LOSS OF ILC2 AND MVPC PROGENITORS PROMOTES LUNG AGING AND IS ASSOCIATED WITH COPD AND A TYPE-2 IMMUNE ENDOTYPE. THIS WORK WILL PROVIDE AN UNDERSTANDING OF PROGENITOR AGING, MECHANISMS BY WHICH LOSS OF MVPC AND ILC2 FUNCTION DRIVES VASCULAR REMODELING WITH A TH2 ENDOTYPE CONTRIBUTING TO LUNG AGING AND EMPHYSEMA.

MDS-ASSOCIATED SPLICEOSOME MUTATIONS REGULATE HOST DEFENSE - ABSTRACT MYELODYSPLASTIC SYNDROME (MDS) IS A HEMATOPOIETIC STEM CELL DISORDER CHARACTERIZED BY MYELOID CELL DIFFERENTIATION DEFECTS AND DYSPLASTIC BLOOD CELL PRODUCTION. THE MAJORITY OF MDS PATIENTS DIE OF DISEASE RELATED CAUSES, WITH INFECTION OR INFECTIOUS COMPLICATIONS BEING THE MOST COMMON CAUSE. WHILE IT IS KNOWN THAT MYELOID CELLS EXHIBIT FUNCTIONAL DEFECTS IN MDS PATIENTS, THE EXTENT AND CAUSE OF THESE DEFECTS REMAIN UNCLEAR AND THE CORRESPONDING EFFECTS ON HOST DEFENSE HAVE RECEIVED LIMITED STUDY. WITH THE ADVENT OF NEXT-GENERATION SEQUENCING TECHNOLOGY, ANALYSIS OF SOMATICALLY-ACQUIRED MUTATIONS IN PATIENT SAMPLES HAS BECOME A NORMAL PART OF CLINICAL PRACTICE IN MDS PATIENTS. INTERESTINGLY, THE MOST COMMON CLASS OF MUTATIONS FOUND IN MDS PATIENTS ARE MUTATIONS IN VARIOUS COMPONENTS OF THE SPLICEOSOME. WE HAVE DETERMINED THAT THESE MDS- ASSOCIATED SPLICEOSOME GENE MUTATIONS LEAD TO ALTERATIONS IN INNATE IMMUNE SIGNALING PATHWAYS AND COMPROMISE THE FUNCTION OF MYELOID CELLS IN MOUSE MODELS OF SPLICEOSOME-MUTATED MDS. THIS LEADS TO A SIGNIFICANT DEFECT IN HOST DEFENSE. BASED ON THESE PRELIMINARY STUDIES, WE HAVE HYPOTHESIZED THAT MDS PATIENTS WITH SPLICEOSOME MUTATIONS ARE AT AN INCREASED RISK OF INFECTION BECAUSE OF IMMUNE DYSFUNCTION IN THEIR MYELOID CELLS. TO TEST THIS HYPOTHESIS, WE WILL INVESTIGATE THE EFFECTS OF SPLICEOSOME MUTATIONS ON HOST DEFENSE USING: (1) EX VIVO STUDIES WITH MOUSE AND HUMAN NEUTROPHILS, (2) EX VIVO STUDIES WITH MOUSE AND HUMAN MACROPHAGES, AND (3) IN VIVO STUDIES IN MICE EXPRESSING MUTANT SPLICEOSOME GENES AND IN AN ANALYSIS OF CLINICAL DATA FROM PATIENTS WITH MDS. THESE STUDIES WILL DETERMINE THE MECHANISMS UNDERLYING HOST DEFENSE DEFECTS IN MDS PATIENTS WITH SPLICEOSOME MUTATIONS AND WILL PROVIDE IMPORTANT CLINICAL DATA ABOUT PATIENT RISK STRATIFICATION.

REDUCING FIBROBLAST PERSISTENCE IN PULMONARY FIBROSIS AS A MECHANISM OF RESOLUTION

ROLE OF BURN PIT-RELATED PARTICULATE MATTER IN DRIVING DEPLOYMENT-RELATED LUNG DISEASE

MACROPHAGE ENDOCYTOSIS IN RESOLVING LUNG INFLAMMATION

UNEARTHING INTERACTING NONTUBERCULOUS MYCOBACTERIAL, ENVIRONMENTAL, AND HOST DETERMINANTS OF LUNG DISEASE IN THE HAWAI'I ISLANDS

LUNG REPAIR BY TETRASPANIN INTERACTING PROTEIN IGSF3

MATERNAL PROGRAMMING OF THE STEM CELL-BASOPHIL AXIS FOR ASTHMA

BLACK LUNG

NOVEL INTEGRATIVE APPROACHES FOR DISEASE PHENOTYPING, UTILIZING RADIOMICS IN SARCOIDOSIS

DEVELOPMENT AND VALIDATION OF A PROGNOSTIC TRANSCRIPTOMIC SIGNATURE FOR CHRONIC HYPERSENSITIVITY PNEUMONITIS

FIBROBLAST RESISTANCE TO APOPTOSIS IN PULMONARY FIBROSIS

DNA INDUCTION OF NEUTROPHILIC ASTHMA - PROJECT SUMMARY NEUTROPHILIC ASTHMA IS A SUBTYPE OF SEVERE ASTHMA, WHICH HAS NO SAFE AND EFFECTIVE THERAPY. THERE IS AN UNMET NEED TO DELINEATE THE MECHANISM OF NEUTROPHILIC ASTHMA AND DEVELOP TARGETED AND EFFECTIVE THERAPIES. HOST CELL-DERIVED DNA IS PRESENT IN THE EXTRACELLULAR FLUID AND SERUM. DNA REPRESENTS A DANGER SIGNAL (DANGER- ASSOCIATE MOLECULAR PATTERN-DAMP) FOR HOST CELLS. RECENT STUDIES SUGGEST AN IMPORTANT ROLE FOR EXTRACELLULAR DNA IN MEDIATING VIRUS-INDUCED ASTHMA EXACERBATION. WE PRESENT ROBUST PRELIMINARY DATA DEMONSTRATING INCREASED LEVELS OF EXTRACELLULAR DNA, THE DNA SENSOR IFI16 (INTERFERON-GAMMA INDUCED PROTEIN-16) AND THE DNA-IFI16 PATHWAY-DRIVEN CYTOKINES/CHEMOKINES IN THE AIRWAYS FROM NEUTROPHILIC ASTHMA. WE DEVELOPED A MOUSE MODEL OF ASTHMA WHERE DNA INDUCES NEUTROPHILIC INFLAMMATION IN THE CONTEXT OF AN IL10-CONSTRAINED INFLAMED AIRWAY MILIEU. THIS PHENOTYPE REQUIRES THE PARTICIPATION OF THE IFI16 SIGNALING ADAPTER STING. BASED UPON THESE NOVEL PRELIMINARY RESULTS WE HYPOTHESIZE THAT EXTRACELLULAR DNA INDUCES NEUTROPHILIC ASTHMA THROUGH THE IFI16-STING PATHWAY IN THE PRESENCE OF SELECT IL10-SUPPRESSIVE TNFSFS. NEUTROPHIL EXTRACELLULAR TRAPS GENERATE DNA, WHICH ESTABLISHES A SELF-PERPETUATED MECHANISM OF NEUTROPHILIC ASTHMA. UNDER AIM 1 WE WILL EXAMINE THE RELEVANCE OF EXTRACELLULAR DNA FOR HUMAN NEUTROPHILIC ASTHMA. WE WILL STUDY THE GENERATION OF AIRWAY EXTRACELLULAR DNA, ACTIVATION OF IFI16 AND IL10-SUPPRESSIVE TNFSFS—TNF AND OX40L AND THEIR PATHOPHYSIOLOGICAL CONSEQUENCES IN THE AIRWAYS. WE WILL STUDY BRONCHOALVEOLAR LAVAGE (BAL), BRONCHIAL EPITHELIAL CELLS AND BIOPSY SPECIMENS FROM 3 STUDY GROUPS: 1) NEUTROPHILIC (WITH AND WITHOUT EOSINOPHILIC) ASTHMA; 2) NON-NEUTROPHILIC (WITH AND WITHOUT EOSINOPHILIC) ASTHMA; AND 3) DISEASE CONTROLS. WE WILL DELINEATE THE FUNCTION AND IMPORTANCE OF IFI16 FOR PRONEUTROPHILIC BIOMOLECULES IN A REDUCTIONIST MODEL IN DNA-TREATED AIRWAY MACROPHAGES AND BLOOD MONOCYTES. UNDER AIM 2 WE WILL STUDY THE MECHANISM OF EXTRACELLULAR DNA- INDUCED NEUTROPHILIC ASTHMA IN MICE. WE WILL ELUCIDATE THE ROLE OF IL10 AND TNFSFS (TNF AND OX40L) IN SWITCHING THE DNA-INDUCED DEFENSIVE PROGRAM TO A NEUTROPHILIC INFLAMMATION PROGRAM IN MOUSE AIRWAYS. WE WILL ESTABLISH THE ROLE OF IFI204 (THE MOUSE IFI16 ORTHOLOG) AND STING IN NEUTROPHILIC INFLAMMATION. WE WILL STUDY THE EFFECT OF REMOVAL OF EXTRACELLULAR DNA ON PERSISTENCE OF NEUTROPHILIC ASTHMA IN MICE. THIS PROJECT IS IMPORTANT BECAUSE IT UNCOVERS A NOVEL DNA-IFI16-STING MECHANISM OF NEUTROPHILIC ASTHMA AND ASSESSES THE THERAPEUTIC BENEFITS OF DNA AND STING INHIBITORS, AND DNA SCAVENGERS IN A PRECLINICAL STUDY.

GM-CSF, MACROPHAGES, AND SUSCEPTIBILITY TO MYCOBACTERIUM ABSCESSUS PULMONARY INFECTION - GM-CSF, MACROPHAGES, AND SUSCEPTIBILITY TO MYCOBACTERIUM ABSCESSUS PULMONARY INFECTION NONTUBERCULOUS MYCOBACTERIA (NTM) DO NOT CAUSE DISEASE IN HEALTHY INDIVIDUALS; HOWEVER, PEOPLE WITH CHRONIC AIRWAYS DISEASES ARE SUSCEPTIBLE TO DEVELOPING PULMONARY NTM INFECTIONS (PNTM), WHICH INCREASE SYMPTOM BURDEN AND ACCELERATE LUNG FUNCTION DECLINE. HOW THE HEALTHY LUNG CLEARS INHALED NTM AND WHY INDIVIDUALS WITH AIRWAYS DISEASE ARE AT INCREASED RISK OF DEVELOPING PNTM REMAIN POORLY UNDERSTOOD. OUR LONG TERM GOAL IS TO UNDERSTAND HOW MACROPHAGE HETEROGENEITY CONTRIBUTES TO CHRONIC LUNG DISEASES. THE OBJECTIVE OF THIS GRANT IS TO CHARACTERIZE EVENTS THAT OCCUR WHEN INHALED NTM INITIALLY INTERACT WITH RESPIRATORY TRACT MACROPHAGES, AND DETERMINE HOW THE CYTOKINE GRANULOCYTE MACROPHAGE COLONY STIMULATING FACTOR (GM-CSF) PROMOTES CLEARANCE OF NTM. DATA FROM HUMAN CASE REPORTS AND MICE LACKING GM-CSF (GM-CSFKO MICE) INDICATE THAT GM-CSF IS ESSENTIAL FOR IMMUNE CONTROL OF PNTM. WE HAVE CHOSEN TO STUDY THE ROLE OF GM-CSF IN SUSCEPTIBILITY TO MYCOBACTERIUM ABSCESSUS (MABSC) AS THIS SPECIES CAUSES A SIGNIFICANT PORTION OF PNTM, IS INCREASING IN PREVALENCE, AND IS ESPECIALLY CHALLENGING TO ERADICATE. OUR DATA DEMONSTRATE THAT GM-CSF CAN ACTIVATE MACROPHAGES TO KILL MABSC. IN LUNG AIRSPACES, MACROPHAGES CAN BE GROUPED INTO TWO MAIN CATEGORIES: RESIDENT ALVEOLAR MACROPHAGES (ALVMS) AND RECRUITED MONOCYTE-DERIVED MACROPHAGES (MDMS). ALVMS ARE CONSTITUTIVELY EXPOSED TO GM-CSF PRODUCED BY ALVEOLAR EPITHELIAL CELLS. IN CONTRAST, MOST MACROPHAGES IN AIRWAYS OF PEOPLE WITH CHRONIC AIRWAYS DISEASE ARE MDMS RECRUITED FROM THE BLOODSTREAM, A COMPARTMENT WITH LOW LEVELS OF GM- CSF. OUR CENTRAL HYPOTHESIS IS THAT MACROPHAGES MUST BE ACTIVATED BY GM-CSF TO ELIMINATE NTM, AND THAT MDMS RECRUITED TO THE AIRWAYS IN PEOPLE WITH CHRONIC AIRWAYS DISEASE ARE LESS EFFECTIVE AT KILLING NTM THAN ALVMS DUE TO INSUFFICIENT EXPOSURE TO GM-CSF. WE WILL INVESTIGATE THIS HYPOTHESIS IN THREE SPECIFIC AIMS. AIM 1 WILL DETERMINE THE MECHANISM BY WHICH GM-CSF ENHANCES MACROPHAGE KILLING OF MABSC, TESTING THE HYPOTHESIS THAT GM-CSF PROMOTES PHAGOSOMAL MATURATION AND PHAGOLYSOSOMAL ACIDIFICATION. USING CONDITIONAL KNOCKOUT MICE THAT LACK THE GM-CSF RECEPTOR ON DIFFERENT MACROPHAGE SUBSETS, AIM 2 WILL TEST THE HYPOTHESIS THAT RESIDENT ALVMS PROGRAMMED BY GM-CSF ARE THE ESSENTIAL CELLS RESPONSIBLE FOR CONTROL OF MABSC IN THE HEALTHY LUNG. AIM 3 WILL EMPLOY A MOUSE MODEL OF AIRWAY INFECTION USING MABSC-EMBEDDED AGAR BEADS TO TEST THE HYPOTHESIS THAT THERE IS INSUFFICIENT GM-CSF IN THE AIRWAYS TO ACTIVATE RECRUITED MDMS TO DEVELOP MYCOBACTERIOCIDAL PHENOTYPES. THESE STUDIES ARE INNOVATIVE, AS THEY WILL DETERMINE THE MECHANISMS BY WHICH GM-CSF ENHANCES MACROPHAGE KILLING OF MABSC, AND REVEAL HOW LOCATION IN THE LUNG INFLUENCES PHENOTYPES OF RECRUITED MDMS. THE RESULTS FROM THE PROPOSED RESEARCH WILL BE SIGNIFICANT, AS THEY WILL INFORM USE OF IMMUNE THERAPIES TO TREAT NON-RESOLVING INFECTION IN CHRONIC LUNG DISEASES, AND THEY COULD HELP PREDICT WHICH INDIVIDUALS WITH CHRONIC LUNG DISEASES ARE AT HIGHEST RISK FOR DEVELOPING PNTM.

