Ada Forsyth Institute Inc

BIOMARKERS OF PERIODONTAL DISEASE PROGRESSION

SPATIAL ORGANIZATION OF THE ORAL MICROBIOME

A FOUNDATION FOR THE ORAL MICROBIOME AND METAGENOME

GENETIC REGULATORY NETWORK IN CRANIOFACIAL DEVELOPMENT

A FOUNDATION FOR THE ORAL MICROBIOME AND METAGENOME

CULTIVATION, NATURE, ECOLOGY AND PATHOGENICITY OF THE UNCULTIVABLE ORAL MICROBIOM

DOMESTICATION AND CHARACTERIZATION OF TM7-THE MOST ELUSIVE ORAL PHYLUM

MECHANISMS OF RESOLVIN E1 IN PERIODONTAL REGENERATION

MAKING A QUANTUM LEAP IN PLAQUE RESEARCH WITH MODERN SCIENCES

PREVENTING DENTAL CARIES THROUGH TARGETED TREATMENT OF ACID-PRODUCING BACTERIA - ABSTRACT: IN TODAY'S MICROBIOME ERA, IT IS WELL-RECOGNIZED THAT DENTAL CARIES, ONE OF THE MOST PREVALENT AND COSTLY CHRONIC INFECTIOUS DISEASES WORLD-WIDE, RESULTS FROM DYSBIOSIS OF THE ORAL MICROBIOTA AND THE ORAL ENVIRONMENTAL CHANGES THAT CAUSE TOOTH DAMAGE. SPECIFICALLY, FREQUENT INTAKE OF FERMENTABLE CARBOHYDRATES PROMOTES A PROGRESSIVE SHIFT IN MICROBIAL COMPOSITION TOWARD ACIDOGENIC AND ACID-TOLERANT SPECIES. THE CONTINUAL ACID- INDUCED DEMINERALIZATION EVENTUALLY OVERCOMES THE BUFFERING CAPACITY AND ANTI-MICROBIAL PROPERTIES OF SALIVA, LEADING TO IRREVERSIBLE TOOTH DESTRUCTION. THE GOAL OF THIS PROPOSED RESEARCH IS TO PREVENT DENTAL CARIES THROUGH TARGETED TREATMENT OF ACID-PRODUCING BACTERIA (T-TAB). T-TAB WILL PROMOTE A HEALTHY MICROBIAL COMMUNITY THAT IS VITAL FOR MODULATING PH AND PREVENTING ACID-INDUCED TEETH DAMAGE. THE T-TAB WILL BE ACHIEVED BY SELECTIVELY INHIBITING THE GROWTH OF CARIOGENIC BACTERIA THROUGH ENHANCED ANTIMICROBIAL (AM) EFFICACY IN RESPONSE TO THE ACCELERATED ACID PRODUCTION BY THESE BACTERIA IN COMPARISON TO COMMENSAL SPECIES. WE PROPOSE FOUR SPECIFIC AIMS TO DEVELOP, IDENTIFY AND ASSESS EFFECTIVE T-TAB CANDIDATES. IN SPECIFIC AIM 1, WE WILL SYNTHESIZE AND CHARACTERIZE SIX NEW PH-SENSITIVE QUATERNARY PYRIDINIUM SALTS (PH-QPSS). WE EXPECT TO IDENTIFY COMPOUNDS OR COMBINATIONS OF COMPOUNDS THAT PROVIDE T-TAB IN AQUEOUS MIXTURES. WE WILL ENHANCE OUR UNDERSTANDING OF THE CHEMICAL STRUCTURE/AM EFFICACY RELATIONSHIP AND OPTIMIZE THE AM EFFICACY AND SOLUBILITY OF PH-QPS(S) TO OBTAIN SAFE AND EFFECTIVE T-TAB TREATMENTS. IN SPECIFIC AIM 2, WE WILL TRANSFORM A CLINICALLY TESTED AM AGENT, CHLORHEXIDINE (CHX), INTO A T-TAB AGENT WHICH PROVIDES PH-RESPONSIVE AM EFFICACY. WE WILL ACHIEVE ACID ENHANCED CHX RELEASE THROUGH ENCAPSULATED CHX IN QPS-FUNCTIONALIZED MESOPOROUS SILICA NANOPARTICLES. WE WILL ALSO IDENTIFY THE SYNERGISTIC PH-AM-E INDUCED BY INTERACTIONS OF CHX AND PH-QPSS. IN SPECIFIC AIM 3, WE WILL ASSESS AND COMPARE THE T-TAB EFFICACY OF LEAD CANDIDATES FROM AIM 1 AND AIM 2 BY EMPLOYING A MULTISPECIES BIOFILM MODEL THAT SIMULATES HUMAN ORAL MICROBIAL COMMUNITY (NAMED O-MIX). THE T-TAB EFFICACY WILL BE ASSESSED IN THE PRESENCE AND ABSENCE OF SUCROSE—THE CARIOGENIC DIETARY CARBOHYDRATE. STRATEGY WILL ENTAIL EVALUATING BIOMASS, ANALYZING MICROBIAL PROFILES AND DETERMINING ENVIRONMENTAL PH. FINALLY, THE MOST EFFECTIVE T-TAB CANDIDATES THAT SUCCESSFULLY INHIBIT THE GROWTH OF CARIOGENIC ACID-PRODUCING BACTERIA WITHOUT AFFECTING THE FUNCTIONS OF COMMENSAL SPECIES WILL BE FURTHER ASSESSED IN SPECIFIC AIM 4 IN VITRO USING A MICROBIAL-CARIES MODEL ON HUMAN ENAMEL AND IN VIVO EMPLOYING A WELL-DEVELOPED MOUSE CARIES MODEL. SUCCESSFUL COMPLETION OF THE PROPOSED AIMS WILL PROVIDE NEW MATERIALS FOR ORAL RINSE IN DENTAL CLINICS TO PREVENT/TREAT DENTAL CARIES. KNOWLEDGE GAINED FROM THIS STUDY WILL ALSO ADVANCE MATERIAL DEVELOPMENT TO PREVENT INFECTION AND EROSION.

L-ARG AVAILABILITY AFFECTS THE PHYSIOLOGICAL STATE OF PORPHYROMONAS GINGIVALIS

NEUTROPHILS AND PERIODONTITIS IN DIABETES

PHYSIOLOGICAL INTERACTION BETWEEN PROBIOTIC BACTERIA AND PORPHYROMONAS GINGIVALIS

CARIES RESISTANCE MECHANISMS IN HIGH-RISK INDIGENOUS CHILDREN - PROJECT SUMMARY THERE IS A FUNDAMENTAL GAP IN OUR KNOWLEDGE WHETHER NATURAL MECHANISMS PROTECT HIGH-RISK CHILDREN FROM CARIES AND ARISE FROM MICROBIAL INTERACTIONS OF COMMENSAL BACTERIA IN THE ORAL CAVITY, OR FROM THE INTERPLAY OF MICROBIOME AND TOOTH, MEDIATED BY SALIVA. THE LONG-TERM GOAL IS TO PREVENT EARLY CHILDHOOD CARIES (ECC) IN NORTH AMERICAN INDIGENOUS CHILDREN THROUGH PROTECTIVE TREATMENTS THAT MAY ALSO BE APPLICABLE IN THE BROADER POPULATION. THE CENTRAL HYPOTHESIS, BASED ON THE RESEARCH TEAM’S STRONG PRELIMINARY DATA, IS THAT CHILDREN WITH AND WITHOUT ECC DIFFER IN ONE OR MORE KEY DRIVERS: I) MICROBIALLY REDUCED ACIDOGENICITY OF S. MUTANS, OR II) ENAMEL AND DENTIN PROPERTIES, COMPOSITION, OR BIOCHEMICAL FINGERPRINT. THE OBJECTIVE IN THIS APPLICATION IS TO INTEGRATE OBSERVATIONAL AND BASIC SCIENCE, FROM ASSOCIATIONS TO EXPERIMENTS THAT TEST UNDERLYING CARIES PROTECTION MECHANISMS IN CHILDREN WITH HIGH LEVELS OF S. MUTANS. THE STUDY RATIONALE IS BASED ON STRONG EVIDENCE THAT I) ROTHIA SP. ACTIVELY CONTROL S. MUTANS ACIDOGENESIS, AND II) ENAMEL AND DENTIN DIFFER IN COMPOSITION BETWEEN THE TWO GROUPS, WITH LEAD, CADMIUM, AND SULFUR LOWER IN TEETH FROM CARIES-FREE CHILDREN COMPARED WITH THE UNAFFECTED REGION OF CARIES-AFFECTED TEETH. THE RESEARCH TEAM PLANS TO PURSUE THE FOLLOWING THREE SPECIFIC AIMS: AIM 1. TEST WHETHER AND HOW ROTHIA AND/OR OTHER ORAL SPECIES MAY MITIGATE THE CARIOGENIC EFFECTS OF ACIDOGENIC BACTERIA. AIM 2. TEST WHETHER AND HOW TOOTH PROPERTIES MODULATE THE SUSCEPTIBILITY TO ACID DISSOLUTION OF ENAMEL AND DENTIN. AIM 3. TEST HOW TOOTH SUBSTRATE OR SALIVA AFFECT ACIDOGENICITY AND SPATIAL STRUCTURE OF BIOFILMS, AND WHETHER SPATIAL STRUCTURE OF BIOFILMS GROWN FROM EX VIVO DENTAL PLAQUE DIFFERS BETWEEN ECC-AFFECTED AND CF CHILDREN. THE CONTRIBUTION IS EXPECTED TO ACHIEVE HIGH IMPACT BY GOING BEYOND SINGLE-RISK FACTOR STUDIES TO INVESTIGATE CARIES-PROTECTIVE MECHANISMS INVOLVING MICROBIAL GENETICS, BIOFILM ORGANIZATION AND TOOTH COMPOSITION. THE PROPOSED RESEARCH IS INNOVATIVE, BECAUSE WE SHIFT FOCUS TO THE SMALL PERCENTAGE OF INDIGENOUS CHILDREN WITH HIGH LOADS OF S. MUTANS AND WITHOUT CARIES HISTORY, USE STATE-OF-THE ART IMAGING TECHNIQUES OF IN VITRO CONTROLLED BIOFILM GROWTH ON STANDARDIZED ENAMEL CHIPS, AND INTEGRATE MULTIMODAL ANALYSES OF ENAMEL AND DENTIN PROPERTIES, BIOCHEMICAL FINGERPRINT, AND MINERAL COMPOSITION. THIS CONTRIBUTION WILL BE SIGNIFICANT BECAUSE DENTAL CARIES DISPROPORTIONALLY AFFECTS NORTH AMERICAN INDIGENOUS CHILDREN. AFTER SUCCESSFUL COMPLETION OF THIS PROJECT, NEW MECHANISTIC INSIGHTS INTO MOLECULAR INTERACTIONS AND PHYSIOLOGICAL FUNCTIONS OF COMMENSAL ORAL FLORA TO REDUCE ACID PRODUCTION IN CARIOGENIC SPECIES CAN INFORM NEW CARIES PREVENTIVE THERAPEUTIC STRATEGIES.