PREDICTIVE MODELS OF BERYLLIUM SENSITIZATION AND CHRONIC BERYLLIUM DISEASE - PROJECT SUMMARY/ABSTRACT BERYLLIUM (BE) IS USED IN A SEVERAL INDUSTRIES AND US IS THE LARGEST EXPORTER OF BE IN THE WORLD. DESPITE MITIGATION STRATEGIES, BE EXPOSURE AT WORKPLACE RESULTS IN BE SENSITIZATION (BES) AND CHRONIC BERYLLIUM DISEASE (CBD). CBD IS AN IMPORTANT UNDERSTUDIED ORGAN-SPECIFIC IMMUNE-MEDIATED DISEASE CHARACTERIZED BY GRANULOMATOUS LUNG INFLAMMATION, FIBROSIS, AND DEATH. HENCE, CBD IS A PUBLIC HEALTH CONCERN RESULTING IN PROMULGATION OF NEW EXPOSURES STANDARDS RECENTLY. BES DEVELOPS IN UP TO 20% OF BE-EXPOSED INDIVIDUALS AND PROGRESSES TO CBD 50-100% OF THESE AT-RISK INDIVIDUALS. THE GOAL OF THIS STUDY IS TO DEFINE THE NOVEL LUNG COMPARTMENT-SPECIFIC GENE-PROTEIN PATHWAYS, WHICH WILL NARROW THE EXISTING GAP IN UNDERSTANDING OF BE LUNG DISEASE AND FORM THE BASIS OF A CLINICALLY VIABLE MODELS (CLASSIFIERS) TO IDENTIFY BES, CBD AND PREDICT PROGRESSION OF BES TO CBD. THIS PROJECTS HYPOTHESIS IS THAT SYSTEMATIC CHARACTERIZATION OF LUNG COMPARTMENT- SPECIFIC CHANGES WILL IDENTIFY NOVEL PATHWAYS LINKED TO BES, CBD AND THE PROGRESSION OF BES TO CBD. THE INVESTIGATORS POSIT THAT THE PROTEINS IN THESE PATHWAYS WOULD PROVIDE CLINICALLY VIABLE MODELS (CLASSIFIERS) THAT WILL DISCRIMINATE BETWEEN HEALTHY CONTROLS, BES, AND CBD. IN AIM 1, THE STUDY WILL DETERMINE THE SYSTEMS LEVEL, LUNG COMPARTMENT-SPECIFIC, GENE-PROTEIN CHANGES LINKED TO BES AND CBD BY ANALYZING BAL CELLS FROM A DISCOVERY COHORT (CBD=50, BES=50, HEALTHY CONTROLS=25). THIS AIM WILL IDENTIFY THE TRANSCRIPTIONAL AND GLOBAL PROTEIN CHANGES USING CONTEMPORARY HIGH-RESOLUTION MASS-SPECTROMETRY COUPLED WITH ADVANCED COMPUTATIONAL BIOLOGY AND BIOINFORMATICS. A COMBINATION OF SINGLE CELL RNA SEQUENCING AND INNOVATIVE EXPRESSION DECONVOLUTION WILL DEFINE CELL-SPECIFIC TRANSCRIPTIONAL CHANGES IMPLICATED IN BES AND CBD. FURTHERMORE, THE RESEARCH TEAM WILL INTEGRATE TRANSCRIPTION AND PROTEIN EXPRESSION TO IDENTIFY THE BIOLOGICAL PATHWAYS ASSOCIATED WITH BE-INDUCED LUNG DISEASE. THE STUDY USES THE SAME DISCOVERY COHORT, AND AN INDEPENDENT VALIDATION COHORT OF SUBJECTS ALREADY ENROLLED IN AIM 2, TO DEVELOP AND VALIDATE A COMPREHENSIVE BAL FLUID CLASSIFIER OF BES AND CBD. SPECIFICALLY, THE DISCOVERY COHORT WILL BE USED TO CONSTRUCT AND INTERNALLY VALIDATE A MULTICOMPONENT CLASSIFIER THAT DISCRIMINATES HEALTHY CONTROLS. IN ADDITION, THIS CLASSIFIER WILL BE EXTERNALLY VALIDATED IN THE VALIDATION COHORT OF CBD (N=50), BES (N=50), AND HEALTHY CONTROLS (N=25). IN AIM 3, THE STUDY EVALUATES THE STABILITY OF THE CLASSIFIER OF BES AND CBD AND DEVELOPS PREDICTIVE MODELS OF PROGRESSION FROM BES TO CBD IN SUBJECTS WITH LONGITUDINAL CLINICAL DATA AND BAL FLUID ALREADY AVAILABLE. A GROUP OF BES CASES WHO ON FOLLOW-UP DEVELOPED CBD WILL BE COMPARED TO BES CASES WHO DO NOT DEVELOP CBD TO BUILD A MODEL THAT PREDICTS BES PROGRESSION. AT THE COMPLETION OF THIS PROJECT, THE STUDY FINDINGS WILL BE POISED TO TRANSLATE TO CLINICAL CARE FOR THE RAPID DETECTION OF CBD AND BES. FURTHER, IT WILL IDENTIFY NOVEL PATHWAYS ASSOCIATED WITH BE LUNG DISEASE THAT CAN BE TESTED IN FUTURE STRUCTURE-FUNCTION STUDIES IN MODEL SYSTEMS OF BE LUNG DISEASE.

REGULATION OF PULMONARY INFLAMMATION

ALVEOLAR TYPE II CELL INNATE IMMUNE RESPONSE TO INFLUENZA

MOLECULAR CHARACTERIZATION OF PROGRESSIVE PULMONARY SARCOIDOSIS - PROJECT SUMMARY PULMONARY INVOLVEMENT OCCURS IN UP TO 90% OF SARCOIDOSIS CASES, THE DISEASE COURSE IS HETEROGENEOUS AND RESPIRATORY FAILURE IS THE LEADING CAUSE OF SARCOIDOSIS-RELATED MORTALITY IN THE US. MOST STUDIES UNDERTAKEN TO DATE IN SARCOIDOSIS COMPARE CASES TO HEALTHY OR DISEASE CONTROLS. THERE ARE KNOWLEDGE GAPS IN THE UNDERSTANDING OF SARCOIDOSIS SUBTYPES INCLUDING THE UNDERLYING MOLECULAR FEATURES, BIOLOGIC PATHWAYS AND MECHANISMS OF PROGRESSIVE PULMONARY SARCOIDOSIS DISEASE PROGRESSION. AS A RESULT, THERE ARE NO CLINICAL TOOLS AVAILABLE FOR EARLY PREDICTION OF PROGRESSIVE (P) SARCOIDOSIS TO ALLOW CLOSER CLINICAL FOLLOW-UP, EARLY TREATMENT AND TO PROVIDE FOCUS FOR RESEARCH. THE GOAL OF THIS PROJECT IS TO ENROLL SARCOIDOSIS PATIENTS TO DEFINE MOLECULAR CHARACTERISTICS OF P- SARCOIDOSIS THAT WILL (A) ENABLE EARLY IDENTIFICATION OF PATIENTS AT HIGHER RISK OF PROGRESSION. TO DEVELOP TOOLS FOR RAPID IDENTIFICATION AND PREDICTION OF P-SARCOIDOSIS PROGRESSION, THIS PROJECT WILL INVESTIGATE MOLECULAR SIGNATURES (PROTEINS AND GENES ) AND RELATED BIOLOGICAL PATHWAYS IDENTIFIED IN A LABORATORY MODEL OF SARCOIDOSIS GRANULOMAS AND IN PERIPHERAL BLOOD MONONUCLEAR CELLS (PBMCS) OF PATIENTS WITH P- VS NON-PROGRESSIVE (NP)- SARCOIDOSIS. (B) PROVIDE INSIGHTS INTO THE DIFFERENT BIOLOGICAL MECHANISMS DRIVING OUTCOMES IN PULMONARY SARCOIDOSIS. OUR PROMISING PILOT DATA IDENTIFIED PROTEIN AND TRANSCRIPTIONAL PATHWAYS ENGAGED DURING GRANULOMATOUS INFLAMMATION AND IN PMBCS IN PROXIES OF P- AND NP-SARCOIDOSIS. AIM 1 BUILDS ON PRELIMINARY FINDINGS, CHARACTERIZING DIFFERENCES IN P- AND NP-SARCOIDOSIS PROTEIN AND GENE PATHWAYS, LEVERAGING THE UNIQUE GRANULOMA MODEL. WE INCLUDE SINGLE CELL-RNA-SEQUENCING IN A SUBSET OF CASES WITH INNOVATIVE DIGITAL CYTOMETRY METHODS TO OBTAIN CELL- SPECIFIC PATHWAYS. AIM 2 WILL DEFINE PATHWAYS AND SIGNATURES FROM PBMCS, WHICH ARE RECRUITED AND CONTRIBUTE TO GRANULOMA DEVELOPMENT AND PERSISTENCE AS WELL AS FEATURES THAT ARE SHARED AND DISTINCT FROM THE MODEL. AIM 3 WILL LEVERAGE FINDINGS FROM PMBCS AND THE IN VITRO MODEL AND CLINICAL DISEASE MANIFESTATIONS USING INNOVATIVE BIOINFORMATICS APPROACHES TO CONSTRUCT AND INTERNALLY VALIDATE A COMPREHENSIVE CLASSIFIER THAT INCORPORATES PROTEINS, CLINICAL VARIABLES AND PATIENT-REPORTED OUTCOMES IN A DISCOVERY COHORT. THE INVESTIGATORS WILL VALIDATE THIS CLASSIFIER IN AN INDEPENDENT VALIDATION COHORT OF SUBJECTS ALREADY ENROLLED BY THE RESEARCH TEAM IN PRIOR NIH FUNDED STUDIES. THE STUDY USES NOVEL APPROACHES TO CHARACTERIZE P-SARCOIDOSIS AND A DIVERSE RESEARCH TEAM LED BY MPIS WITH COMPLEMENTARY EXPERTISE. THE INTEGRATION OF SARCOIDOSIS INVESTIGATORS FROM DIFFERENT LOCATIONS ACROSS THE COUNTRY WILL ADVANCE AND SUSTAIN THE SARCOIDOSIS RESEARCH THROUGH HIGHLY INNOVATIVE TRANSLATIONAL STUDIES IMPLEMENTING STATE-OF-THE-ART CLINICAL AND BIOLOGICAL SYSTEMS-LEVEL STUDIES IN A HIGH-PRIORITY AREA OF RESEARCH IN SARCOIDOSIS. THESE STUDIES WILL PROVIDE THE FOUNDATION FOR FUTURE STUDIES AIMED AT EVALUATING THIS CLASSIFIER IN LONGITUDINAL AND PATIENT CARE STUDIES TO IDENTIFY AN AT-RISK SARCOIDOSIS POPULATION THAT MAY REQUIRE CLOSER FOLLOW UP AND EARLIER TREATMENT, AND RESEARCH PERTAINING TO THE IDENTIFICATION OF NOVEL MECHANISTIC PATHWAYS AND THERAPEUTIC TARGETS CONTRIBUTING TO P-SARCOIDOSIS.

USE OF BLINDED TAPERING FOR HYPNOTIC DISCONTINUATION

MOTIVATING ADHERENCE TO CPAP IN OBSTRUCTIVE SLEEP APNEA

CONTRIBUTION OF PULMONARY IONOCYTES AND NEUROENDOCRINE CELLS TO ION TRANSPORT-MEDIATED AIRWAY SURFACE LIQUID MAINTENANCE - PROJECT SUMMARY/ABSTRACT REGULATION OF ION TRANSPORT ACROSS THE EPITHELIUM IS VITAL TO THE HEALTH AND FUNCTION OF THE AIRWAYS. ACTIVE AND PASSIVE ION AND WATER MOVEMENT IS RESPONSIBLE FOR MAINTAINING A PERICILIARY FLUID LAYER THROUGH WHICH CILIA CAN MOVE MUCUS AND PROTECT THE AIRWAYS AGAINST PARTICLES, IRRITANTS, AND PATHOGENS. WHILE THERE IS CLEAR EVIDENCE OF DISEASE CAUSED BY SPECIFIC ION CHANNEL MUTATIONS (CFTR) AS WELL ACQUIRED ION CHANNEL DYSFUNCTION DURING/CONTRIBUTING TO DISEASE SEVERITY (COPD, ASTHMA), THE CONTRIBUTION OF INDIVIDUAL CELL TYPES OF THE PULMONARY EPITHELIUM TO ION TRANSPORT MAINTENANCE IS POORLY UNDERSTOOD. OUR EXPERIMENTAL EVIDENCE DEMONSTRATES THAT ALTERATIONS IN THE FREQUENCIES OF PULMONARY IONOCYTES AND NEUROENDOCRINE CELLS IN AIRWAY EPITHELIAL CULTURES RESULT IN DRAMATIC DIFFERENCES IN ION TRANSPORT PROPERTIES. ALTOGETHER, THESE CELL TYPES, ALONG WITH TUFT-LIKE CELLS, COMPRISE ABOUT 0.8% OF THE TOTAL CELL NUMBERS IN THE HUMAN TRACHEAL EPITHELIUM. PULMONARY IONOCYTES ARE A NEWLY IDENTIFIED CELL TYPE IN THE AIRWAYS WITH EXTREMELY HIGH EXPRESSION OF MANY ION CHANNELS WITH VITAL FUNCTIONS IN THE RESPIRATORY TRACT. THE DRAMATIC IMPACT OF IONOCYTES AND PULMONARY NEUROENDOCRINE CELLS ON THE ION TRANSPORT PROPERTIES OF HUMAN AIRWAY CULTURES DESPITE THEIR RARE FREQUENCIES LED TO OUR HYPOTHESIS THAT THESE CELL TYPES ARE HIGHLY INVOLVED IN THE REGULATION AND MAINTENANCE OF ION TRANSPORT BALANCE ACROSS THE ENTIRE EPITHELIUM AND ARE ABLE TO IMPACT ION TRANSPORT FUNCTIONS IN OTHER CELL TYPES THROUGH INTERCELLULAR COMMUNICATION. BY UTILIZING NOVEL MURINE MODELS AND APPLYING INNOVATIVE TECHNIQUES TO PRIMARY MURINE AND HUMAN AIRWAY EPITHELIAL CELLS, THE RESEARCH OUTLINED IN THIS PROPOSAL WILL DEFINE THE CONTRIBUTION OF THESE RARE CELL TYPES TO THE OVERALL ION TRANSPORT FUNCTIONS OF THE AIRWAY EPITHELIUM AND WILL ALSO DEFINE THE ROLE OF CFTR IN THESE CELLS. WE WILL THEREBY FULLY CHARACTERIZE THE SIGNIFICANCE OF PULMONARY IONOCYTES AND NEUROENDOCRINE CELLS TO ION TRANSPORT-MEDIATED AIRWAY SURFACE LIQUID REGULATION AND IDENTIFY POTENTIAL CELL AND ION CHANNEL TARGETS FOR THERAPEUTIC TREATMENT OF COMMON AIRWAY DISEASES.

NATURALLY OCCURRING T REGULATORY CELLS CONTROL AIRWAY HYPERRESPONSIVENESS

ATOPIC DERMATITIS RESEARCH NETWORK (ADRN) CLINICAL RESEARCH CENTER

STEROID RESISTANCE OF AIRWAY ILC2S

TEMPORAL REGULATION OF PULMONARY INFLAMMATION BY MYD88 ALTERNATIVE PRE-MRNA SPLICING

EXPOSURE IN EPIGENETIC REGULATION OF IMMUNE RESPONSE IN CBD

PRECISION INTERVENTIONS FOR SEVERE AND/OR EXACERBATION PRONE ASTHMA (PRECISE)

ESTABLISHMENT OF REPOSITORY FOR CHRONIC BERYLLIUM DISEASE

LUNG MSC REGULATE ANGIOGENESIS AND REPAIR DURING FIBROSIS

LUNG REGENERATION AND THE STEM CELL NICHE

EPIGENETIC REGULATION OF ALTERED T-CELL IMMUNITY IN SARCOIDOSIS

THE EFFECT OF MYCOPLASMA ON CHRONIC ASTHMA

TRANSCRIPTIONAL CONTROL OF T CELL FUNCTION - PROJECT SUMMARY T CELLS RESPONDING TO CANCER DEVELOP A HYPORESPONSIVE PHENOTYPE, CHARACTERIZED BY HIGH LEVELS OF INHIBITORY RECEPTOR EXPRESSION, LOW CYTOKINE PRODUCTION, AND A FAILURE TO CONTROL TUMOR GROWTH. WE HAVE PREVIOUSLY IDENTIFIED A TRANSCRIPTIONAL PROGRAM INDUCED BY PERSISTENT ANTIGEN STIMULATION THAT WAS SHARED BY MOUSE AND HUMAN T CELLS RESPONDING TO CHRONIC VIRAL INFECTIONS AND TUMORS. WE LINKED THIS PROGRAM TO ACTIVATION OF THE NUCLEAR RECEPTOR SUBFAMILY 4 GROUP A (NR4A) TRANSCRIPTION FACTORS (TFS), WHICH ARE POTENTLY INDUCED DURING CHRONIC STIMULATION. WE ALSO IDENTIFIED NR4A-SENSITIVE REGULATORY ELEMENTS THAT BECAME ACCESSIBLE IN T CELLS RESPONDING TO CHRONIC VIRAL INFECTION AND TUMORS. WE FOUND THAT NR4A TFS CONCOMITANTLY PROMOTED INHIBITORY RECEPTOR EXPRESSION AND LIMITED CYTOKINE PRODUCTION, LEADING TO AN EXHAUSTED T CELL STATE THAT REDUCED CHIMERIC ANTIGEN RECEPTOR EXPRESSING T (CAR-T) CELL ACTIVITY AGAINST SOLID TUMORS. CAR-T CELLS ENGINEERED TO LACK ALL THREE NR4A TFS DIFFERENTIATED TO A UNIQUE POPULATION WITH POTENT ANTI-TUMOR ACTIVITY COMPARED TO WILD-TYPE T CELLS. WE CONNECTED THIS IMPROVED FUNCTION TO INCREASED EXPRESSION AND ACTIVITY OF BASIC LEUCINE ZIPPER (BZIP) TFS IN NR4A-DEFICIENT T CELLS THAN IN WILD-TYPE. IN NEW PRELIMINARY STUDIES, WE FOUND THAT CAR-T CELLS WITH A PARTIAL LOSS OF NR4A TFS ALSO PROVIDED BETTER PROTECTION THAN WILD-TYPE CAR-T CELLS, BUT HAD A UNIQUE “EXHAUSTION RESISTANT” PHENOTYPE THAT ARE POISED FOR EFFECTOR FUNCTION AND HAVE INCREASED POTENTIAL FOR LONG TERM SURVIVAL. IN THIS APPLICATION, WE WILL TEST THE HYPOTHESIS THAT NR4A AND BZIP TFS CAN BE “TUNED” TO CONTROL THE TRANSCRIPTIONAL PROGRAMS AND FUNCTION OF “EXHAUSTION RESISTANT” CAR-T CELLS IN TUMORS. IN AIM 1, WE WILL DETERMINE THE IMPACT OF THERAPEUTIC INTERVENTIONS, USING ANTIBODIES OR SMALL MOLECULES, ON THE FUNCTION AND SURVIVAL OF “EXHAUSTION RESISTANT” CAR-T CELLS. IN AIM 2, WE WILL DETERMINE THE CONTRIBUTION OF NR4A REGULATED BZIP TFS TO THE FUNCTION OF NR4A-DEFICIENT CAR-T CELLS AND THEIR EFFECTS ON TRANSCRIPTIONAL PROGRAMS IN “EXHAUSTION RESISTANT” CAR-T CELLS. IN AIM 3, WE WILL DEFINE THE EFFECTS OF NR4A TFS ON BZIP TF ACTIVITY AT REGULATORY ELEMENTS AND IDENTIFY CHROMATIN ASSOCIATED CO- FACTORS THAT MAY BE TARGETED TO CONTROL THE FUNCTION OF “EXHAUSTION RESISTANT” CAR-T CELLS. THE EXPECTED OUTCOME OF OUR PROPOSED STUDIES IS A COMPREHENSIVE UNDERSTANDING OF THE EFFECTS OF NR4A AND BZIP TFS ON CAR-T CELLS IN TUMORS, THROUGH WHICH WE WILL IDENTIFY PRACTICAL STRATEGIES TO “TUNE” THE FUNCTION OF CAR-T CELLS FOR THERAPEUTIC BENEFIT BY CONTROLLING TRANSCRIPTIONAL PROGRAMS.