METAPANGENOMICS OF THE ORAL MICROBIOME - PROJECT SUMMARY/ABSTRACT THE ORAL MICROBIOME IS A COMPLEX MULTISPECIES ECOSYSTEM. DECIPHERING THE INTERACTIONS OF MICROBES WITH EACH OTHER AND THEIR HOST HABITATS IS ESSENTIAL FOR UNDERSTANDING THE ORAL MICROBIOME'S ROLE IN HEALTH AND DISEASE AND FOR DEVELOPING PREVENTIVE AND THERAPEUTIC STRATEGIES. TO SUCCESSFULLY MODULATE THE MICROBIOME WE NEED TO UNDERSTAND THE GENOMIC DRIVERS OF BACTERIAL SITE TROPISMS AND COMMUNITY MEMBERSHIP. MUCH OF THE MOLECULAR DATA NEEDED TO ADDRESS THESE FUNDAMENTAL QUESTIONS ALREADY EXISTS IN PUBLICLY AVAILABLE DATABASES, BUT HAS NOT BEEN ANALYZED WITHIN AN ECOLOGIC FRAMEWORK USING APPROPRIATE BIOINFORMATIC TOOLS AND IN SUFFICIENT DEPTH. WHILE GENOME INFORMATION EXISTS FOR MOST BACTERIAL SPECIES IN THE MOUTH, THERE HAS BEEN NO SYSTEMATIC ANALYSIS OF WHAT GENES ARE CORE OR ACCESSORY FOR THE MAJOR SPECIES – INFORMATION CRITICAL FOR ASSESSING MICROBIAL FUNCTION AND FOR SELECTING STRAINS FOR IN VITRO MODELS. THE SCIENTIFIC PREMISE OF OUR PROPOSAL IS THAT THE COMBINATION OF PANGENOMIC ANALYSIS WITH ORAL METAGENOMIC INFORMATION, HENCE METAPANGENOMICS, WILL TEST THE SITE-SPECIALIST HYPOTHESIS AND PROVIDE UNIQUE, GENE-LEVEL INSIGHTS INTO THE FUNCTION OF THE ORAL MICROBIOME. WE WILL CONSTRUCT PANGENOMES FROM HIGH-QUALITY GENOMES IN THE NATIONAL CENTER FOR BIOTECHNOLOGY INFORMATION (NCBI) TO IDENTIFY CORE AND ACCESSORY GENES OF ORAL SPECIES. WE WILL MAP METAGENOMIC DATA FROM THE HUMAN MICROBIOME PROJECT (HMP), AND OTHER SOURCES, ONTO THE PANGENOMES TO ASSESS WHICH GENOMES ACTUALLY OCCUR IN THE HUMAN MOUTH. ANALYSIS BY ORAL SITE WILL IDENTIFY SITE-TROPISMS AT THE GENOMIC LEVEL, THUS EVALUATING THE SITE- SPECIALIST HYPOTHESIS. WE WILL DETERMINE WHICH GENES ARE UNIQUE TO A SPECIES OR STRAIN OCCUPYING A PARTICULAR ORAL NICHE, AND WE WILL IDENTIFY CANDIDATE GENE DRIVERS OF SITE TROPISM. THE PRODUCTS OF OUR ANALYSIS WILL BE MADE AVAILABLE TO THE ORAL RESEARCH COMMUNITY BY EXPANDING THE HUMAN ORAL MICROBIOME DATABASE (HOMD), DEVELOPED BY DR. DEWHIRST, A PI ON THIS PROPOSAL. PANGENOMES WILL BE MADE AVAILABLE INTERACTIVELY VIA A NEW PLATFORM FOR ANALYSIS AND VISUALIZATION OF OMICS INFORMATION (ANVI'O) CREATED BY DR. EREN, A PI ON THIS PROPOSAL. THE IMPACT OF THE PROJECT IS THAT IT WILL ESTABLISH A GENOMIC, ECOLOGIC FRAMEWORK FOR UNDERSTANDING THE ORAL MICROBIOME. IT WILL PROVIDE A DETAILED, SPECIES, STRAIN- AND GENE-LEVEL ANALYSIS OF THE ORAL MICROBIOME IN THE CONTEXT OF INDIVIDUAL ORAL SITES. IT WILL IDENTIFY KEY GENES FOR FOLLOW-UP MECHANISTIC STUDIES ON SITE TROPISMS, NICHE ADAPTATION, AND PATHOGENESIS. IT WILL IMPROVE AUTHENTICATION AND ERROR-CORRECTION OF KEY BACTERIAL SPECIES AND STRAINS. THE PROJECT BUILDS ON THE EXISTING STRENGTHS OF HOMD, NCBI AND HMP, MAKES AVAILABLE TO RESEARCHERS THE RESULTS OF CUTTING-EDGE GENOMIC ANALYSIS, AND LAYS THE GROUNDWORK FOR PROPELLING ORAL MICROBIOME RESEARCH INTO THE PANGENOMIC AND METAGENOMIC ERA.

FORSYTH POSTDOCTORAL TRAINING IN ORAL HEALTH RESEARCH

ORAL MICROBIAL BIOMARKERS IN PERIODONTAL DISEASE PROGRESSION

TB VACCINE CANDIDATES FOUND IN PATIENTS' BODILY FLUIDS AND MURINE MHC-1 MOLECULE

STEM CELLS FOR CRANIOFACIAL BONE REPAIR AND REGENERATION

VIRTUAL TWIN-POWERED RAPID DEVELOPMENT OF BIOACTIVE MULTIFUNCTIONAL DENTAL RESTORATIVES - SUMMARY THERE IS A GLOBAL NEED TO PRIORITIZE THE DEVELOPMENT OF A NOVEL DENTAL RESTORATION THAT MEETS THE CRITERIA OF BEING LONG-LASTING, EASY TO USE, AND ACCESSIBLE WITHOUT COMPLICATED EQUIPMENT. WE AIM TO REVOLUTIONIZE DENTAL CARE BY DEVELOPING A CUTTING-EDGE PLATFORM FOR THE RAPID AND PRECISE CREATION OF DENTAL RESTORATION MATERIALS. OUR VISION IS TO TAILOR THESE MATERIALS TO INDIVIDUAL NEEDS, ADAPTING TO THE UNIQUE CONDITIONS OF PATIENTS' ORAL ENVIRONMENTS. OUR INNOVATIVE RESTORATIVE SYSTEM WILL INCLUDE SELF-STRENGTHENING ADHESIVES, BIOACTIVE FILLERS THAT RESPOND TO BIOLOGICAL SIGNALS, DURABLE RESINS, AND ADDITIVES THAT PROVIDE NEW FUNCTIONS. DESIGNED TO BE ADAPTABLE, THIS NEW CLASS OF DENTAL MATERIALS WILL BE ABLE TO REPAIR MICROCRACKS AUTONOMOUSLY AND PREVENT CARIOGENIC BIOFILM. MOREOVER, THE EASILY APPLICABLE AND DURABLE RESTORATIVE SYSTEM WILL ADVANCE DIGITAL DENTISTRY TO A NEW LEVEL. OUR NEW APPROACH INTEGRATES A PHYSICS-BASED MODEL OF TESTING AND DATA-BASED SIMULATIONS INTO THE WORKFLOW OF MATERIAL DESIGN AND EVALUATION, THIS WILL DRASTICALLY EXPEDITE THE TRADITIONALLY SLOW AND COSTLY ROUTE OF MATERIAL DEVELOPMENT. OUR ULTIMATE GOAL IS TO INTRODUCE A NOVEL OPERATIONAL PLATFORM IN RESTORATIVE DENTISTRY, ALONG WITH A SUITE OF MULTIFUNCTIONAL MATERIALS TAILORED FOR VARIOUS DENTAL TREATMENTS. OUR DIVERSE TEAM, WITH EXPERTISE SPANNING MATERIALS SCIENCE, ENGINEERING, DENTISTRY, AND COMPUTATIONAL MODELING, IS COMMITTED TO ACHIEVING THIS THROUGH A THREE-PRONGED STRATEGIC APPROACH. AIM 1 IS TO CREATE A SELF-IMPROVING DENTAL ADHESIVE THAT ENHANCES ITS BOND WITH THE TOOTH SUBSTRATE BY FACILITATING MINERALIZATION AT THE INTERFACE. THIS AUTO-ENHANCING UNIVERSAL DENTAL ADHESIVE (AUDA) WILL FILL GAPS BETWEEN THE TOOTH AND ADHESIVE, OFFERING BETTER ADHESION THAN CURRENT PRODUCTS. WE WILL EXPEDITE AUDA DEVELOPMENT USING HYBRID MODELING AND SIMULATION APPROACHES (HMSA), WHICH INCORPORATE DATA-DRIVEN MULTI-PHYSICS SIMULATION TECHNIQUES AND CREATE A VIRTUAL TWIN OF THE PHYSICAL TESTING ENVIRONMENT. THE PERFORMANCE OF AUDA WILL UNDERGO RIGOROUS VALIDATION, INCLUDING TESTS FOR BOND STRENGTH TO EXTRACTED HUMAN TEETH AND DURABILITY UNDER VARIOUS CHALLENGES SIMULATING HUMAN ORAL ENVIRONMENTS. AIM 2 FOCUSES ON CRAFTING BIOACTIVE DENTAL COMPOSITES WITH SELF-HEALING AND ANTIMICROBIAL PROPERTIES, ACHIEVED BY INTEGRATING INNOVATIVE NANOFILLERS. THESE COMPOSITES THAT ARE DESIGNED TO BE COMPATIBLE WITH EXISTING DENTAL ADHESIVES AND AUDA WILL BE VALIDATED FOR MECHANICAL STRENGTH, BACTERIAL INHIBITION, HEALING EFFICIENCY, AND FATIGUE LIFE IMPROVEMENT. AIM 3 IS TO PRODUCE CAD/CAM RESTORATIVES OPTIMIZED FOR DIGITAL DENTISTRY, OFFERING ROBUST, BIOCOMPATIBLE MATERIALS WITH EXCELLENT AESTHETICS AND BONDING STRENGTH TO TEETH. ULTIMATELY, OUR INTEGRATION OF ARTIFICIAL INTELLIGENCE AND A VIRTUAL LAB WITH THE MATERIAL DEVELOPMENT PROCESS IN THE PHYSICAL LAB WILL REVOLUTIONIZE AND INDIVIDUALIZE DENTAL CARE. THIS PLATFORM WILL PROVIDE MATERIALS PRECISELY TAILORED TO MEET DIVERSE PATIENT NEEDS. SUCCESSFUL COMPLETION OF THESE AIMS WILL GENERATE A BATTERY OF NOVEL BIOACTIVE MULTIFUNCTIONAL RESTORATIVE MATERIALS FOR PRECISION PLACEMENT AND PATIENTS' NEEDS. ITS SUCCESS WILL REDEFINE DENTAL RESTORATIVE MATERIALS, USHERING IN AN ERA OF PERSONALIZED CARIES TREATMENT.