ACTIVATION OF MATRIX METALLOPROTEINASE-9 IS ESSENTIAL TO OVERCOME FAILED FIBROSIS RESOLUTION - PROJECT SUMMARY A KEY FEATURE OF INTERSTITIAL LUNG DISEASES (ILDS), INCLUDING IDIOPATHIC PULMONARY FIBROSIS (IPF), IS THE EXCESSIVE DEPOSITION OF EXTRACELLULAR MATRIX (ECM) AND SCAR TISSUE. FIBROBLASTS PERSIST IN FIBROTIC LUNGS AND LAY DOWN MATRIX, CONTRIBUTING TO A PROGRESSIVE AND PERSISTENT PHENOTYPE IN PATIENTS. MATRIX METALLOPROTEINASES (MMPS) ARE ENZYMES THAT CLEAVE AND BREAK DOWN ECM DURING WOUND REPAIR AND THE DEGRADATION AND REMOVAL OF SCAR TISSUE IS ESSENTIAL FOR FIBROSIS RESOLUTION. THE EXPRESSION OF MMP-9 IS INCREASED IN THE LUNGS OF IPF PATIENTS. HOWEVER, MMP-9 IS NOT PRESENT IN THE ACTIVE FORM RENDERING IT UNABLE TO CLEAVE COLLAGENS. OUR PRELIMINARY DATA IN MMP-9 DEFICIENT MICE SUPPORT THAT ACTIVE MMP-9 IS NECESSARY FOR FIBROSIS RESOLUTION. WE THEREFORE ASKED IF MMP-9 ACTIVATION IS INHIBITED IN IPF LUNGS LIMITING ITS ABILITY TO SUCCESSFULLY INITIATE REPAIR THROUGH MATRIX DEGRADATION. MMP-9 IS ACTIVATED FROM ITS LATENT-FORM THROUGH A SERIES OF ACTIVATING ENZYMES BEGINNING WITH UROKINASE PLASMINOGEN ACTIVATING ENZYME (UPA). UPA CLEAVES PLASMINOGEN INTO PLASMIN WHICH IN TURN IS A MAJOR ACTIVATOR OF MMP-9. HOWEVER, IN IPF, PLASMINOGEN ACTIVATOR INHIBITOR-1 (PAI-1) INHIBITS UPA THUS INHIBITING THE ACTIVATION CASCADE NEEDED FOR DOWNSTREAM MMP-9-ACTIVATION AND MATRIX DEGRADATION. DUE TO THE POOR QUALITY AND SIGNIFICANTLY REDUCED LIFE EXPECTANCY ASSOCIATED WITH ILDS, IT IS BECOMING INCREASINGLY IMPORTANT TO IDENTIFY MOLECULAR PATHWAYS THAT ARE TARGETABLE FOR THERAPEUTIC INTERVENTION. THIS PROPOSAL SEEKS TO ADDRESS THIS UNMET NEED BY INVESTIGATING THE CENTRAL HYPOTHESIS THAT MMP-9 ACTIVATION BY PLASMIN IS NECESSARY FOR FIBROSIS RESOLUTION AND THAT THIS PATHWAY CAN BE INDUCED THROUGH BENEFICIAL TNF-A SIGNALING. BASED ON OUR ROBUST PRELIMINARY STUDIES, WE PROPOSE THREE SPECIFIC AIMS TO TEST THIS CENTRAL HYPOTHESIS. SPECIFIC AIM 1 WILL TEST THE HYPOTHESIS THAT BENEFICIAL TNF-A SIGNALING INCREASES UROKINASE AND EXPRESSION OF MMP-9 IN FIBROBLASTS. THIS WILL BE TESTED USING GENETIC APPROACHES TO DETERMINE IF CONDITIONAL DELETION OF MMP-9 IN FIBROBLASTS IS SUFFICIENT TO PREVENT FIBROSIS RESOLUTION IN A SPONTANEOUSLY RESOLVING FIBROSIS MODEL AND IF EXOGENOUS TNF-A IS SUFFICIENT TO ACTIVATE THE UPA/PLASMINOGEN/MMP-9 PATHWAY, INDUCING RESOLUTION IN A NON-RESOLVING FIBROSIS MODEL. SPECIFIC AIM 2 WILL TEST THE HYPOTHESIS THAT MMP-9 ACTIVATION BY PLASMIN CONTRIBUTES TO THE RESOLUTION OF BLEOMYCIN-INDUCED FIBROSIS. THIS WILL BE TESTED THROUGH THE GENERATION OF A NON-CLEAVABLE MMP-9 MUTANT AND THROUGH PHARMACOLOGICAL INHIBITION OF MMP-9 IN VIVO. SPECIFIC AIM 3 WILL TEST THE HYPOTHESIS THAT A CLINICAL FORMULATION OF RECOMBINANT UROKINASE (KINLYTICTM) PROMOTES THE RESOLUTION OF ESTABLISHED PULMONARY FIBROSIS BY ACTIVATING PRO-MMP-9 THROUGH THE PLASMINOGEN/PLASMIN CASCADE. THIS WILL BE TESTED BY TREATING MICE WITH PERSISTENT FIBROSIS WITH RECOMBINANT UROKINASE TO INDUCE PLASMIN AND MMP-9 ACTIVATION AS A MECHANISM OF FIBROSIS RESOLUTION. THE PROPOSED STUDIES WILL PROVIDE A NOVEL UNDERSTANDING ABOUT HOW ACTIVATING AN ANTI-FIBROTIC PATHWAY (UPA/PLASMINOGEN/MMP-9) MAY CONTRIBUTE TO THE RESOLUTION OF FIBROSIS. FURTHERMORE, THE OUTCOMES OF THESE PRE-CLINICAL, THERAPEUTIC STUDIES WILL SIGNIFICANTLY IMPACT OUR UNDERSTANDING OF THE MECHANISMS THAT CONTROL FIBROSIS RESOLUTION.

IRAK-M IN LUNG DEFENSE AGAINST RHINOVIRUS INFECTION

CHECKPOINT FUNCTION OF PTPALPHA IN PATHOLOGICAL FIBROGENESIS IN THE LUNG

MECHANISMS OF STEROID RESISTANT ASTHMA

REVERSAL OF INFLAMMATORY PROCESSES IN CGD

INNATE EFFECTORS AND TYPE-2 INFLAMMATION

ROLE OF GPR116 IN ALVEOLAR HOMEOSTASIS

REGULATION OF B CELL DEVELOPMENT AND FUNCTION BY ZFP521

SOMATIC HYPERMUTATION IN SLE

ROLE OF MEK1 IN T CELL FUNCTION IN ASTHMA

ADAM17-NRG-1 SIGNALING IN ACUTE LUNG INJURY

REGULATION OF INFLAMMATION BY PULMONARY COLLECTINS

PHOSPHOLIPID DYNAMICS IN MEMBRANE ASSEMBLY

FUNCTIONAL ANERGY IN B CELLS

IMMUNOSUPPRESSIVE INJURIOUS EFFECTS OF E-CIGARETTES ON HUMAN LUNG PARENCHYMA

GENE RESPONSES IN BASOPHIL-MEDIATED ALLERGIC RESPONSES

REGULATION OF B CELL IDENTITY AND LINEAGE PROGRESSION

THIS RESEARCH WILL TEST THE HYPOTHESIS THAT HIGHER LEVELS OF ENDOTOXIN EXPOSURE CAUSE PERSISTENT PROBLEMATIC ASTHMA AND THAT KEY ENVIRONMENTAL (OZON

INTERACTION OF FIBROBLASTS WITH CELL CORPSES INCREASES COLLAGEN SYNTHESIS DURING LUNG REPAIR - PROJECT SUMMARY REPAIR AFTER INJURY IS A FUNDAMENTAL BIOLOGIC PROCESS THAT IS CRITICAL FOR MAINTAINING LUNG HEALTH. FIBROBLASTS MUST PERFORM KEY PRO-REPAIR FUNCTIONS AFTER INJURY INCLUDING PROLIFERATION AND SYNTHESIS OF NEW EXTRACELLULAR MATRIX, PARTICULARLY COLLAGENS, THAT SCAFFOLD WOUND CLOSURE. DEAD CELLS (COLLECTIVELY TERMED CELL CORPSES) ARE PRODUCED DURING LUNG INJURY AND INFLAMMATION AND HAVE BEEN SHOWN TO PROMOTE THE TRANSITION FROM INFLAMMATION TO REPAIR. PRIOR RESEARCH HAS FOCUSED ON UNDERSTANDING MACROPHAGE INTERACTIONS WITH APOPTOTIC CORPSES. HOWEVER, NON- PROFESSIONAL PHAGOCYTES INCLUDING FIBROBLASTS CAN ALSO INTERACT WITH CELL CORPSES AND THE CONSEQUENCE OF THESE INTERACTIONS IN THE LUNG HAS NOT BEEN STUDIED. THIS PROPOSAL SEEKS TO ADDRESS UNKNOWNS REGARDING FIBROBLAST- CORPSE INTERACTIONS AND THEIR ROLE IN HEALTHY LUNG REPAIR. WE HAVE FOUND THAT RECOGNITION OF CELL CORPSES BY FIBROBLASTS CAUSES FIBROBLASTS TO INCREASE COLLAGEN PROTEIN SYNTHESIS. OUR PRELIMINARY DATA SUPPORT A MECHANISM WHERE THE PHOSPHATIDYLSERINE RECEPTOR AXL BINDS CELLS CORPSES, ACTIVATES POLYAMINE METABOLITE SYNTHESIS, INCREASING INTRACELLULAR SPERMIDINE LEVELS, WHICH FUEL HYPUSINATION. HYPUSINATION OCCURS ON A SINGLE PROTEIN: THE RIBOSOMAL SUBUNIT EIF5A. WHEN HYPUSINATED, EIF5A IS ABLE TO STABILIZE TRANSLATION OF POLY-PROLINE-REPEAT PROTEINS INCLUDING COLLAGEN. HYPUSINATION IS ALSO IMPORTANT FOR CELL PROLIFERATION. BLOCKING POLYAMINE SYNTHESIS PREVENTED HYPUSINATION OF EIF5A IN RESPONSE TO CORPSE RECOGNITION. BLOCKING AXL OR HYPUSINE-EIF5A PREVENTED THE INCREASE IN COLLAGEN IN RESPONSE TO CORPSE RECOGNITION. WE ALSO SHOW THAT, IN VIVO, HYPUSINE-EIF5A IS INCREASED IN FIBROBLASTS AFTER LUNG INJURY AND THAT COLLAGEN DEPOSITION AND ALVEOLAR-CAPILLARY REPAIR ARE REDUCED WHEN HYPUSINATION IS BLOCKED WITH THE INHIBITOR GC7 OR THROUGH TARGETED DELETION OF THE HYPUSINATION ENZYME DEOXYHYPUSINE SYNTHASE IN FIBROBLASTS. THIS LED US TO HYPOTHESIZE THAT INTERACTION OF FIBROBLASTS WITH CELL CORPSES IS A CENTRAL CUE THAT ACTIVATES FIBROBLAST PROLIFERATION AND COLLAGEN SYNTHESIS FOLLOWING LUNG INJURY, FACILITATING REPAIR. WE WILL USE THREE AIMS TO TEST IF 1) CORPSE RECOGNITION VIA AXL TRIGGERS A TRANSLATIONALLY-REGULATED INCREASE IN COLLAGEN PROTEIN, 2) CORPSE RECOGNITION DRIVES POLYAMINE SYNTHESIS THAT INCREASES SPERMIDINE THAT INCREASES HYPUSINE-EIF5A AND STABILIZES COLLAGEN TRANSLATION, AND 3) TO FULLY ACTIVATE PRO-REPAIR PROGRAMMING FIBROBLASTS REQUIRE THE ABILITY TO SENSE CORPSES VIA AXL, SYNTHESIZE POLYAMINES VIA ORNITHINE DECARBOXYLASE, AND HYPUSINATE EIF5A VIA DEOXYHYPUSINE SYNTHASE IN ORDER TO PROLIFERATE AND SYNTHESIZE NEW EXTRACELLULAR MATRIX AFTER LUNG INJURY.

INNATE IMMUNITY AND DEPLOYMENT-RELATE LUNG DISEASES

MECHANISMS OF MYOFIBROBLAST DEATH IN PULMONARY FIBROSIS

IMPROVED PREDICTION OF RESPONSE TO ASTHMA MEDICATION USING SMALL MOLECULES

SPLUNC1 PROTEIN IN HOST DEFENSE AGAINST MYCOPLASMA PNEUMONIAE INFECTION

CERAMIDE MEDIATES COVID-19 VASCULAR INJURY AND ARDS

THE IMMUNOREGULATORY ROLE OF ALVEOLAR MACROPHAGES IN CHRONIC BERYLLIUM DISEASE

AIRWAY TH2TH17 CELLS IN REFRACTORY ASTHMA

TOXIN-INDUCED ALTERNATIVE MRNA SPLICING IN TOLL-LIKE RECEPTOR SIGNALING PATHWAYS

THE ROLE OF BATF IN ALLERGIC INFLAMMATION AND ANTI-HELMINTH IMMUNITY

FOCAL ADHESION MODULATION OF IL-1 SIGNALING: IMPORTANCE IN PULMONARY FIBROSIS

HISTONE ARGININE DEMETHYLATION THROUGH CLEAVAGE

TARGETING OXIDATIVE STRESS IN CHRONIC BERYLLIUM DISEASE

NATIONAL JEWISH HEALTH COFAR CLINICAL RESEARCH UNIT

PROSPECTIVE LONGITUDINAL ASSESSMENT OF CULTURE-INDEPENDENT MOLECULAR AIRWAY MARKERS OF NONTUBERCULOUS MYCOBACTERIA

SELENOCYANATE AS A NOVEL TREATMENT OF CYSTIC FIBROSIS LUNG DISEASE

ENHANCING HYPNOTIC MEDICATION DISCONTINUATION IN PRIMARY CARE THROUGH SUPERVISED MEDICATION TAPERING AND DIGITAL COGNITIVE BEHAVIORAL INSOMNIA THERAPY - PROJECT SUMMARY/ABSTRACT TREATMENT-SEEKING INSOMNIA SUFFERERS MOST OFTEN PRESENT IN PRIMARY CARE WHERE THEIR FIRST AND USUALLY ONLY TREATMENT IS A PRESCRIPTION HYPNOTIC MEDICATION. MORE THAN 65% OF INDIVIDUALS PRESCRIBED HYPNOTICS USE THEM FOR MORE THAN A YEAR, AND MORE THAN 30% REMAIN ON THEM FOR MORE THAN FIVE YEARS. SUCH AGENTS MAY BE USEFUL FOR ACUTE INSOMNIA AND CERTAIN CASES WITH CHRONIC SLEEP DIFFICULTIES, BUT PROLONGED HYPNOTIC USE CAN LEAD TO DEPENDENCY AND INCREASED MORBIDITY (E.G., FALLS, COGNITIVE/DRIVING IMPAIRMENTS). REDUCING OR DISCONTINUING HYPNOTICS AFTER PROLONGED USE IS A CHALLENGING TASK FOR BOTH PRESCRIBING PHYSICIANS AND THE PATIENTS WHO USE THEM. ALTHOUGH EVIDENCED-BASED PHYSICIAN-SUPERVISED MEDICATION TAPERING (SMT) PROTOCOLS HAVE SHOWN EFFICACY, SUCH INTERVENTIONS HAVE YET TO BE DISSEMINATED WIDELY IN PRIMARY CARE. MOST PRIMARY CARE PROVIDERS (PCPS) ARE WILLING TO REFER THEIR INSOMNIA PATIENTS TO ALTERNATIVE EVIDENCE-BASED NON-DRUG TREATMENTS SUCH AS COGNITIVE BEHAVIORAL INSOMNIA THERAPY (CBTI), BUT SUCH TREATMENT IS OFTEN DIFFICULT TO ACCESS OUTSIDE OF SPECIALTY SLEEP CENTERS. GIVEN THIS GAP BETWEEN RESEARCH AND CLINICAL PRACTICE, THERE IS A PRESSING NEED TO DEVELOP AND VALIDATE COST-EFFECTIVE INTERVENTIONS TO FACILITATE THE MANAGEMENT OF INSOMNIA AND HYPNOTIC TAPERING IN PRIMARY CARE. IN RESPONSE TO PAR-20-183, THIS APPLICATION HAS BEEN CAREFULLY DESIGNED TO ADDRESS THESE ISSUES. WE WILL CONDUCT A LARGE RANDOMIZED TRIAL TO COMPARE THE COMBINED DIGITAL CBT (DCBTI)/SMT INTERVENTION, TO THE SMT INTERVENTION DELIVERED ALONE FOR PRODUCING HYPNOTIC DISCONTINUATION AND INSOMNIA SYMPTOM IMPROVEMENT. A SAMPLE OF 430 HYPNOTIC-RELIANT PATIENTS DRAWN FROM 8-10 PRIMARY CARE CLINICS WITHIN A PRACTICE-BASED RESEARCH NETWORK AFFILIATED WITH THE UNIVERSITY OF COLORADO MEDICAL SCHOOL IN AURORA, COLORADO WILL SERVE AS STUDY PARTICIPANTS. THE MAIN OBJECTIVE OF THE PROJECT IS TO COMPARE THE PERFORMANCE OF DCBTI+SMT WITH SMT USED ALONE FOR ACHIEVING HYPNOTIC REDUCTION/DISCONTINUATION AND INSOMNIA SYMPTOM IMPROVEMENT. IN ADDITION, WE WILL INCORPORATE AN EFFECTIVENESS-IMPLEMENTATION ASSESSMENT INTO THE RCT TO IDENTIFY PATIENT- PROVIDER- AND SYSTEM-LEVEL FACTORS THAT MAY IMPACT ADOPTION, IMPLEMENTATION AND MAINTENANCE OF THE TYPES OF INTERVENTIONS TESTED. FINDINGS FROM THE EFFECTIVENESS/IMPLEMENTATION SHOULD HELP DESIGN FUTURE TRIALS OF IMPLEMENTATION STRATEGIES IDENTIFIED HEREIN TO PROMOTE DISSEMINATION OF DCBTI AND SMT INTERVENTIONS INTO PRIMARY CARE SHOULD THESE TREATMENTS PROVE EFFECTIVE IN THE CURRENT TRIAL. WE ALSO WILL GATHER EXPLORATORY DATA TO DETERMINE WHO RESPONDS BEST TO DCBTI/SMT. THIS STUDY WILL PROVIDE NEW AND USEFUL INFORMATION ABOUT THE FEASIBILITY, CLINICAL UTILITY, AND PATIENT-, PROVIDER-, AND SYSTEM-LEVEL ACCEPTABILITY OF THESE INTERVENTIONS TO MANAGE INSOMNIA AND REDUCE/ELIMINATE HYPNOTIC USE IN PRIMARY CARE. THIS PROJECT SHOULD SERVE AS A NECESSARY FIRST STEP TOWARD THE EVENTUAL DISSEMINATION OF ACCESSIBLE, COST-EFFECTIVE STRATEGIES TO MANAGE A SIGNIFICANT PUBLIC HEALTH PROBLEM AND IMPROVE THE QUALITY OF LIFE OF MILLIONS OF CHRONIC USERS OF CONTROLLED-SUBSTANCE SLEEP AIDS.

NOVEL FUNCTION OF MUC18: AMPLIFICATION OF INFLAMMATION IN ALLERGIC LUNGS

SPROUTY-2 REGULATION OF SIGNALING IN ASTHMA

EPIGENETIC REGULATION OF IMMUNITY IN ALPHA-1 ANTI-TRYPSIN DEFICIENCY

METABOLOME AND PROTEOME PROFILES OF EMPHYSEMA AND AIRWAY DISEASE

ADAPTIVE GLUTATHIONE RESPONSES TO CIGARETTE SMOKE IN COPD

EFFECTS OF ALLERGIC INFLAMMATION ON TLR2 SIGNALING AND MYCOPLASMA INFECTION

DUSP1 AS A THERAPEUTIC TARGET IN FIBROPROLIFERATIVE ACUTE LUNG INJURY

GAMMA/DELTA T CELLS IN AUTOIMMUNE KERATITIS

FUNCTION AND REGULATION OF EARLY B CELL FACTOR (EBF)

DIRECT AND BONE-MARROW MEDIATED EFFECTS OF ADIPOSE STEM CELLS IN EMPHYSEMA

REGULATION AND FUNCTION OF CYTOSOLIC PHOSPHOLIPASES A2 IN LUNG CELLS

DEFICIENT SLEEP LUNG FUNCTION, AND FUNCTIONAL OUTCOMES IN ADOLESCENTS WITH ASTHMA

ANIONIC SURFACTANT LIPID REGULATION OF INFLAMMATION AND INFECTION IN THE LUNG

FOSTERING COLLABORATIONS AND THE CAREER OF A NEWLY RECRUITED PULMONARY SCIENTIST

VIRAL-INDUCED ADAPTATION OF NEUTROPHIL RESPONSE IN ARDS

SPHINGOLIPIDS AND INFLAMMATORY BIOMARKERS OF COPD PROGRESSION IN HEALTHY SMOKERS

1/2-SEQUENCED THERAPIES FOR COMORBID AND PRIMARY INSOMNIAS

INFLUENCE OF MATERNAL IMMUNITY ON THE RESPONSE OF THE NEWBORN TO RSV INFECTION

WNT REGULATION OF TRACHEOBRONCHIAL REGENERATION

PHOSPHOLIPID-PROTEIN INTERACTIONS REGULATING VIRAL INFECTION

RADIATION EXPOSURE SCREENING AND EDUCATION PROGRAM

NOVEL ROLES FOR LYSOPHOSPHOLIPPIDS IN EUKARYOTIC MEMBRANE BIOGENESIS AND TURNOVER

MAST CELL LINEAGE COMMITMENT AND FUNCTION - PROJECT SUMMARY THE INCIDENCE OF MAST CELL (MC)-MEDIATED DISEASES, INCLUDING ALLERGIC DISEASES AND MAST CELL ACTIVATION SYNDROME (MCAS), HAS RISEN AT AN ALARMING RATE IN DEVELOPED COUNTRIES DURING THE PAST SEVERAL DECADES. MCS, WHICH EXPRESS THE HIGH-AFFINITY RECEPTOR FOR IGE (FCERI), RELEASE INFLAMMATORY MEDIATORS IN RESPONSE TO ANTIGEN CROSSLINKING OF THEIR SURFACE IGE/FCERI COMPLEX. A G PROTEIN-COUPLED RECEPTOR KNOWN AS MRGPRB2, WHICH IS MAINLY EXPRESSED ON MCS, HAS BEEN DISCOVERED TO BE RESPONSIBLE FOR FACILITATING ANTIBODY-INDEPENDENT MC ACTIVATION, INCLUDING CLINICALLY IMPORTANT DRUG-INDUCED MC ACTIVATION. THE ROLE OF MICROPHTHALMIA-ASSOCIATED TRANSCRIPTION FACTOR (MITF) IN MC DIFFERENTIATION WAS UNCOVERED BY STUDYING MICE WITH VARIOUS SPONTANEOUS MUTATIONS IN THE MITF GENE. HOWEVER, BECAUSE MITF IS IMPORTANT FOR MC DIFFERENTIATION, FEW MATURE MCS DEVELOP IN THE ABSENCE OF MITF. AS A RESULT, THE ROLE OF MITF IN MC LINEAGE COMMITMENT AND FUNCTION IS STILL POORLY UNDERSTOOD. OUR LONG-TERM GOAL IS TO ENHANCE UNDERSTANDING OF THE REGULATION OF MC GENES THAT INFLUENCE ALLERGY SUSCEPTIBILITY AND SEVERITY. THIS APPLICATION INVESTIGATES HOW MITF PROMOTES THE EXPRESSION OF MC-SPECIFIC GENES WHILE SIMULTANEOUSLY SUPPRESSING THE EXPRESSION OF BASOPHIL-SPECIFIC GENES. ADDITIONALLY, IT WILL EVALUATE THE FUNCTIONAL DEFICIENCIES OF MCS WITH AN INDUCIBLY DELETED MITF GENE. OUR PRELIMINARY STUDIES GENERATED MICE WITH A GENETICALLY MODIFIED MITF GENE THAT CAN BE SPECIFICALLY DELETED IN FULLY DIFFERENTIATED MCS. OUR FINDINGS SHOW THAT DELETING THE MITF GENE IN FULLY DIFFERENTIATED BONE MARROW-DERIVED MCS (BMMCS) SUBSTANTIALLY REDUCES FCER1A AND C-KIT EXPRESSION. IMPORTANTLY, MITF-/- BMMCS ALSO HAVE RE-EXPRESSED THE GENE FOR THE BASOPHIL LINEAGE-DETERMINING TRANSCRIPTION FACTOR (TF) CEBPA. IN ADDITION TO THESE OBSERVATIONS, OUR PRELIMINARY RNA-SEQ DATA REVEALED THAT MITF REGULATES THE MRGPRB2 GENE, OPENING A NEW AREA OF INVESTIGATION. BASED ON OUR PRELIMINARY FINDINGS, WE HYPOTHESIZE THAT MITF BOTH PROMOTES MC-SPECIFIC GENE EXPRESSION AND SUPPRESSES BASOPHIL-SPECIFIC GENE EXPRESSION AND THAT MITF REGULATES ANTIBODY-AND DRUG-INDUCED ANAPHYLAXIS. AIM 1 WILL DETERMINE WHETHER MITF MAINTAINS MC-SPECIFIC GENES BY PROMOTING CHROMATIN ACCESSIBILITY AND BY COOPERATING WITH OTHER TFS TO ACTIVATE ENHANCERS THAT REGULATE THE MC-SPECIFIC GENES. AIM 2 WILL DETERMINE HOW MITF SUPPRESSES BASOPHIL CELL FATE IN MCS BY SILENCING THE CEPBA ENHANCERS. AIM 3 OF THIS STUDY WILL EVALUATE THE FUNCTIONAL DEFICIENCIES OF BMMCS WITH AN INDUCIBLY DELETED MITF GENE IN IGE-MEDIATED AND IGG-MEDIATED ANAPHYLAXIS, AS WELL AS ANAPHYLAXIS INDUCED BY THE ANTIBIOTIC CIPROFLOXACIN. RESULTS FROM THESE STUDIES SHOULD ADVANCE OUR UNDERSTANDING OF MC LINEAGE COMMITMENT AND HOW MITF- DEPENDENT MC TRANSCRIPTIONAL PROGRAMING REGULATES ANTIBODY-MEDIATED AND DRUG-INDUCED ANAPHYLAXIS. DECODING ENHANCERS CAN HELP FIND WAYS TO REDUCE THE EXPRESSION LEVELS OF GENES INVOLVED IN MC ACTIVATION AND ANAPHYLAXIS AND THUS CONTROL MC-MEDIATED DISEASES BY PROVIDING TARGETS FOR CRISPR-BASED INTERVENTIONS.