IMPACT OF SACCHARIBACTERIA AND THEIR BACTERIAL HOSTS IN PERIODONTAL AND INFLAMMATORY DISEASES - ABSTRACT HUMAN ORAL POLYMICROBIAL COMMUNITIES PLAY A SIGNIFICANT ROLE IN HEALTH AND DISEASE. PERIODONTITIS IS A POLYMICROBIAL DISEASE AND IS AMONG THE MOST COMMON INFECTIONS OF HUMANS. RECENT NEXT-GENERATION SEQUENCING STUDIES REVEAL THAT PERIODONTAL POLYMICROBIAL COMMUNITIES ARE COMPRISED OF A LARGE DIVERSITY OF BACTERIAL SPECIES; HOWEVER, WE HAVE LITTLE TO NO UNDERSTANDING OF THE DISEASE-ROLE OF SOME OF THE KEY MEMBERS. SACCHARIBACTERIA (TM7) IS ONE SUCH BACTERIAL GROUP THAT HAS BEEN LINKED TO MULTIPLE INFLAMMATORY DISEASES, INCLUDING PERIODONTITIS. IN ADDITION, TM7 BELONGS TO A MAJOR BACTERIAL LINEAGE TERMED CANDIDATE PHYLA RADIATION. CANDIDATE PHYLA RADIATION ENCOMPASSES ONE-QUARTER OF ALL BACTERIAL DIVERSITY, YET THEY HAVE BEEN RECALCITRANT TO LABORATORY CULTIVATION. ORAL TM7 BACTERIA WERE THE FIRST TO BE CULTIVATED, OPENING THE DOOR TO MANY INTRIGUING QUESTIONS DUE TO THEIR UNIQUE CHARACTERISTICS. TM7 HAS AN ULTRA-SMALL CELL SIZE (200-500 NM) WITH A HIGHLY REDUCED GENOME LACKING ESSENTIAL BIOLOGICAL FUNCTIONS. MORE IMPORTANTLY, IT IS AN EPIPARASITIC BACTERIA THAT GROWS ON THE SURFACE OF ACTINOBACTERIA (I.E., ACTINOMYCES, PSEUDOPROPIONIBACTERIUM), WHICH IS ANOTHER UNDERSTUDIED BACTERIAL GROUP IN HUMANS. ACTINOMYCES HAS BEEN SHOWN TO INDUCE INFLAMMATORY RESPONSE BOTH IN CULTURED CELLS AND IN ANIMAL MODELS. OUR PRELIMINARY MOUSE PERIODONTAL STUDIES WERE DESIGNED TO INVESTIGATE THE ROLE OF TM7 AND ITS HOST BACTERIA, ACTINOMYCES, IN VIVO. TM7/HOST BACTERIA PAIRS INDUCED LESS INFLAMMATORY BONE LOSS COMPARED TO HOST BACTERIA ALONE, SUGGESTING THAT ORAL ACTINOMYCES AND RELATED BACTERIA CAN BE A PATHOBIONT (OPPORTUNISTIC PATHOGEN), AND THEIR PATHOGENICITY IS REGULATED BY TM7 BACTERIA. THE CURRENT PROPOSAL WILL INVESTIGATE PATHOGENIC GENES IN TM7 HOST BACTERIA AND HOW THEY ARE REGULATED BY TM7 BACTERIA VIA METATRANSCRIPTOMICS. IDENTIFIED GENES WILL BE FURTHER TESTED BY SITE-DIRECTED MUTAGENESIS IN THE HOST BACTERIA (AIM 1). THE PROPOSAL WILL ALSO DETERMINE THE PATHOGENIC AND IMMUNOSTIMULATORY ACTIVITY OF TM7 BACTERIA IN EPITHELIAL AND IMMUNE CELL INTERACTION ASSAY AND WILL IDENTIFY TM7 MACROMOLECULES THAT ARE INVOLVED IN THEIR INTERACTION WITH THE EUKARYOTIC CELLS (AIM 2). LASTLY, THIS STUDY WILL EXPAND TO CULTURING AND CHARACTERIZING ADDITIONAL TAXONOMICALLY DIVERSE TM7 BACTERIA FROM THE PERIODONTAL PATIENTS, EFFECTIVELY CREATING A LIBRARY OF TM7 BACTERIA. THIS WILL ALLOW US TO DETERMINE THE GENERAL IMMUNOLOGICAL BEHAVIOR OF TM7/HOST BACTERIA ACROSS DIFFERENT TM7 SPECIES (AIM 3). ACCOMPLISHMENT OF THE PROPOSAL AIMS WILL IMPROVE THE CURRENTLY LIMITED UNDERSTANDING OF TM7 BACTERIA AND ITS INTERACTION WITH THE HOST BACTERIA, AS WELL AS GENERATE THE FIRST UNDERSTANDING OF TM7'S ROLE IN PERIODONTAL AND INFLAMMATORY DISEASES.

SURFACE ECTODERMAL MECHANISM AND MATERNAL INTERVENTION OF NEURAL TUBE DEFECTS

REGULATION AND FUNCTION OF INTERLEUKIN-7 IN PRIMARY SJOGRENS SYNDROME

ENAMEL MATRIX 3D ORGANIZATION AND MATURATION STAGE ION FLOW

GENOMICS OF GENE REGULATION IN P. GINGIVALIS

OSTEOPONTIN AND INTEGRINS IN INNATE IMMUNE RESPONSES TO POLYMICROBIAL INFECTION

INTERMITTENT FASTING RESTORES SALIVARY GLAND FUNCTION IN SJ?GREN?S SYNDROME - SJÖGREN'S SYNDROME (SS) IS A CHRONIC AUTOIMMUNE DISEASE AFFECTING MILLIONS OF AMERICANS, IN WHICH SALIVARY GLANDS ARE THE PRIMARY TARGET OF AUTOREACTIVE T CELLS, LEADING TO HYPOSALIVATION, THE MAJOR DISEASE HALLMARK. HYPOSALIVATION (DRY MOUTH) CAUSES A VARIETY OF ORAL HEALTH ISSUES AND SEVERELY COMPROMISES QUALITY OF LIFE. SS HAS NO CURE AND CURRENT TREATMENTS ARE PREDOMINANTLY PALLIATIVE. FAILURE OF THERAPIES FOR SS ARE INEXTRICABLY LINKED TO THE INABILITY TO CONTROL AUTOINFLAMMATION; THUS, WE PROPOSE TO INVESTIGATE BOTH SIMULTANEOUSLY. TO ACHIEVE SUSTAINABLE SALIVARY SECRETION, DEVELOPMENT OF APPROACHES TO SIMULTANEOUSLY ENHANCE ENDOGENOUS SALIVARY GLAND REGENERATION AND PROTECT THE GLANDS FROM FURTHER INJURY FROM AUTOIMMUNE INFLAMMATION ARE CRITICALLY NEEDED. OUR PRELIMINARY STUDIES STRONGLY SUGGEST THAT THERE ARE PROTECTIVE ACTIONS PROVIDED BY INTERMITTENT FASTING (IF) IN SS. IN NON- OBESE DIABETIC (NOD) MICE, A WELL-DEFINED SPONTANEOUS MODEL OF SS, IF ENHANCES THE PROLIFERATION OF SALIVARY GLAND STEM CELLS, AND UPREGULATES WNT AND NOTCH SIGNALING AND PEROXISOME PROLIFERATOR- ACTIVATED RECEPTOR-DRIVEN FATTY ACID OXIDATION, WHICH ARE CRITICALLY INVOLVED IN THE EXPANSION AND DIFFERENTIATION OF MULTIPLE STEM CELL TYPES. IT ALSO MITIGATES AUTOREACTIVE T HELPER 1, T HELPER 17 AND CYTOTOXIC T CELL RESPONSES IN THE SALIVARY GLAND-DRAINING LYMPH NODES. THE OBJECTIVE OF THIS PROPOSED PROJECT IS TO DETERMINE THE PREVIOUSLY UNEXPLORED IMPACT OF IF ON THE ACTIVITY OF ENDOGENOUS SALIVARY GLAND STEM/PROGENITOR CELLS AND AUTOIMMUNE INFLAMMATION IN SS TO UNRAVEL THE UNDERLYING MOLECULAR AND CELLULAR MECHANISMS OF IF BENEFITS, WITH THE LONG-TERM GOAL OF DEVELOPING EFFECTIVE AND TARGETED THERAPIES TO FUNDAMENTALLY IMPROVE SALIVARY GLAND FUNCTION. BASED ON THE LITERATURE AND OUR PRELIMINARY RESULTS, WE FORMULATED THE CENTRAL HYPOTHESIS THAT IF EXERTS BENEFICIAL ACTIONS ON SALIVARY GLAND RESTORATION THROUGH PROMOTING ENDOGENOUS SALIVARY GLAND REGENERATION AND DIMINISHING T CELL MEDIATED AUTOIMMUNE INFLAMMATION IN SS. THIS HYPOTHESIS WILL BE TESTED THROUGH THE FOLLOWING SPECIFIC AIMS: IN AIM 1, WE WILL DISSECT THE POTENTIAL MOLECULAR MECHANISMS FOR THE IMPACT OF IF ON THE PROLIFERATION AND DIFFERENTIATION OF SALIVARY GLAND STEM CELLS IN THE NOD MOUSE, A SPONTANEOUS MODEL OF SS; IN AIM 2, WE WILL ASSESS THE IMPACT OF IF ON THE SELF-EXPANSION AND CELLULAR PLASTICITY OF DUCTAL SALIVARY GLAND PROGENITOR CELLS AND ACINAR CELLS, THE FUNCTION AND PLASTICITY OF T CELL SUBSETS, AND THE ATTENUATION OF AUTOIMMUNE INFLAMMATION IN THE SALIVARY TISSUES, USING LINEAGE TRACING MOUSE MODELS AND AN INDUCIBLE MODEL OF SS. SUCCESSFUL COMPLETION OF THIS STUDY WILL PROVIDE NEW TARGETS FOR THE DEVELOPMENT OF EFFECTIVE NEW THERAPEUTICS FOR SS-LIKE EXOCRINOPATHY, AS WELL AS PROVIDE INSIGHT INTO THE TREATMENT OF XEROSTOMIA CAUSED BY RADIATION, MEDICATIONS OR AGING, AND OTHER AUTOIMMUNE DISEASES THAT SHARE SIMILARITIES IN THEIR PATHOGENESIS WITH SS.

AMELIORATION OF IMMUNE CELL-MEDIATED PERIODONTAL DISEASE

A FOUNDATION FOR THE ORAL MICROBIOME AND METAGENOME

INTEGRATED WNT SIGNALING AND MATERNAL INTERVENTION OF NEURAL TUBE DEFECTS - PROJECT SUMMARY/ABSTRACT NEURAL TUBE CLOSURE DEFECTS (NTDS) ARE AMONG THE MOST COMMON AND SEVERE BIRTH DEFECTS. THE LONG-TERM GOAL OF OUR RESEARCH IS TO UNCOVER THE FUNDAMENTAL MECHANISMS OF MAMMALIAN NTDS, WHICH MAY TRANSLATE INTO APPLICATIONS FOR PREVENTING NTDS. THE MOST COMMON AND SIGNIFICANT TYPE OF NTDS IS SPINA BIFIDA THAT OCCURS WHEN THE SPINAL COLUMN DOES NOT CLOSE COMPLETELY AND SPINAL CORD AND/OR SPINAL MENINGES MAY PROTRUDE THROUGH THE BACK. NEWBORNS WITH SPINA BIFIDA ARE VIABLE BUT USUALLY HAVE IMPAIRED BLADDER AND BOWEL FUNCTIONS AND PARALYSIS OF THE LOWER LIMBS, WHICH RESULT IN SERIOUS LONG-TERM MORBIDITY AND DISABILITIES, AND EVEN EARLY DEATH. THE RESEARCH TARGETED AT THE CAUSE AND INTERVENTION OF NTDS, PARTICULARLY, SPINA BIFIDA IN THIS PROJECT IS THUS HIGHLY SIGNIFICANT. THE GENETICALLY MODIFIED MUTANT MOUSE MODELS HAVE PROVIDED SIGNIFICANT CLUES OF HUMAN NTDS. AMONG HUNDREDS OF NTD-ASSOCIATED GENES, WNT FAMILY GENES PLAY KEY ROLES IN NEURAL TUBE CLOSURE AND ABNORMAL WNT SIGNALING MAY CAUSE FOLATE-RESISTANT NTDS IN BOTH ANIMAL MODELS AND HUMANS. CRANIORACHISCHISIS, THE SEVEREST BUT RARE NTD WITH AN ENTIRELY OPEN BRAIN AND SPINE, HAS BEEN FOUND IN THE ANIMAL MODELS OF THE NON-CANONICAL WNT/PLANAR CELL POLARITY (PCP) SIGNALING MUTANTS. THE CANONICAL WNT/SS-CATENIN SIGNALING PATHWAY SHARES SEVERAL COMPONENTS WITH THE PCP SIGNALING PATHWAY AND PLAYS CRUCIAL ROLES IN A WIDE RANGE OF DEVELOPMENTAL PROCESSES AND RELATED DISORDERS. OUR PUBLISHED WORKS DEMONSTRATE INDISPENSABLE ROLES OF WNT/SS-CATENIN SIGNALING IN NEURAL TUBE CLOSURE AND SPINAL NTDS. AT THE CELLULAR LEVEL, WE RECENTLY REVEALED PREVIOUSLY UNDERSTUDIED NON-NEURAL SURFACE ECTODERMAL CELLS WHICH FORM THE MULTI-CELLULAR ROSETTE STRUCTURES, CONVERGENT CELLULAR PROTRUSIONS, AND A UNIQUE F-ACTIN CABLE NETWORK AT THE LEADING FUSION SITE DURING NEURAL TUBE CLOSURE. THESE UNIQUE SURFACE ECTODERMAL STRUCTURES ARE SEVERELY DISRUPTED IN THE SURFACE ECTODERMAL MUTANT MICE WITH FULLY PENETRANT SPINAL NTDS. THIS PROJECT IS DESIGNED TO SYSTEMATICALLY ADDRESS THE TRANSCRIPTOMIC LANDSCAPES AND GENE-REGULATORY NETWORKS IN THE SURFACE ECTODERMAL CELLS DURING NORMAL AND DEFECTIVE NEURAL TUBE CLOSURE USING UNBIASED SINGLE-CELL TRANSCRIPTOMICS IN COMBINATION WITH CONDITIONAL GENE-TARGETING APPROACHES OF THE INTEGRATED WNT SIGNALING AND PCP SIGNALING, WHICH MAY REVEAL NOVEL MECHANISMS UNDERLYING SPINAL NTDS AND PROVIDE A BASIS FOR DEVELOPING NOVEL STRATEGIES TO PREVENT FOLATE-UNTREATABLE NTDS IN HUMAN NEWBORNS THROUGH TRANSCRIPTOMIC MODULATIONS OF THE KEY GENE-REGULATORY NETWORKS.

NONCLASSICAL ?-CATENIN SIGNALING IN ODONTOGENESIS - ABSTRACT THE PRIMARY OBJECTIVE OF THIS APPLICATION IS TO ELUCIDATE THE REGULATORY MECHANISMS UNDERLYING ODONTOGENESIS MEDIATED BY NONCLASSICAL SS-CATENIN SIGNALING. TOOTH AGENESIS IS THE MOST COMMON CONGENITAL DENTAL ABNORMALITY, CHARACTERIZED BY THE ABSENCE OF ONE OR MORE PERMANENT TEETH INCLUDING ANODONTIA, OLIGODONTIA, AND HYPODONTIA. HUMAN GENETIC STUDIES OF NONSYNDROMIC TOOTH AGENESIS HAVE REVEALED APPROXIMATELY 16 CAUSATIVE GENES OF WHICH 6 OF THEM ARE INVOLVED IN THE WNT PATHWAY INDICATING ITS SIGNIFICANCE IN DISEASE PATHOGENESIS. MISSING TEETH ARE CURRENTLY TREATED BY DENTAL IMPLANTS, TOOTH TRANSPLANTS, OR PROSTHODONTIC REPAIRS. HOWEVER, THEY ARE NOT PERMANENT TREATMENTS AND ALSO ARE ASSOCIATED WITH CONSIDERABLE COMPLICATIONS. FOR NEXT-GENERATION THERAPIES, NEW REGENERATIVE APPROACHES TO INTERRUPT TOOTH FORMATION/MAINTENANCE OR DEVELOP AN AUTOLOGOUS TOOTH REPLACEMENT ARE HIGHLY ATTRACTIVE CONCEPTS. THEREFORE, IT IS CRITICAL TO ADVANCE OUR KNOWLEDGE OF ODONTOGENESIS AND ELUCIDATE THE MECHANISMS UNDERLYING RECIPROCAL INTERACTIONS OF DENTAL EPITHELIUM AND MESENCHYME. CANONICAL WNT SIGNALING MEDIATED BY SS-CATENIN HAS BEEN WELL ESTABLISHED TO PLAY AN ESSENTIAL ROLE IN EARLY ODONTOGENESIS. MOUSE GENETIC STUDIES HAVE DEMONSTRATED THE IMPORTANCE OF WNT SIGNALING IN VARIOUS ASPECTS OF DENTAL MEDICINE, E.G. TOOTH DEVELOPMENT, DENTAL PULP, ENAMEL, ODONTOGENESIS, AND AMELOGENESIS. EXPRESSION OF A DEGRADATION-DEFICIENT FORM OF SS-CATENIN CAUSES CONTINUOUS TOOTH GENERATION AND DEVELOPMENT OF SUPERNUMERARY TEETH, FURTHER SUGGESTING THAT TOOTH RENEWAL CAN BE UNLOCKED BY INCREASING THE INTRINSIC LEVEL OF ODONTOGENIC POTENTIAL. SS-CATENIN ACTS AS A MASTER REGULATOR OF THIS INTRINSIC POTENTIAL TO PROMOTE TOOTH FORMATION. HOWEVER, SS-CATENIN IS A MULTIFACED PROTEIN THAT POSSESSES OTHER FUNCTIONS IN ADDITION TO ACTING AS A MASTER REGULATOR FOR TRANSDUCING CANONICAL WNT SIGNALING. PRELIMINARY STUDIES OF OUR NEW GENETIC MODELS ARGUE AGAINST CURRENT KNOWLEDGE AND IMPLICATE THE REQUIREMENT OF NONCLASSICAL SS-CATENIN IN ODONTOGENESIS. FIRST, WE WILL CHARACTERIZE NEW SS-CATENIN MUTANT MICE TO RIGOROUSLY ASSESS THE DUAL FUNCTION OF SS-CATENIN IN THE DEVELOPMENT OF ODONTOGENIC ABILITY, AND DIFFERENTIATION OF ODONTOBLASTS AND AMELOBLASTS. SECOND, WE WILL IDENTIFY THE MASTER REGULATORS ACTING DOWNSTREAM OF SS-CATENIN BY A SINGLE-CELL TRANSCRIPTOMIC APPROACH TO DECIPHER THE GENETIC REGULATORY NETWORK ASSOCIATED WITH THE NONCLASSICAL SIGNALING CASCADE. THE OBJECTIVE OF THIS STUDY HAS GREAT SIGNIFICANCE IN HUMAN HEALTH AND REGENERATIVE DENTAL MEDICINE. ELUCIDATING THE MECHANISM UNDERLYING THE REGULATION OF ODONTOGENESIS PROMISES IMPORTANT INSIGHTS INTO NEXT-GENERATION THERAPY FOR DENTAL RESTORATION.