STRUCTURE AND FUNCTION OF EUKARYOTIC PHOSPHATIDYLSERINE DECARBOXYLASE

REPARATIVE FUNCTIONS OF HUMAN AIRSPACE MACROPHAGE SUBSETS - PROJECT SUMMARY FOR LUNG HEALTH TO BE RESTORED AFTER INFLAMMATION, DEAD CELLS MUST BE REMOVED FROM THE AIRSPACES AND THE ALVEOLAR EPITHELIUM MUST BE REPAIRED. AIRSPACE MACROPHAGES (AM) HAVE BEEN IMPLICATED IN CLEARANCE OF APOPTOTIC CELLS, TERMED EFFEROCYTOSIS, AND IN ORCHESTRATING EPITHELIAL REPAIR. WHETHER ALL AM ARE CAPABLE OF THESE FUNCTIONS, OR WHETHER DISTINCT SUBSETS PERFORM SPECIALIZED ROLES FOLLOWING INFLAMMATION IS POORLY UNDERSTOOD. IN PREVIOUS STUDIES, WE LEVERAGED AN ENDOBRONCHIAL LIPOPOLYSACCHARIDE (LPS)-CHALLENGE MODEL OF ACUTE LUNG INFLAMMATION IN HEALTHY ADULTS TO OBTAIN HUMAN AM AND INVESTIGATE THEIR TRANSCRIPTIONAL SIGNATURES. USING SINGLE CELL RNA SEQUENCING, WE DISCOVERED THAT AFTER EXPOSURE TO LPS, THE AIRSPACES ARE DOMINATED BY RECRUITED AM. THESE RECRUITED AM CAN BE DIVIDED INTO TWO DISCRETE SUBSETS BASED, IN PART, ON THEIR EFFEROCYTIC AND REPARATIVE SIGNATURES. HOWEVER, THE CAUSAL MECHANISM BY WHICH EFFEROCYTOSIS IS LINKED TO REPARATIVE PROGRAMING OF AM IN HUMANS HAS NOT BEEN ESTABLISHED. THE PRIMARY OBJECTIVE OF THIS PROPOSAL IS TO IDENTIFY THE UNIQUE FUNCTIONS AND MOLECULAR DRIVERS OF AM SUBSETS DURING RESOLUTION OF INFLAMMATION AND ACTIVE LUNG REPAIR. BASED ON ANALYSIS OF TRANSCRIPTIONAL SIGNATURES AND OUR PRIOR STUDIES, WE POSTULATE THAT A SUBSET OF AM THAT CLOSELY RESEMBLE INTERSTITIAL MACROPHAGES (IMAM) ARE CHARACTERIZED BY TWO UNIQUE FUNCTIONS. FIRST, WE PROPOSE THAT IMAM ARE UNIQUELY PROGRAMED FOR EFFEROCYTOSIS. SECOND, WE HYPOTHESIZE THAT IMAM DRIVE REPAIR OF THE ALVEOLAR EPITHELIUM THROUGH PRODUCTION OF PARACRINE SIGNALING MOLECULES. THE PROPOSED STUDIES WILL INDEPENDENTLY TEST THE DEPENDENCE OF EFFEROCYTOSIS ON DRIVING THESE FUNCTIONS. WE WILL USE PRIMARY AM OBTAINED FROM ENDOBRONCHIAL LPS-CHALLENGE OF RESEARCH PARTICIPANTS AND CUTTING-EDGE ORGANOTYPIC CO-CULTURE SYSTEMS IN THE FOLLOWING THREE SPECIFIC AIMS: 1) TEST THE HYPOTHESIS THAT IMAM ARE SPECIALIZED FOR EFFEROCYTOSIS. 2) TEST THE HYPOTHESIS THAT IMAM PROMOTE ALVEOLAR EPITHELIAL REPAIR. 3) DETERMINE WHETHER EFFEROCYTOSIS IS REQUIRED FOR REPARATIVE PROGRAMING OF HUMAN AM.

RADIATION EXPOSURE SCREENING AND EDUCATION PROGRAM

EXHAUSTION OF AIRWAY PROGENITOR CELLS INDICATES SUSCEPTIBILITY TO COPD

INVESTIGATING THE ROLE OF THE R213G SOD3 POLYMORPHISM IN LUNG DISEASE

NKIP SIGNALING AND FUNCTIONS

OBESITY, INFLAMMATION AND RESPONSE TO THERAPY IN ASTHMA

RADIATION EXPOSURE SCREENING AND EDUCATION PROGRAM

ROLE OF MATRIX METALLOPROTEINASE-3 IN DEPLOYMENT-RELATED PULMONARY FIBROSIS

CELL BASED THERAPY FOR LUNG DISEASE

ANTENATAL DIETARY SUPPLEMENTATION IS A RISK FACTOR FOR INFANT ATOPY THROUGH EPIGE

BIO-REPOSITORY FOR CHRONIC BERYLLIUM DISEASE

STRUCTURE AND FUNCTION OF JMJC HISTONE DEMETHYLASES

LEAD HEALTHY HOME

USE OF THE SRC FAMILY KINASE INHIBITOR SARACATINIB IN THE TREATMENT OF PULMONARY FIBROSIS

UPTAKE OF APOPTOTIC CELLS

METABOLIC REPROGRAMMING OF RECRUITED ALVEOLAR MACROPHAGES BY ARGININE IN ACUTE LUNG INJURY

MODULATING MACROPHAGE-MEDIATED INFLAMMATION IN CYSTIC FIBROSIS

INTERSTITIAL MACROPHAGES IN CIGARETTE SMOKE-INDUCED SMALL AIRWAY REMODELING - PROJECT SUMMARY PROJECT SUMMARY: THIS PROPOSAL DESCRIBES A 5-YEAR RESEARCH TRAINING PROGRAM THAT WILL DEVELOP DR. PATRICK HUME INTO AN INDEPENDENT BASIC AND TRANSLATIONAL PHYSICIAN-SCIENTIST INVESTIGATOR. HIS LONG-TERM CAREER GOAL IS TO ADVANCE THE FIELDS OF COPD AND MACROPHAGE BIOLOGY THROUGH THE ELUCIDATION TARGETS TO BLOCK OR REPAIR THE DEVELOPMENT OF SMALL-AIRWAYS DISEASE IN CIGARETTE SMOKERS. DURING THIS K08 AWARD, DR. HUME WILL GAIN SPECIFIC CAREER DEVELOPMENT TRAINING AND MENTORSHIP CLOSELY ALIGNED WITH AN INNOVATIVE RESEARCH PLAN. HE PROPOSES TO STUDY THE ROLE OF LYVE-1 POSITIVE INTERSTITIAL MACROPHAGES IN THE DEVELOPMENT OF SMALL AIRWAY PATHOLOGY INDUCED BY CHRONIC CIGARETTE SMOKE. GIVEN THE BROAD APPLICABILITY TO COPD, THIS WORK IS DIRECTLY RELEVANT TO THE NHLBI. CANDIDATE: PATRICK HUMAN MD, PHD IS A BOARD-CERTIFIED PULMONARY AND CRITICAL CARE MEDICINE PHYSICIAN- SCIENTIST AT NATIONAL JEWISH HEALTH IN DENVER, CO. HIS RECORD OF BASIC SCIENCE RESEARCH, PH.D. TRAINING AND SCIENTIFIC PUBLICATIONS DEMONSTRATES A FIRM COMMITMENT TO A CAREER AS AN ACADEMIC CLINICIAN-SCIENTIST. TRAINING: THE PROPOSED CAREER DEVELOPMENT PLAN AUGMENTS DR. HUME’S PRIOR MENTORED RESEARCH DURING HIS UNDERGRADUATE, DOCTORAL, MEDICAL RESIDENCY AND FELLOWSHIP TRAINING. HE PROPOSES TO MEET HIS SHORT-TERM OBJECTIVES THROUGH AN INTEGRATED COMBINATION OF INTENSIVE MENTORING BY INTERNATIONALLY RENOWNED EXPERTS IN MACROPHAGE BIOLOGY AND COPD PATHOGENESIS WITH DIDACTIC AND HANDS-ON EXPERIENCES IN (I) DATA ANALYSIS, (II) CELL BIOLOGY, (III) HYALURONIC ACID SIGNALING, (IV) SCIENTIFIC WRITING AND PRESENTATION, AND (V) LABORATORY LEADERSHIP. MENTOR / ENVIRONMENT: DR. HUME HAS CLOSE WORKING RELATIONSHIPS WITH HIGHLY EXPERIENCED MENTORS AND COLLABORATORS WHO CONTRIBUTE EXPERTISE IN MACROPHAGE AND LUNG BIOLOGY (DRS. JANSSEN AND PETRACHE), COPD PATHOGENESIS AND HYALURONIC ACID SIGNALING (DR. PETRACHE), MATRIX METALLOPROTEINASE BIOLOGY AND TISSUE REMODELING (DR. REDENTE), CREATION OF TRANSGENIC MOUSE MODELS (DR. MATSUDA) AND DESIGN-BASED STEREOLOGY (DR. SMITH). THE PROPOSED ACTIVITIES WILL BE BASED AT NATIONAL JEWISH HEALTH, A TOP-RANKED RESEARCH INSTITUTION. RESEARCH PROJECT: THE PRIMARY OBJECTIVE OF THIS PROPOSAL IS TO IDENTIFY THE MECHANISM BY WHICH PERIBRONCHIAL INTERSTITIAL MACROPHAGES (PBIMS) MEDIATE AIRWAY WALL REMODELING AND PATHOGENESIS IN RESPONSE TO CHRONIC CIGARETTE SMOKE EXPOSURE. SPECIFICALLY, OUR STUDIES WILL TEST THE HYPOTHESIS THAT LYVE-1 EXPRESSED ON A SUBSET OF PBIMS BINDS TO HYALURONIC ACID RESULTING IN MATRIX METALLOPROTEINASE 9 (MMP-9) PRODUCTION, LEADING TO AIRWAY WALL REMODELING AND FIBROSIS. THIS WILL BE TESTED IN HUMAN AND MURINE LUNG TISSUE USING QUANTITATIVE MORPHOMETRY AND IMMUNOFLUORESCENCE, FLOW CYTOMETRY, IN VITRO MODELS OF HYALURONIC ACID SIGNALING, AND TRANSGENIC ANIMAL MODELS. IN DOING SO, THE SPECIFIC RELATIONSHIPS BETWEEN LYVE-1+ PBIM RECRUITMENT, MMP-9 PRODUCTION, AND AIRWAY WALL REMODELING WILL BE ELUCIDATED.

PRENATAL PROGRAMMING OF IMMUNE CELLS FOR TISSUE REPAIR AND REMODELING IN ASTHMA - PROJECT SUMMARY/ABSTRACT AIRWAY REMODELING IS A KEY FEATURE OF ASTHMA. MECHANISMS OF AIRWAY REMODELING ARE NOT FULLY UNDERSTOOD. REMODELING AND INFLAMMATION MAY OCCUR INDEPENDENTLY, SUGGESTING MECHANISTIC DIVERGENCE OF THESE TWO FEATURES IN SOME ENDOTYPES OF ASTHMA. AIRWAY REMODELING HAS BEEN DETECTED IN CHILDREN AS YOUNG AS 1-YEAR- OLD, POINTING TO A POSSIBILITY OF AN INBORN DEFECT. INBORN DEFECTS ARE EITHER GENETIC OR A RESULT OF PARENTAL ENVIRONMENTAL EXPOSURES. FOR ASTHMA, THERE ARE STUDIES SUGGESTING BOTH MECHANISMS. USING A MOUSE MODEL, WE DISCOVERED AN INBORN PREDISPOSITION TO AIRWAY REMODELING AND ASTHMA THAT WAS CAUSED BY MATERNAL EXPOSURE TO DIESEL EXHAUST PARTICLES (DEP). SUSCEPTIBILITY TO AIRWAY REMODELING WAS LINKED TO IMPAIRED EMBRYONIC GROWTH AND EMERGENCE OF A REPARATIVE MAST CELL (MC) SUBSET. EMBRYOS AND PUPS OF DEP EXPOSED MOTHERS HAD REDUCED BODY WEIGHT. INBORN IMPAIRMENT OF GROWTH WAS LINKED TO ABNORMAL ACTIVATION OF LUNG MESENCHYMAL CELLS. COMPARED TO LUNGS OF PBS NEONATES, THE LUNG INTERSTITIUM OF DEP NEONATES WAS MORE STRONGLY POSITIVE FOR ACTA2, A MARKER OF WOUND-HEALING MYOFIBROBLASTS, AND AIRWAY SMOOTH MUSCLE WAS THICKENED. MESENCHYMAL CELL ACTIVATION WAS LINKED TO NUMERICAL EXPANSION OF LUNG MCS. POSTNATAL EXPOSURE TO AN ALLERGEN GREATLY EXACERBATED STROMAL PATHOLOGY, LEADING TO EXUBERANT REMODELING OF THE AIRWAYS. DEP- ELICITED AIRWAY REMODELING WAS MC DEPENDENT; MC DEFICIENCY REDUCED REMODELING, AND UNEXPECTEDLY, ENHANCED T2 INFLAMMATION IN THE LUNG. IN SUM, OUR RESULTS SUGGESTED THAT UNDER CONDITIONS OF HARMFUL MATERNAL EXPOSURE, IN AN EFFORT TO RESCUE EMBRYONIC GROWTH, MC RESPONSES SHIFTED FROM PRO-INFLAMMATORY/PRO-ALLERGIC TO ANTI-INFLAMMATORY AND TISSUE-REPARATIVE. OUR SCRNA-SEQ EXPERIMENT FOLLOWED BY FLOW CYTOMETRY AND ADOPTIVE TRANSFER EXPERIMENTS INDICATED THAT THESE NEW FUNCTIONS WERE ATTRIBUTABLE TO A NOVEL MC SUBSET THAT EXPANDED GREATLY IN OFFSPRING OF DEP-EXPOSED MOTHERS, PRODUCED AN ARRAY OF TISSUE-REPARATIVE AND ANTI- INFLAMMATORY FACTORS, AND AFTER BEING ADOPTIVELY TRANSFERRED INTO LUNGS OF MC KO RECIPIENTS, RESTORED PULMONARY EXPRESSION OF REPAIR/REMODELING MARKERS ACTA2 AND VIM. TO CAPTURE THE POSTULATED “RESCUE” FUNCTION OF THIS SUBSET, WE HAVE NAMED IT “ORPHEUS MCS” AFTER THE GREEK HERO WHO ATTEMPTED TO RESCUE HIS WIFE FROM THE UNDERWORLD. WE THEN FOUND THAT ALLERGIC WHEEZING PRESCHOOLERS HAD INCREASED PERCENTAGE OF ORPHEUS MC PROGENITORS IN THE BLOOD, UNDERSCORING THE HUMAN RELEVANCE OF OUR PROJECT. MECHANISTIC STUDIES ON ORPHEUS MC-SPECIFIC RECEPTORS AND THEIR LIGANDS SUGGESTED ROLES OF TWO LIGANDS IN DEVELOPMENTAL PROGRAMMING OF ORPHEUS MCS. TOGETHER, OUR HYPOTHESIS IS THAT ENVIRONMENTALLY-INDUCED INBORN PREDISPOSITION TO ASTHMATIC AIRWAY REMODELING IS IN PART DUE TO EMERGENCE OF THE TISSUE-REPARATIVE ANTI-INFLAMMATORY ORPHEUS MC SUBSET. IN AIM 1, USING OUR MOUSE MODEL, WE WILL ELUCIDATE ROLES OF ORPHEUS MCS IN ASTHMA. IN AIM 2, WE WILL STUDY IMPORTANCE OF TWO IDENTIFIED LIGANDS IN PROGRAMING ORPHEUS MC RESPONSES IN ASTHMA. IN AIM 3, WE WILL STUDY RELEVANCE OF THE ORPHEUS MC LINEAGE FOR HUMAN ASTHMA.