STUDYING THE PROTECTIVE EFFECTS OF NORMAL ORAL FLORA

PASSIVE IMMUNOTHERAPY FOR PEDIATRIC MUTANS STREPTOCOCCAL INFECTIONS

IDENTIFICATION OF UNCULTIVATED AND UNRECOGNIZED PATHOGENS OF PERIODONTITIS

FORSYTH POSTDOCTORAL TRAINING IN ORAL HEALTH RESEARCH

HOST TRNA-DERIVED SMALL RNAS (TSRNAS) MEDIATE INTERACTIONS BETWEEN HOST AND ORAL MICROBES - HOST MUCOSAL SURFACES ARE HIGHLY SPECIALIZED AND POSSESS A COMPLEX ARRAY OF INNATE AND ADAPTIVE IMMUNITY. THEY PROVIDE THE FIRST LINE OF PROTECTION AGAINST INFECTIOUS AGENTS BY INITIATING PROTECTIVE RESPONSES TO POTENTIAL PATHOGENS. FURTHERMORE, THE SYMBIOTIC RELATIONSHIP OF THE HUNDREDS OF MICROBIAL SPECIES WITH THE HOST REQUIRES A FINE-TUNED RESPONSE AT THE MUCOSAL SURFACE THAT PREVENTS OVERGROWTH OF OPPORTUNISTIC PATHOGENS, BUT SPARING BENEFICIAL MICROBES. WHILE SMALL RNAS (SRNAS) HAVE BEEN REGARDED AS A CLASS OF INTRACELLULAR REGULATORY ELEMENTS, EMERGING STUDIES IN PLANT-PATHOGEN AND HOST-GUT MICROBIOME INTERACTIONS UNCOVER THAT HOSTS CAN EXPLOIT SRNAS, SUCH AS TRANSFER RNA (TRNA)-DERIVED SRNAS (TERMED “TSRNAS”), VIA ENCAPSULATION IN EXOSOMES, AS A NEW MODE OF BACTERIAL GENE MODULATION OR DEFENSE MECHANISM. YET, IT REMAINS TO BE ELUCIDATED WHETHER SUCH SRNA-MEDIATED INTER-KINGDOM GENE MODULATION OR DEFENSE MECHANISM EXIST IN THE CONTEXT OF HOST-ORAL MICROBIOME INTERACTION. TO ADDRESS THIS KNOWLEDGE GAP, THE PRESENT R01 APPLICATION WILL FOCUS ON HOST-DERIVED, EXOSOME-BORNE, SALIVARY TSRNAS, OF WHICH THE BIOLOGICAL FUNCTIONS HAVE REMAINED LARGELY ELUSIVE TO DATE. SPEFICICALLY, WE RECENTLY DEMONSTRATED THAT HUMAN NORMAL ORAL KERATINOCYTE- SPONTANEOUSLY IMMORTALIZED (NOKSI) CELLS RELEASED TWO EXOSOME-BORNE TSRNAS, TSRNA-000794 AND TSRNA- 020498, WHEN CHALLENGED WITH FUSOBACTERIUM NUCLEATUM (FN), A KEY ORAL COMMENSAL AND OPPORTUNISTIC PERIODONTAL PATHOGEN. IMPORTANTLY, THESE TWO TSRNAS CAN BE READILY DETECTED FROM SALIVA IN HEALTHY HUMAN SUBJECTS. INTRIGUINGLY, BOTH TSRNAS EXHIBIT HIGHLY SELECTIVE, FN-TARGETING ANTIMICROBIAL ACTIVITY—DIRECTLY ADDING SYNTHETIC MIMICS OF THESE TWO TSRNAS, BUT NOT SCRAMBLE RNA, TO BACTERIAL CULTURE INHIBITS THE GROWTH OF FN, BUT NOT THAT OF PORPHYROMONAS GINIGIVALIS (PG), A GRAM-NEGATIVE PERIODONTAL PATHOGEN OR STREPTOCOCCUS MITIS (SM), A HEALTH-ASSOCIATED ORAL BACTERIUM. IN PRELIMINARY WORK, WE FURTHER TOOK A MULTI-FACET APPROACH TO IDENTIFY A PUTATIVE RNA-SPECIFIC MEMBRANE TRANSPORTER AS WELL AS CANDIDATE INTRACELLULAR BACTERIAL PROTEIN TARGETS OF FN- TARGETING TSRNAS. BUILDING ON OUR COMPREHENSIVE PRELIMINARY DATA, THE GOAL OF THIS APPLICATION IS TWO-FOLD: (PART I) ACHIEVE MECHANISTIC UNDERSTANDING OF THE CROSS-KINGDOM TRAFFICKING OF HOST-DERIVED FN-TARGETING TSRNAS AND THEIR MODULATING EFFECT ON FN GROWTH DURING NOKSI-FN INTERACTION, THROUGH EXOSOME TRACKING AND IN-DEPTH DISSECTION OF THE TSRNAS TRANSPORTER AND INTRACELLULAR TARGETS IN FN; (PART II) EXPAND OUR WORK TO PROFILE AND COMPARE SALIVARY TSRNAS BETWEEN HEALTHY AND PERIODONTITIS SUBJECTS, WITH A FOCUS ON DEMONSTRATING THE BROAD IMPLICATION OF HOST-GENERATED TSRNAS AS A CONSERVED MECHANISM TO ACHIEVE HOST-MICROBIAL HOMEOSTASIS. THE REALIZATION OF THIS APPLICATION WILL NOT ONLY ADDRESS A FUNDAMENTAL QUESTION BY DEFINING TSRNAS AS A NEW CLASS OF HOST DEFENSE MOLECULES IN MAINTAINING HOST-MICROBIOME HOMEOSTASIS VIA TARGETED MICROBIAL MODULATION, BUT ALSO WILL PAVE THE WAY FOR A NEW THERAPEUTIC STRATEGY AGAINST ORAL DISEASES, CONSIDERING THE ALREADY SUCCESSFUL TRAJECTORY OF NUCLEIC ACID THERAPEUTICS IN RECENT YEARS.

T-REGULATORY CELLS IN PERIODONTITIS

THE FORSYTH CORE CENTER FOR DISCOVERY AT THE HOST-BIOFILM INTERFACE

REGULATORY MECHANISMS CONTROLLING P. GINGIVALIS SURFACE STRUCTURES - PORPHYROMONAS GINGIVALIS (PG) IS AN ENDOGENOUS PATHOGEN, STRONGLY IMPLICATED IN THE ETIOLOGY OF ADULT PERIODONTAL DISEASE. THIS RESEARCH WILL INCREASE OUR UNDERSTANDING OF THE MOLECULAR MECHANISMS THAT CONTROL SYNTHESIS OF PG’S CAPSULE AND MEMBRANE LIPIDS. OUR OVERARCHING MODEL IS THAT THE BIOFILM STATE ACTS AS A RESERVOIR OF BACTERIA, WHILE CAPSULE SYNTHESIS IS LINKED TO A TRANSITION TO VIRULENCE AND DISRUPTION OF HOMEOSTASIS. WE HAVE SHOWN THAT DNABII FAMILY MEMBERS IN PG (HU PG0121 AND HU PG1258) ARE INVOLVED IN CONTROLLING PRODUCTION OF CAPSULE. IN GENERAL, DNABII PROTEINS ARE KNOWN TO BE CRITICAL FOR REGULATION OF CELL METABOLISM, THE RESPONSE TO ENVIRONMENTAL PERTURBATIONS, AND IN CONTROLLING THE TRANSITION TO AND FROM A QUIESCENT STATE. CENTRAL TO OUR STUDIES IS AN ANTISENSE RNA (ASSUGR, FOR ANTISENSE SURFACE GLYCAN REGULATOR) ENCODED AT THE 5’-END OF THE CAPSULE LOCUS (PG0104-PG0121) WITHIN A NOVEL 77BP INVERTED REPEAT (77BPIR) ELEMENT. DELETION OR OVER- EXPRESSION OF ASSUGR ALTERS THE SYNTHESIS OF BOTH K- ANTIGEN CAPSULE AND LPS, AND EXPRESSION IS, IN PART, CONTROLLED BY THE TWO-COMPONENT RESPONSE REGULATOR PG0720. IN ADDITION, WE HAVE DETERMINED THAT SYNTHESIS OF A CERTAIN SUBSET OF SPHINGOLIPIDS IS REQUIRED FOR THE PRESENTATION OF K-ANTIGEN CAPSULE ON THE CELL SURFACE, AND THIS FINDING ALIGNS WITH OUR DISCOVERY OF A MATCHING ASSUGR TARGET SEQUENCE IN A SPHINGOLIPID-SYNTHESIS LOCUS (PG1780-PG1788). OUR WORKING MODEL IS THAT GENES INVOLVED IN K- ANTIGEN CAPSULE AND SL-SYNTHESIS ARE COORDINATELY REGULATED BY ASSUGR. IN THESE STUDIES, WE WILL DETERMINE HOW DNABII PROTEINS ALONG WITH OTHER REGULATORY PROTEINS AND ASSUGR MODULATE THE SYNTHESIS OF CAPSULE, LPS, AND SPHINGOLIPIDS AND WE WILL ELUCIDATE THE LIGAND/SIGNAL THAT ACTIVATES THE TWO-COMPONENT SYSTEM (PG0719-PG0720). OUR OVERALL GOAL IS TO IDENTIFY REGULATORY PATHWAYS THAT CONTROL THE SWITCH FROM A PERSISTENT, SURFACE-ATTACHED STATE TO A VIRULENT STATE CAPABLE OF DISRUPTING MICROBE-HOST HOMEOSTASIS. THE RESEARCH PROPOSED IN THIS APPLICATION IS SIGNIFICANT BECAUSE UNDERSTANDING THE CONTROL OF SURFACE PROPERTY CHANGES IS A VITAL LINK TO UNDERSTANDING THE SWITCH THIS BACTERIUM MAKES TO A VIRULENT PATHOGEN. AS AN OUTCOME OF THESE STUDIES, WE WILL HAVE CHARACTERIZED MECHANISMS THAT CONTROL THE SYNTHESIS OF KEY VIRULENCE DETERMINANTS. THIS INFORMATION WILL LEAD TO A BETTER UNDERSTANDING OF THE REGULATORY NETWORKS THAT EITHER DIRECT P. GINGIVALIS TO BECOME A VIRULENT PATHOGEN OR TO CONTINUE TO LIE LOW AND PERSIST. OUR RESULTS WILL POTENTIALLY LEAD TO THE DEVELOPMENT OF NEW THERAPEUTIC STRATEGIES FOR MODULATING BIOFILM GROWTH AND PERSISTENCE OF THIS ORAL PATHOGEN.

FORSYTH EXPANSION FOR THE CENTER FOR DISCOVERY AT THE HOST-BIOFILM INTERFACE

PHASE 1/2 CLINICAL TRIAL TO ASSESS THE SAFETY AND PRELIMINARY EFFICACY OF LIPOXIN ANALOG BLXA4-MEORAL RINSE FOR THE TREATMENT OF GINGIVITIS

CORE FACILITIES IMPROVEMENT: FORSYTH DATA CENTER RENOVATION

FORSYTH POSTDOCTORAL TRAINING IN ORAL HEALTH RESEARCH

MATRIX PROTEIN REGULATION OF ENAMEL MINERAL FORMATION

HYDROLYSIS-RESISTANT RESIN NETWORKS FOR DURABLE AND MULTIFUNCTIONAL DENTAL RESTORATIONS - SUMMARY: THE SHORT AVERAGE SERVICE LIFE AND THE RISING INCIDENCE OF SECONDARY CARIES CURRENTLY HAMPER THE RELIABILITY OF EXISTING DENTAL COMPOSITE RESTORATIONS. THIS UNDERLINES THE PRESSING NEED FOR INNOVATIVE, LONG-LASTING DENTAL RESTORATIONS COMPOSED OF NOVEL RESIN NETWORKS, MULTIFUNCTIONAL FILLERS AND ADDITIVES, AND ADHESIVE CONSTITUENTS. OUR GOAL IS TO CREATE SUPERIOR RESINS AND RESIN COMPOSITES FOR DENTAL RESTORATIONS. THESE NEW RESTORATIONS WILL SURPASS THE CAPABILITIES OF THE EXTANT METHACRYLATE, ESTER-BASED ANALOGS. OUR STRATEGY CENTERS ON FORMULATING AND TESTING NOVEL RESINS AND RESIN COMPOSITES THAT RESIST HYDROLYSIS AND ENZYMOLYSIS AND POSSESS ANTIMICROBIAL, PROTEIN-REPELLENT, AND THERAPEUTIC ATTRIBUTES, CONSEQUENTLY PRODUCING DENTAL ADHESIVES AND RESIN COMPOSITES WITH PERFORMANCE AND A LIFESPAN EXCEEDING THEIR METHACRYLATE-BASED EQUIVALENTS CURRENTLY BEING USED IN DENTAL CLINICS. WE WILL ACHIEVE OUR GOAL IN THREE SPECIFIC AIMS. AIM 1: WE WILL DEVELOP AND EVALUATE BIOSTABLE HYBRID RESIN NETWORKS THAT MAY WITHHOLD THE HARSH CHALLENGES IN PATIENTS’ ORAL ENVIRONMENTS. BY STRATEGICALLY ALTERNATING THE PACKING OF THE METHACRYLATE UNITS WITH HYDROPHOBIC STYRENE-DERIVATIVE UNITS, WE WILL PROTECT ESTER-GROUPS FROM HYDROLYSIS, ENZYMOLYSIS, AND CARIOGENIC BACTERIA ATTACK. THE NOVEL BIOSTABLE HYBRID RESIN NETWORKS WILL ENCOMPASS AN ACIDIC COMPONENT FOR SELF-ETCH DENTAL ADHESIVES AND MULTIFUNCTIONAL COMPONENTS FOR VARIOUS NEEDS IN DENTAL RESTORATIONS. WE'LL THEN EMPLOY A HIGHLY SENSITIVE METHOD TO QUANTITIVELY ASSESS AND RANK THESE HYBRID RESINS AGAINST TRADITIONAL ESTER-BASED RESINS. AIM 2: WE PLAN TO FORMULATE AND DEVELOP NEW MULTIFUNCTIONAL HYBRID RESINS AND RESIN COMPOSITES. THESE MATERIALS SHOULD MATCH OR BE BETTER THAN THEIR ESTER-BASED COUNTERPARTS REGARDING ANTIMICROBIAL, PROTEIN-REPELLENT, AND THERAPEUTIC ASPECTS. WE HYPOTHESIZE THAT THESE HYBRID RESIN NETWORKS CAN SEAMLESSLY INTEGRATE WITH THE FUNCTIONAL ADDITIVES AND FILLERS PREVIOUSLY DEVISED FOR CONVENTIONAL ESTER-BASED DENTAL RESIN NETWORKS. AIM 3: WE WILL SUBJECT THE NEWLY DEVELOPED RESINS AND COMPOSITES TO RIGOROUS TESTING AS RESTORATIONS (COMBINATIONS OF DENTAL ADHESIVES AND RESIN COMPOSITES) ON EXTRACTED HUMAN TEETH UNDER STATIC AND CYCLIC LOADINGS. WE WILL CONFIRM THEIR LONG-TERM PERFORMANCE IN THE MOST CLINICALLY RELEVANT SCENARIOS BY SIMULATING REAL-WORLD CHALLENGES SUCH AS CLINICAL AGING, THERMAL CYCLING, AND CARIOGENIC BIOFILM EXPOSURE. SUCCESSFUL COMPLETION OF THE PROPOSED AIMS WILL RESULT IN THE PRODUCTION OF MULTIFUNCTIONAL, DURABLE DENTAL RESIN RESTORATIONS ADEPT AT ADDRESSING VARIOUS NEEDS FOR TREATING DENTAL CAVITIES.

THE ROLE OF METASTASIS-ASSOCIATED PROTEASES AND PEPTIDES IN PROSTATE CANCER

MOLECULAR AND PATHOGENIC STUDY OF AN ORAL TM7 STRAIN, THE FIRST CULTIVATED ULTRA-SMALL BACTERIUM

NEURONAL AND BACTERIAL DRIVEN INFLAMMATION OF THE DENTAL PULP IN MOLAR HYPOMINERALIZATION - PROJECT SUMMARY MOLAR HYPOMINERALIZATION (MH) IS A PREVALENT ENAMEL DEFECT AFFECTING 15% OF CHILDREN WORLDWIDE AND IS CHARACTERIZED BY SOFT AND POROUS DEMARCATED OPACITIES ON THE ENAMEL SURFACES OF PRIMARY AND PERMANENT DENTITION. THESE SOFT SPOTS ON ENAMEL HAVE ELEVATED PROTEIN CONTENT, HINDERING RESTORATIVE SUCCESS. THIS DEFECTIVE ENAMEL'S UNIQUE STRUCTURE AND COMPOSITION FACILITATE THE CONTINUOUS TRANSMISSION OF ORAL STIMULI TO THE DENTIN-PULP COMPLEX UNDERNEATH THE ENAMEL. THEREFORE, CHILDREN WITH MH SUFFER GREATLY FROM LONGSTANDING HYPERSENSITIVITY ASSOCIATED WITH AN UNDERLYING PULPAL INFLAMMATION CALLED PULPITIS. HOWEVER, THE MECHANISMS OF BACTERIAL-INDUCED PULPITIS FAIL TO EXPLAIN THE MH-ASSOCIATED PULPITIS, HINDERING EFFECTIVE DIAGNOSIS AND TREATMENT. THIS PROJECT AIMS TO DISSECT THE ROLE OF SENSORY NEURON-DRIVEN AND BACTERIAL-DRIVEN ACTIVATION OF PULPITIS IN MH USING IN VIVO AND EX VIVO APPROACHES BY STUDYING THE INNATE IMMUNE RESPONSES OF THE DENTIN-PULP COMPLEX. FINALLY, WE AIM TO TEST THE EFFECT OF ENAMEL REMINERALIZATION ON BACTERIAL COLONIZATION AND BACTERIAL-DRIVEN IMMUNE RESPONSE OF THE DENTAL PULP ORGAN. WE HYPOTHESIZE THAT CONTINUOUS ACTIVATION OF THE SENSORY NEURONS AND BACTERIAL INFECTION CONTRIBUTE TO THE DEVELOPMENT OF PULPITIS IN MH IN TANDEM, AND TREATMENT OF THE SOFT ENAMEL WILL REDUCE THE PULPAL RESPONSE. TWO SPECIFIC AIMS ARE PROPOSED TO TEST THIS HYPOTHESIS: AIM1) THE ROLE OF SENSORY NEURON ACTIVATION IN PULPAL INFLAMMATION WILL BE EXAMINED USING A HYPOMINERALIZED KLK4-KO MOUSE MODEL AND WT MICE. SENSORY AFFERENTS WILL BE ACTIVATED WITH REPETITIVE NOXIOUS STIMULI, AND THE INNATE IMMUNE RESPONSES IN DENTAL PULP TISSUE WILL BE ASSESSED BY FLOW CYTOMETRY, CYTOKINE ANALYSIS, AND IMMUNOFLUORESCENCE IMAGING. AIM 2.1) THE ROLE OF BACTERIAL INFECTION IN MH-ASSOCIATED PULPAL INFLAMMATION WILL BE INVESTIGATED BY QUANTIFYING BACTERIAL COLONIZATION AND ACTIVITY ON ENAMEL SURFACES AND ASSESSING THE THERAPEUTIC IMPACT OF SILVER DIAMINE FLUORIDE. AIM 2.2) THE BACTERIA-INDUCED PULPAL IMMUNE RESPONSE WILL BE ASSESSED. TOPICAL SDF APPLICATION WILL AIM TO REMINERALIZE THE SOFT ENAMEL, REDUCE BACTERIAL COLONIZATION ON THE SURFACE, AND IMPROVE THE BARRIER FUNCTION OF ENAMEL AGAINST BACTERIAL INFECTION, ULTIMATELY REDUCING INFLAMMATORY RESPONSES. THIS RESEARCH PROJECT WILL SHED LIGHT ON THE MECHANISTIC COMPONENTS OF PULPITIS IN MH PATIENTS AND FACILITATE THE RESEARCHER'S TRANSITION TOWARDS AN INDEPENDENT CLINICIAN-SCIENTIST CAREER TRACK. DURING THIS 5-YEAR K08 MENTORED CAREER DEVELOPMENT AWARD (PA-20-203), DR. KARAASLAN WILL RECEIVE TRAINING AND TEST THE PROPOSED TWO AIMS UNDER A MULTIDISCIPLINARY TEAM OF MENTORS AND EXPERTS IN HARD TISSUE BIOLOGY, MICROBIOLOGY, IMMUNOLOGY, AND PULP BIOLOGY. ULTIMATELY, THE RESULTS OF THIS STUDY WILL GUIDE TRANSLATIONAL AND CLINICAL STUDIES TO DEVELOP TARGETED THERAPEUTICS TO PREVENT AND TREAT PULPITIS IN MH PATIENTS AND THE GENERAL POPULATION.