MECHANISM OF T CELL RESISTANCE AGAINST TREG-MEDIATED SUPPRESSION IN ASTHMA

INTEGRATIVE METABOLOMICS OF SARCOIDOSIS DIAGNOSIS AND PROGRESSION - PROJECT SUMMARY/ABSTRACT SARCOIDOSIS IS A SYSTEMIC GRANULOMATOUS DISEASE WHOSE PATHOGENESIS INVOLVES BOTH GENETIC AND ENVIRONMENTAL FACTORS. DIAGNOSIS OF SARCOIDOSIS IS DIFFICULT AND OFTEN DELAYED SINCE THERE IS NO CONFIRMATORY LABORATORY TEST. MANY SARCOIDOSIS PATIENTS DEVELOP PROGRESSIVE PULMONARY DISEASE, WHICH IS ASSOCIATED WITH A HIGHER MORTALITY RATE. HOWEVER, NO RELIABLE BIOMARKERS OF DIAGNOSIS OR PROGNOSIS EXIST TO IDENTIFY PATIENTS WITH DISEASE OR AT RISK OF PROGRESSION. IN ADDITION, SIGNIFICANT DISPARITIES EXIST IN SARCOIDOSIS OUTCOMES, WITH RACE AND SEX DIFFERENCES IN LUNG FUNCTION AND MORTALITY, AND AFRICAN AMERICAN (AA) WOMEN HAVING THE WORST OUTCOMES. THUS, THERE IS A NEED FOR STUDIES FOCUSING ON SARCOIDOSIS DIAGNOSIS AND PROGRESSION, INCLUDING A SEX AND GENDER-DIVERSE POPULATION, WITH LIMITED STUDIES FOCUSING ON GENE EXPRESSION, IN OUR EXCITING PRELIMINARY SARCOIDOSIS METABOLOMICS DATA AT NATIONAL JEWISH HEALTH (NJH). WE FOUND THE ARGININE AVAILABILITY INDEX (AAI, ARGININE/ [ORNITHINE+CITRULLINE]) IS LOWER, AND LINOLEATE IS HIGHER IN SARCOIDOSIS CASES VS. CONTROLS. WE ALSO OBSERVED RACIAL AND SEX DIFFERENCES IN THE METABOLOMIC PROFILES: EUROPEAN AMERICAN SARCOIDOSIS CASES HAVE A MUCH LOWER AAI THAN AAS; FEMALE SARCOIDOSIS CASES HAVE MUCH HIGHER LINOLEATE LEVELS THAN MALES. THESE FINDINGS SUPPORT THE FEASIBILITY OF USING METABOLOMICS TO IDENTIFY POTENTIAL BLOOD SIGNATURES. YET, MOLECULAR SIGNATURES FROM A SINGLE OMICS TYPE ARE GENERALLY UNDERPOWERED AND MAY BE BIASED, GIVEN DISEASE COMPLEXITY. THUS A SYSTEMS BIOLOGY INVESTIGATION USING MULTI-OMICS AND ADVANCED MACHINE LEARNING ALGORITHMS WOULD BE USEFUL TO IDENTIFY MOLECULAR SIGNATURES OF PROGRESSIVE VS. NON-PROGRESSIVE PULMONARY SARCOIDOSIS. SPECIFICALLY, THIS PROPOSAL WILL INCORPORATE SINGLE NUCLEOTIDE POLYMORPHISMS (SNPS) AND GENE EXPRESSION WITH PLASMA METABOLOMICS/LIPIDOMICS TO CONSTRUCT AND VALIDATE PREDICTIVE SARCOIDOSIS DIAGNOSIS AND PROGRESSION MODELS. WE HYPOTHESIZE THAT BLOOD ARGININE AND ARGININE METABOLISM PATHWAY-RELATED SNPS AND GENES ARE ASSOCIATED WITH PULMONARY SARCOIDOSIS RISK AND PROGNOSIS, AND THESE BLOOD SIGNATURES DIFFER BY GENETIC ANCESTRY AND SEX. UTILIZING PRE-EXISTING BLOOD SAMPLES AT NJH AS A DISCOVERY COHORT, WE WILL VALIDATE THE RESULTS WITH PROSPECTIVELY COLLECTED BLOOD, LUNG FUNCTION, AND CHEST IMAGING (VALIDATION COHORT) AT NJH AND MOUNT SINAI HEALTH SYSTEM. WE PROPOSED THE FOLLOWING AIMS: AIM 1: DETERMINE BLOOD DIAGNOSTIC SIGNATURES ASSOCIATED WITH PULMONARY SARCOIDOSIS VS. HEALTHY CONTROLS. AIM 2: DEFINE BASELINE BLOOD PROGNOSTIC SIGNATURES OF PROGRESSIVE PULMONARY SARCOIDOSIS INTEGRATING CLINICAL, SNP, GENE EXPRESSION, AND METABOLOMIC/LIPIDOMIC DATA. AIM 3: IDENTIFY THE LONGITUDINAL BLOOD SIGNATURE METABOLOMIC/INTEGRATIVE OMICS DYNAMICS CONTRIBUTING TO PULMONARY SARCOIDOSIS PROGRESSION. THIS WILL REPRESENT THE LARGEST INTEGRATIVE METABOLOMICS STUDY IN A DIVERSE SARCOIDOSIS POPULATION TO DATE, ENABLING THE IDENTIFICATION OF DIAGNOSTIC (AIM 1) AND PROGNOSTIC (AIM 2) BLOOD SIGNATURES WITH LONGITUDINAL ASSESSMENT (AIM 3). ULTIMATELY, THIS STUDY WILL IDENTIFY POTENTIAL CANDIDATES FOR THERAPEUTIC TARGETS AND BIOMARKERS MOTIVATING PREVENTIVE APPROACHES FOR SARCOIDOSIS PROGRESSION IN FUTURE STUDIES.

MANNOSE BINDING LECTIN-DEPENDENT COMPLEMENT ACTIVATION IN EMPHYSEMA

CHILDHOOD ASTHMA RESEARCH AND EDUCATION NETWORK: DENVER

ALVEOLAR TYPE II CELLS IN VITRO: EFFECT OF KGF

STUDY OF ALLERGEN, GENETICS AND ENDOTOXIN (SAGE)

USE OF PHOSPHODIESTERASE INHIBITORS TO EVALUATE THE PATHOBIOLOGY OF CF

MACROPHAGE METABOLISM AFTER TARGET CELL INGESTION REGULATES ANTI-INFLAMMATORY REPROGRAMMING

ADULT MESENCHYMAL PROGENITOR (MPC) REGULATION OF EARLY PATHOGENESIS IN TSC

DEVELOPMENT OF BEHAVIORAL COUPLES TREATMENT FOR SMOKING CESSATION

ROLE OF THE SFTA2/ADGRF5 SIGNALING AXIS IN ALVEOLAR HOMEOSTASIS - PROJECT SUMMARY PULMONARY ALVEOLAR HOMEOSTASIS IS DEPENDENT UPON BALANCED AIRWAY AND TISSUE SURFACTANT POOLS. ALTERATIONS IN ALVEOLAR SURFACTANT POOLS ARE ASSOCIATED WITH INFLAMMATION AND TISSUE DESTRUCTION IN SEVERE LUNG DISEASES INCLUDING ACUTE RESPIRATORY DISTRESS SYNDROME, PULMONARY ALVEOLAR PROTEINOSIS AND GENETIC DISORDERS OF SURFACTANT DYSFUNCTION. WE HAVE PREVIOUSLY SHOWN THAT THE ORPHAN G-PROTEIN COUPLED RECEPTOR ADGRF5 IS A PHYSIOLOGICALLY DOMINANT MOLECULAR PATHWAY WITHIN ALVEOLAR EPITHELIAL CELLS THAT REGULATES ENDOGENOUS ALVEOLAR SURFACTANT POOLS. THE ABILITY TO PHARMACOLOGICALLY MODULATE THE ADGRF5 PATHWAY, BOTH POSITIVELY AND NEGATIVELY, WOULD BE A MAJOR THERAPEUTIC ADVANCE FOR PATIENTS WITH LUNG DISEASES ASSOCIATED WITH PULMONARY SURFACTANT DISORDERS. HOWEVER, THE ENDOGENOUS LIGAND FOR ADGRF5 HAS NOT BEEN IDENTIFIED, REPRESENTING A CRITICAL KNOWLEDGE GAP IN OUR UNDERSTANDING OF THIS KEY SURFACTANT HOMEOSTASIS MECHANISM. PRELIMINARY DATA WITHIN THIS APPLICATION STRONGLY IMPLICATE A NOVEL SECRETED PROTEIN FROM ALVEOLAR TYPE II CELLS, SURFACTANT ASSOCIATED PROTEIN 2 (SFTA2), AS A DRUGGABLE MOLECULAR TARGET TO MODULATE ENDOGENOUS ALVEOLAR SURFACTANT POOLS. WE DEMONSTRATE THAT SFTA2 IS ABUNDANTLY EXPRESSED IN HUMAN AND MOUSE LUNG, AND PULMONARY EPITHELIUM-SPECIFIC DELETION OF SFTA2 IN MICE RESULTS IN EARLY AND PROGRESSIVE ACCUMULATION OF ALVEOLAR SURFACTANT. OUR CENTRAL HYPOTHESIS IS THAT SFTA2 REGULATES ALVEOLAR SURFACTANT LEVELS VIA BINDING TO THE EXTRACELLULAR DOMAIN OF ADGRF5 TO DECREASE SURFACTANT SECRETION AND/OR INCREASE SURFACTANT UPTAKE IN AT2 CELLS. THE GOAL OF THIS PROJECT IS TO SYSTEMATICALLY DEFINE THE SIGNALING PATHWAYS BY WHICH SFTA2 INTERACTS WITH ADGRF5 TO CONTROL SURFACTANT HOMEOSTASIS USING CELL-BASED ASSAYS AND MOUSE GENETICS THROUGH COMPLETION OF THREE INTEGRATED AIMS. IN THE FIRST AIM, WE WILL TEST THE HYPOTHESIS TEST THE HYPOTHESIS THAT SFTA2 REGULATES ALVEOLAR SURFACTANT LEVELS VIA BINDING AND ACTIVATION OF ADGRF5 SIGNALING IN AT2 CELLS. IN THE SECOND AIM WE WILL TEST THE HYPOTHESIS THAT TRANSIENT AMPLIFICATION OF ALVEOLAR SURFACTANT LEVELS VIA INHIBITION OF THE SFTA2/ADGRF5 SIGNALING AXIS IS SUFFICIENT TO PREVENT AND/OR REVERSE ACUTE LUNG INJURY. IN THE THIRD AIM WE WILL DEFINE THE ROLE OF GLYCOSYLATION IN THE TRAFFICKING AND FUNCTION OF SFTA2 IN HOMEOSTASIS AND IN THE CONTEXT OF ACUTE LUNG INJURY. SUCCESSFUL COMPLETION OF THESE SPECIFIC AIMS WILL DELINEATE THE SIGNALING PATHWAYS BY WHICH SFTA2/ADGRF5 SENSES AND REGULATES SURFACTANT AND ALVEOLAR HOMEOSTASIS. THE LONG-TERM GOAL OF THIS WORK IS TO IDENTIFY POTENTIAL THERAPEUTIC TARGETS, INCLUDING SFTA2 ITSELF, THAT WILL PERMIT MODULATION OF ENDOGENOUS SURFACTANT LEVELS TO TREAT HUMAN LUNG DISEASES ASSOCIATED WITH SURFACTANT DYSFUNCTION INCLUDING ACUTE LUNG INJURY (ALI) AND ACUTE RESPIRATORY DISTRESS SYNDROME (ARDS).