PHOTODYNAMIC THERAPY FOR ENDODONTIC DISINFECTION

THE ROLE OF LRAP IN ENAMEL MINERAL FORMATION

MICRORNA-MEDIATED MECHANISMS IN TEMPOROMANDIBULAR JOINT HEALTH AND DISEASE - ABSTRACT ORAL HEALTH REFERS TO THE WELL-BEING OF THE MOUTH, INCLUDING THE TEETH, GUMS, AND JOINTS, AND IS ESSENTIAL FOR OROFACIAL FUNCTIONS SUCH AS SPEAKING AND CHEWING. THE TEMPOROMANDIBULAR JOINT (TMJ), CRUCIAL FOR SPEAKING, CHEWING, AND SWALLOWING, HAS A UNIQUE CARTILAGE STRUCTURE THAT SUPPORTS BOTH MOVEMENT AND GROWTH. WHILE TMJ OSTEOARTHRITIS (TMJOA) AFFECTS 8-16% OF THE POPULATION, IT REMAINS SIGNIFICANTLY UNDER-RESEARCHED COMPARED TO KNEE OSTEOARTHRITIS, WHICH IMPACTS 14% OF U.S. ADULTS. TREATING TMJ DISEASES IS PARTICULARLY CHALLENGING DUE TO THE SPECIALIZED NATURE OF THE MANDIBULAR CONDYLAR CARTILAGE (MCC), A MULTI-LAYERED STRUCTURE THAT COMBINES CHARACTERISTICS OF BOTH ARTICULAR AND GROWTH PLATE CARTILAGE. UNDERSTANDING HOW THIS CARTILAGE IS REGULATED, ALONG WITH STRATEGIES TO RESTORE DAMAGED CARTILAGE, IS ESSENTIAL FOR DEVELOPING EFFECTIVE TREATMENTS. THIS PROPOSAL AIMS TO INVESTIGATE THE ROLE OF MICRORNAS, SMALL NON-CODING RNAS THAT REGULATE GENE EXPRESSION, IN MAINTAINING TMJ HEALTH, SPECIFICALLY FOCUSING ON THEIR REGULATION OF FIBROCARTILAGE STEM/PROGENITOR CELLS (FCSCS) AND THE HIPPO SIGNALING PATHWAY. THESE MECHANISMS ARE CRITICAL NOT ONLY FOR THE MAINTENANCE OF HEALTHY CARTILAGE BUT ALSO FOR THE REMODING OF DAMAGED CARTILAGE. BY EXPLORING THESE PROCESSES, THIS STUDY AIMS TO FILL THE GAP IN TMJ RESEARCH, OFFERING POTENTIAL BREAKTHROUGHS IN DEVELOPING PERSONALIZED TREATMENT OPTIONS AND RESTORING CARTILAGE INTEGRITY FOR TMJ DISEASE.

FORSYTH/TUFTS LIMB REGENERATION THROUGH BIOELECTRICITY

CRANIOFACIAL MICROSOMIA AND GENETIC VARIATION IN HEMOSTASIS AND VASCULOGENESIS

COLLABORATIVE RESEARCH: MIM: LEARNING HOW MUCUS SHAPES AND MAINTAINS MICROBIOMES -MUCUS, THE SLIMY GEL THAT LINES ALL WET SURFACES IN OUR BODIES, IS A KEY ECOLOGICAL NICHE FOR MICROBIOMES: IT ACCOMMODATES AN INCREDIBLE 100 TRILLION MICROBES, SOMEHOW SELECTING FOR HELPFUL MICROBES WHILE CONTROLLING PROBLEMATIC PATHOGENS. MOREOVER, IT DOES SO IN DISPARATE ORGANISMS RANGING FROM THE SIMPLEST ANIMALS TO CORALS, SNAILS, FISH, AND FROGS. YET, IT IS STILL NOT KNOWN WHAT ENABLES MICROBIAL COMMUNITIES TO GROW AND FUNCTION SO EFFECTIVELY INSIDE MUCUS. NEW EVIDENCE SUGGESTS THAT THE SUGAR-COATED MOLECULES THAT FORM MUCUS SHAPE MICROBIOMES THROUGH PHYSICAL, CHEMICAL, AND NUTRITIVE INTERACTIONS. THIS HYPOTHESIS WILL BE TESTED BY ADDRESSING THREE QUESTIONS: WHAT PROPERTIES OF MUCUS DO MICROBES CARE ABOUT? HOW ARE MICROBES STRUCTURED IN MUCUS -- WHO IS NEXT TO WHOM AND WHY? AND, WHAT ARE THE PROCESSES BY WHICH MICROBIOMES SELF-ORGANIZE IN MUCUS? THIS WORK COMBINES THE DISTINCT AND COMPLEMENTARY EXPERTISE OF THREE SCIENTISTS SPECIALIZING IN MUCUS BIOCHEMISTRY, MICROBIOME STRUCTURE AND MOLECULAR BIOLOGY, AND THE BIOPHYSICS OF MICROBE-MUCUS INTERACTIONS. UNDERSTANDING HOW MUCUS CONTROLS MICROBIOMES COULD YIELD NEW STRATEGIES FOR PROTECTING HUMANS FROM INFECTIONS, AS WELL AS FOR LEVERAGING BENEFICIAL MICROBES THAT CAN HELP HUMANS AND ANIMALS THRIVE IN OTHER WAYS SUCH AS BY IMPROVING FOOD DIGESTION AND SALT TOLERANCE. THE INVESTIGATORS WILL ALSO DEVELOP NEW CITIZEN SCIENCE INITIATIVES AND DEMONSTRATIONS, INTEGRATE RESEARCH AND EDUCATION, AND ENGAGE STUDENTS AND TEACHERS TO HELP CREATE A DIVERSE COMMUNITY OF RESEARCHERS AND CHANGE THE PERCEPTION OF MUCUS FROM A SLIMY WASTE PRODUCT TO A FASCINATING BIOMATERIAL WITH CRITICAL BIOLOGICAL FUNCTIONS. MUCUS IS A CRITICALLY IMPORTANT HABITAT FOR MICROBES. DESPITE ITS PIVOTAL IMPORTANCE TO HUMAN AND ANIMAL FUNCTIONING, HOWEVER, THE MECHANISMS BY WHICH MUCUS INTERACTS WITH MICROBIOMES ARE NOT UNDERSTOOD. INDEED, MUCUS INTERACTS WITH MICROBES IN TWO SEEMINGLY CONFLICTING WAYS, MAINTAINING A DENSE AND DIVERSE HEALTHY MICROBIOME WHILE SIMULTANEOUSLY CLEARING AND DISARMING HARMFUL MICROBES. THIS RESEARCH TEAM WILL UNRAVEL THIS PUZZLE BY INVESTIGATING THE MECHANISMS UNDERLYING THIS KEY RULE OF LIFE, WITH THE HYPOTHESIS THAT MUCUS SHAPES MICROBIOMES THROUGH PHYSICAL, CHEMICAL, AND NUTRITIVE INTERACTIONS. TO TEST THIS HYPOTHESIS, THE INVESTIGATORS WILL (I) EVALUATE HOW MUCIN GLYCANS--THE CHAINS OF DIFFERENT SUGARS THAT DECORATE MUCIN PROTEINS-- SELECT FOR SPECIFIC MICROBES IN NATURAL COMMUNITIES, (II) IDENTIFY THE INFLUENCE OF DIFFERENT MUCUS COMPONENTS ON THE INTRICATE SPATIAL STRUCTURE OF COMPLEX MICROBIAL COMMUNITIES, AND (III) DETERMINE THE MECHANISMS BY WHICH MUCUS REGULATES MICROBIAL COMMUNITY ASSEMBLY. BY REVEALING THE BIOCHEMICAL AND BIOPHYSICAL MECHANISMS BY WHICH MUCINS INFLUENCES MICROBIAL COMMUNITY STRUCTURE (BOTH TAXONOMIC AND SPATIAL), THIS WORK WILL ELUCIDATE THE ESSENTIAL ROLE PLAYED BY MUCUS AS AN ECOLOGICAL HABITAT THAT SUPPORTS THE GROWTH OF BENEFICIAL COMMENSAL MICROBES WHILE ALSO PREVENTING THE OUTGROWTH OF PATHOGENS. THE RESULTS WILL ALSO BENEFIT SOCIETY BY INFORMING APPLICATIONS THAT SEEK TO ENGINEER MUCUS-INSPIRED MATERIALS TO CONTROL ENVIRONMENTAL AND ORGANISMAL MICROBIOMES. THIS WORK WILL ALSO INSPIRE NEW EDUCATIONAL AND OUTREACH EFFORTS FOR ALL AGES TO IMPROVE PUBLIC AWARENESS OF THE TOPIC OF ?MUCUS AND MICROBIOMES?. THIS PROJECT WAS CO-FUNDED BY THE DIVISION OF MATERIALS RESEARCH IN THE MATHEMATICAL AND PHYSICAL SCIENCES DIRECTORATE, AND BY THE SYMBIOSIS, INFECTION AND IMMUNITY GROUP IN THE DIVISION OF INTEGRATIVE ORGANISMAL SYSTEMS. THIS AWARD REFLECTS NSF'S STATUTORY MISSION AND HAS BEEN DEEMED WORTHY OF SUPPORT THROUGH EVALUATION USING THE FOUNDATION'S INTELLECTUAL MERIT AND BROADER IMPACTS REVIEW CRITERIA.