THE SOCIAL ENVIRONMENT, PSYCHOLOGICAL DISTRESS, AND CLINICAL OUTCOMES IN COPD

THE ROLE OF TAPERING PACE AND SELECTED TRAITS ON HYPNOTIC DISCONTINUATION

TRANSCRIPTIONAL RESPONSES TO WILDFIRE POLLUTION IN AIRWAY EPITHELIAL CELLS IDENTIFY GENETIC RISK FACTORS AND MECHANISMS OF ASTHMA EXACERBATIONS - PROJECT SUMMARY/ABSTRACT INCREASED LEVELS OF AIR POLLUTION FROM WILDFIRES ARE DIRECTLY ASSOCIATED WITH ASTHMA EXACERBATIONS BUT THE MECHANISMS MEDIATING THIS ASSOCIATION ARE NOT UNDERSTOOD. FURTHERMORE, THE SEVERITY OF ASTHMA SYMPTOMS THAT RESULT FROM WILDFIRE EXPOSURE VARY CONSIDERABLY BY INDIVIDUAL. WE SEEK TO IDENTIFY TRANSCRIPTIONAL REGULATORY ELEMENTS (TRES) THAT CONNECT WILDFIRE PARTICULATE EXPOSURES WITH GENETIC VARIANTS ASSOCIATED WITH ASTHMA TO IDENTIFY SUSCEPTIBILITY FEATURES AND MECHANISMS THAT GOVERN ASTHMA EXACERBATIONS. WE WILL EMPLOY WOOD SMOKE PARTICLE (WSP) EXPOSURE IN PRIMARY AIRWAY EPITHELIAL CELLS CULTURED AT AIR- LIQUID INTERFACE TO MODEL THE INTERACTION BETWEEN THE RESPIRATORY EPITHELIUM AND FINE PARTICULATES RELEASED BY WILDFIRES. WE WILL ASSAY CHROMATIN ACCESSIBILITY AND NASCENT TRANSCRIPTION USING NEXT- GENERATION SEQUENCING TECHNIQUES TO IDENTIFY TRES REGULATED BY WSP. THEN, USING A PERMUTATION- BASED BIOINFORMATIC APPROACH, WE WILL INTERSECT THE GENOMIC COORDINATES OF THESE TRES WITH GENOMIC SUSCEPTIBILITY LOCI FOR ASTHMA. WE WILL THEN CHARACTERIZE THE TRANSCRIPTIONAL FUNCTION OF SINGLE NUCLEOTIDE POLYMORPHISMS (SNPS) THAT RESULT FROM THIS INTERSECTION. WE WILL TEST TRE FUNCTION IN THE CONTEXT OF WSP EXPOSURE WITH AND WITHOUT THE SNPS IN PLASMID-DERIVED LUCIFERASE REPORTERS AND IN GENOMIC DNA, USING THE CLUSTERED REGULARLY INTERSPACED SHORT PALINDROMIC REPEATS (CRISPR) – CAS9 TOOL TO INTRODUCE THE VARIANT ALLELES. FINALLY, TO ASSAY THE PHYSIOLOGIC SIGNIFICANCE OF THESE TRE-SNP OVERLAPS, WE WILL TEST THE ROLE OF THE RS258760 SNP, WHICH INHABITS A TRE UNDER REGULATION BY WSP AND CONTROLS EXPRESSION OF THE GLUCOCORTICOID RECEPTOR, IN MEDIATING INTERLEUKIN-8 SECRETION BY EXPOSING AIRWAY EPITHELIAL CELLS FROM DONORS WITH THE MAJOR AND MINOR ALLELES TO WSP AND DEXAMETHASONE. THIS PROJECT WILL IDENTIFY AND CHARACTERIZE GENOMIC FEATURES THAT CONNECT AIR POLLUTION WITH ASTHMA EXACERBATIONS. THESE LOCI AND THE GENES THEY REGULATE WILL SERVE AS CANDIDATES FOR PHARMACOLOGIC THERAPIES TO MITIGATE ASTHMA SYMPTOMS ASSOCIATED WITH WILDFIRE AIR POLLUTION. FURTHERMORE, THIS PROJECT DEVELOPS A METHOD TO INTEGRATE MULTIPLE MODELS OF DISEASE IN A GENOMIC CONTEXT. THIS STRATEGY MAY BE USED TO PROPOSE NOVEL TREATMENTS FOR ASTHMA.

ENDOTHELIAL MECHANISMS OF IMPAIRED LUNG GAS EXCHANGE BY HIV

SLEEP DEPRESSION, AND PSYCHOSOCIAL RISK FACTORS IN CAREGIVERS

DIFFERENTIATION OF TH2 CYTOKINE-PRODUCING INNATE CELLS

PULMONARY REHABILITATION FOR IDIOPATHIC PULMONARY FIBROSIS

POLYMORPHISMS RELATED TO EFFECTS OF PARTICULATE AIR POLLUTION IN URBAN CHILDHOOD

CD8 MEDIATED APOPTOSIS DURING T CELL DEVELOPMENT

IMMUNE RECOGNITION AND TOLERANCE OF ANTIBODY DIVERSITY

LUNG EPITHELIAL CELL SURVIVAL SIGNALING IN OZONE IN CYSTIC FIBROSIS AND NORMALS

BACTERIA IN LOWER AIRWAYS AND AIRWAY INFLAMMATION IN CHILDREN WITH ASTHMA

ROLE OF BETA-CATENIN IN EPITHELIAL REPAIR IN ACUTE LUNG INJURY

DEVELOPMENT OF A MORPHOMETRIC APPROACH TO QUANTIFICATION OF SMALL AIRWAYS DISEASE AND A PARTICULATE MATTER EXPOSURE PROFILE IN LUNG BIOPSIES OF DEPLO

SUPERCOMPUTER LINUX CLUSTER FOR GENOMICS AND PROTEOMICS

GENETIC EPIDEMIOLOGY OF NONTUBERCULOUS MYCOBACTERIA FOR PREDICTING HOST SPECIFICITY AND LUNG DISEASE OUTCOMES

EFFICACY OF ACTIVATED PROTEIN C IN VENTILATOR ASSOCIATED LUNG INJURY

ICYT REFLECTION HIGH SPEED CELL SORTER

RNA EDITING CONTROLS PULMONARY ENDOTHELIAL PATHOPHENOTYPES IN PULMONARY HYPERTENSION - ABSTRACT THE PRIMARY OBJECTIVE OF THIS PROPOSAL IS TO GENERATE A SCIENTIFIC DISCOVERY PROGRAM IN PULMONARY VASCULAR DISEASE TO ENSURE A ROBUST PATHWAY FOR DR. WOODCOCK TO DEVELOP INTO AN INDEPENDENT RESEARCH SCIENTIST. THIS OBJECTIVE WILL BE ACCOMPLISHED THROUGH A COMBINATION OF ACTIVE MENTORSHIP, DIDACTIC TRAINING, ENRICHMENT ACTIVITIES, AND RESEARCH. THE CORE OF THE RESEARCH FOCUSES ON THE ROLE OF RNA EDITING, A POST-TRANSCRIPTIONAL REGULATION MECHANISM, IN PULMONARY HYPERTENSION. PULMONARY HYPERTENSION (PH) IS A LETHAL DISEASE WITHOUT A CURE. EARLY APOPTOSIS IN PULMONARY ARTERY ENDOTHELIAL CELLS (PAECS) IS A KEY TRIGGER FOR THE DEVELOPMENT OF PH. HOWEVER, THERE IS A CRITICAL KNOWLEDGE GAP REGARDING THE MECHANISMS OR KEY FACTORS CONTROLLING PAEC PATHOPHENOTYPES THAT DRIVE THIS DEADLY DISEASE, AND NO EFFECTIVE THERAPEUTICS EXIST FOR PH THAT TARGET THIS KEY CELL TYPE. ADENOSINE DEAMINASE ACTING ON RNA 1 (ADAR1) IS A DOUBLE-STRANDED RNA EDITING ENZYME THAT CONVERTS ADENOSINE TO INOSINE (A-TO-I) IN GENOME-ENCODED RNA TRANSCRIPTS AND PLAYS A VITAL ROLE IN GENE REGULATION AND CARDIAC FUNCTION. HOWEVER, THE ROLES OF ADAR1-MEDIATED RNA EDITING IN THE REGULATION OF PULMONARY VASCULAR FUNCTION ARE UNKNOWN. OUR PRELIMINARY DATA INDICATE THAT ADAR1 PLAYS A CRUCIAL ROLE IN THE REGULATION OF PAEC SURVIVAL AND THE CYTOSOLIC INNATE IMMUNITY RESPONSE. TO IDENTIFY ENRICHED GENES IN PAECS THAT ARE MODIFIED AND REGULATED BY ADAR1 RNA EDITING ACTIVITY, WE RECENTLY PERFORMED IN VITRO RNA SEQUENCING IN ADAR1 KNOCKDOWN PAECS. FROM THIS ANALYSIS, WE FOUND THAT THE CIRCADIAN GENE NOCTURNIN (NOCT) CONTAINS TWO ACTIVE ADAR1 RNA EDITING SITES. NON-EDITED NOCT TRANSCRIPT IS STABLE AND ACCUMULATES WITHIN PAECS LACKING ADAR1 RNA EDITING. FORCED EXPRESSION OF NOCT UPREGULATED KEY INTERFERON TRANSCRIPTIONAL FACTOR IRF7 AND PROMOTED APOPTOSIS IN PAECS. NOCT KNOCKOUT (KO) MICE MITIGATED PH INDUCED BY HYPOXIA AND IL6 EXPOSURE. LASTLY, SILENCED NOCT RESTORED PAEC APOPTOSIS AND IMMUNITY RESPONSE TRIGGERED BY ADAR1 DEFICIENCY. IN OUR PILOT STUDIES, WE EXPLORED KEY ELEMENTS OF THIS CONCEPT BY SHOWING THAT ADAR1 RNA EDITING CAN CONTROL NOCT EXPRESSION TO REGULATE PAEC FUNCTIONS AND PH. GIVEN THESE FINDINGS, WE HYPOTHESIZE THAT ADAR1 RNA EDITING DEFICIENCY PROMOTES PH DEVELOPMENT DUE TO INDUCTION OF PULMONARY ARTERY ENDOTHELIAL CELL APOPTOSIS VIA ABERRANT NOCT- DRIVEN INNATE IMMUNITY ACTIVATION. THIS HYPOTHESIS WILL BE ADDRESSED IN THE EXPERIMENTS OF THE FOLLOWING SPECIFIC AIMS: (1) TO DETERMINE THE ROLE OF NOCT IN THE INNATE IMMUNITY-DEPENDENT PAEC SURVIVAL RESPONSE; (2) TO DETERMINE THE EFFECT OF ADAR1-SPECIFIC RNA EDITING ON NOCT EXPRESSION AND FUNCTION IN PAECS; AND (3) TO DEFINE THE IMPACT OF ENDOTHELIAL ADAR1 DEFICIENCY ON NOCT-IFN SIGNALING IN PH IN VIVO. THIS NOVEL MECHANISM MAY HAVE IMPORTANT IMPLICATIONS FOR HOW RNA EDITING REGULATES PA ENDOTHELIAL FUNCTIONS RELATED TO PH PATHOGENESIS, AND ITS ELUCIDATION WILL ADVANCE OUR UNDERSTANDING OF PH DEVELOPMENT AND ULTIMATELY LEAD TO NOVEL STRATEGIES FOR DEVELOPING EFFECTIVE THERAPEUTICS TO TREAT PH. THIS PROJECT WILL PROVIDE ME WITH ADVANCED EXPERTISE IN RNA BIOLOGY AND PULMONARY PHYSIOLOGY THAT WILL STRENGTHEN MY DEVELOPMENT INTO AN INDEPENDENT INVESTIGATOR.

GENOMIC ANALYSIS OF EPITHELIAL GROWTH AND REPAIR MECHANISMS IN ASPIRIN EXACERBATED RESPIRATORY DISEASE

UPGRADED EQUIPMENT AND EXPANSION OF CAGE WASH FACILITIES.

NATURALLY OCCURRING T REGULATORY CELLS CONTROL AIRWAY HYPERRESPONSIVENESS