MATRIX PROTEIN REGULATION OF ENAMEL MINERAL FORMATION

TREATMENT OF PERIODONTITIS AND INFLAMMATORY CHANGES IN DIABETES

MESENCHYNMAL STEM CELLS AS A PROTECTIVE NICHE FOR LATENT M. TUBERCULOSIS

MECHANISM OF LYSYL OXIDASE PROPEPTIDE IN ORAL SQUAMOUS CELL CARCINOMA MICROENVIRONMENTS - ORAL CANCER IS THE 6TH MOST COMMON CANCER, MOSTLY TONGUE ORAL SQUAMOUS CELL CARCINOMA (OSCC). SURGERY AND RADIATION TREATMENTS RESULT IN SEVERE QUALITY OF LIFE ISSUES, TISSUE STIFFNESS AND METASTASIS. LYSYL OXIDASE (LOX) IS ONE OF FIVE PARALOGUES. ALL CATALYZE THE FINAL EXTRACELLULAR ENZYMATIC REACTION REQUIRED FOR COLLAGEN BIOSYNTHESIS. ONLY LOX UNDERGOES BIOSYNTHETIC PROCESSING TO RELEASE THE ACTIVE ENZYME AND THE UNIQUE 18 KDA LYSYL OXIDASE PROPEPTIDE (LOX-PP) THAT INHIBITS TUMOR GROWTH. PRELIMINARY DATA SHOW THAT A POLYMORPHIC POINT MUTATION IN THE LOX-PP REGION OF LOX GENE LEADING TO R TO Q AMINO ACID CHANGE IN LOX-PP IS LINKED TO INCREASED OSCC. WE FURTHER SHOW THAT WILDTYPE LOX-PP CROSS-TALKS WITH MACROPHAGES TO SHIFT PHENOTYPE TO INFLAMMATORY TUMOR SUPPRESSIVE CELLS. IN LESION MICROENVIRONMENTS IN 4NQO INDUCED ORAL CANCER WT MICE EXHIBIT TUMOR SUPPRESSIVE- T CELL MARKERS, WHILE KNOCKIN Q LOX-PP (KI) MICE SHOW TUMOR CELLS. WE PUBLISHED THAT LOX-PP, BUT NOT POLYMORPHIC Q LOX-PP, INHIBITS FGFR SIGNALING, AND THAT LOX-PP CAN BIND DIRECTLY TO FGFR1. IN NON-ORAL CANCERS IMMUNOSUPPRESSIVE MACROPHAGES OVEREXPRESS FGF-2 AND FGFR1 AND FGF-2 REGULATES CONVERSION TO TUMOR SUPPRESSIVE CELLS IN VITRO AND IN VIVO. INTERESTINGLY, LOX ENZYMES FROM ALL 5 PARALOGUES CAN BE UP- REGULATED, INCLUDING IN OSCC, TO PROMOTE FIBROSIS. INTEGRATING THESE FINDINGS, WE PROPOSE THAT IN OSCC MICROENVIRONMENTS, THE POLYMORPHIC Q LOX-PP HAS LOST ITS TUMOR SUPPRESSOR FUNCTION VIA ITS INABILITY TO BLOCK FGFR-DEPENDENT MACROPHAGE PHENOTYPE SWICH TO PRO-INFLAMMATORY TUMOR SUPPRESSIVE WITH CONSEQUENCES ON T CELL ACTIVATION AND OSCC OUTCOMES. TO INVESTIGATE THE HYPOTHESIS THAT LOX-PP REGULATION OF MACROPHAGE PHENOTYPE CONTRIBUTES TO OSCC TUMOR SUPPRESSION THAT IS LOST IN Q LOX-PP WE PROPOSE 3 AIMS. IN AIM 1A WE WILL INVESTIGATE IN TUMOR MICROENVIRONMENTS VS ADJACENT REGIONS MACROPHAGE PHENOTYPES AND LOCAL MODULATIONS OF T CELL AND MESENCHYMAL CELL TYPE AND GENE EXPRESSION CHANGES UTILIZING STATE OF THE ART SPATIAL TRANSCRIPTOMICS TECHNOLOGIES IN WT COMPARED TO KI MICE IN THE 4NQO MODEL OF CANCER. THE 4NQO MODEL MIMICS HUMAN OSCC DEVELOPMENT. PATHWAYS WILL BE VALIDATED IN VITRO AND IN VIVO. IN AIM 1B WE WILL DETERMINE THE RELATIVE ABILITY OF RECOMBINANT WT R LOX-PP VS Q LOX-PP TO BLOCK ORAL CANCER DEVELOPMENT IN MICE. IN AIM 1C WE WILL INVESTIGATE HUMAN CANCER TISSUES FOR THE SAME PATHWAY DIFFERENCES IN CLINICAL SAMPLES FROM ORAL CANCER SUBJECTS WITH WT AND VARIANT LOX GENOTYPE. AIM 2A WILL DETERMINE DIFFERENTIAL EFFECTS AND REGULATORY MECHANISMS OF WT VS VARIANT LOX-PP ON MACROPHAGES IN VITRO. IN AIM 2B WE WILL PERFORM A LOSS OF FUNCTION STUDY OF SECRETED FGF-2 KO MICE THAT MAY HAVE RESCUED OSCC RESPONSE TO 4NQO IN LOX-PP KI MICE. IN AIM 2C WE INVESTIGATE THE ABILITY OF LOCAL WILDTYPE AAV LOX-PP INJECTION IN KI MICE WITH OR WITHOUT A LOX ENZYME INHIBITOR (PXS-5505) TO DETERMINE ADDITIVE OR SYNERGISTIC BENEFITS IN 4NQO INDUCED OSCC. IN AIM 3 WE WILL DETERMINE AFFINITY OF LOX-PP AND Q LOX-PP TO FGFR1 IN VITRO, AND PERFORM STRUCTURE-FUNCTION STUDIES, AND IDENTIFY FUNCTIONAL DIRECT LOX-PP BINDING PARTNERS. DATA WILL PROVIDE FUNCTIONAL TARGETS OF LOX-PP IN ORAL CANCER TUMOR SUPPRESSION.

COMPREHENSIVE TRANSCRIPTOME ANALYSIS FOR PORPHYROMONAS GINGIVALIS

SPECTRAL CONFOCAL IMAGING INSTRUMENTATION FOR FORSYTH ADVANCED MICROSCOPY CORE - PROJECT SUMMARY/ABSTRACT THIS IS A PROPOSAL FOR THE PURCHASE OF A LASER SCANNING CONFOCAL MICROSCOPE WITH FLUORESCENCE SPECTRAL IMAGING, NEAR-INFRARED IMAGING, AND CORRELATIVE MICROSCOPY CAPABILITIES. THE INSTRUMENT REQUESTED IS THE ZEISS LSM 980 EQUIPPED WITH A 34-CHANNEL SPECTRAL DETECTOR, EXCITATION DIODE LASERS AND BEAM SPLITTERS, NEAR INFRARED (NIR) LASER AND DETECTORS, AND A ZEN CONNECT SOFTWARE MODULE TO ENABLE CORRELATIVE MICROSCOPY WORKFLOWS. THE INSTRUMENT IS CRITICAL TO THE RESEARCH PROGRAMS OF THE USERS AND WILL REPLACE A DECADE-OLD INSTRUMENT THAT NO LONGER IS GUARANTEED SUPPORT BY THE MANUFACTURER. IT WILL BE PLACED IN THE ADVANCED MICROSCOPY CORE FACILITY OF THE FORSYTH INSTITUTE, AN INDEPENDENT RESEARCH INSTITUTE IN CAMBRIDGE, MA, AND WILL SUPPORT THE FUNDAMENTAL AND APPLIED RESEARCH OF 12 INVESTIGATORS— INCLUDING 5 MAJOR USERS AND 7 OTHER USERS STUDYING THE HUMAN ORAL MICROBIOME, MICROBE-HOST INTERACTION, CRANIOFACIAL BONE STRUCTURE, TOOTH ENAMEL AND SALIVARY GLAND FUNCTION. SPECIFIC PROJECTS FOR WHICH THE INSTRUMENT IS NEEDED BY ITS MAJOR USERS INCLUDE (1) MECHANISMS UNDERLYING SPATIAL INTERACTIONS IN THE ORAL MICROBIOTA; (2) IMAGING STUDIES OF THE STRUCTURE AND ORGANIZATION OF ORAL BACTERIAL BIOFILMS AT MICROMETER AND SUB-MICROMETER SCALES; (3) IMMUNO-FLUORESCENCE ANALYSIS OF GENE EXPRESSION AND PROTEIN LOCALIZATION TO UNDERSTAND THE MECHANISMS OF BONE DEVELOPMENT, REPAIR, AND REGENERATION; (4) THE INTERACTION OF ULTRASMALL BACTERIA AND THEIR HOSTS ON PERIODONTITIS AND INFLAMMATORY DISEASE; AND (5) CHARACTERIZATION OF THE EPISYMBIONT TM7 WITH ITS HOST BACTERIAL CELL. IN ADDITION TO THE ENUMERATED PROJECTS, THE PURCHASE OF THIS INSTRUMENT WILL ENABLE AS-YET UNFUNDED PILOT PROJECTS TO EXPLORE PROMISING RESEARCH AVENUES FOR ORAL HEALTH AND DISEASE. COLLECTIVELY, THESE RESEARCH PROJECTS SUPPORT THE OVERALL MISSION OF THE FORSYTH INSTITUTE TO IMPROVE BOTH ORAL AND OVERALL HEALTH THROUGH TARGETED BASIC RESEARCH AS WELL AS CLINICAL SERVICE AND OUTREACH. ACCESS TO THIS INSTRUMENT WILL ENHANCE THE GOALS OF THESE RESEARCH PROJECTS BY PROVIDING AN ACCESSIBLE, RELIABLE, AND STATE-OF-THE-ART INSTRUMENT TO ENABLE USERS TO ACQUIRE IMAGES OF THE HIGHEST QUALITY AND BIOMEDICAL IMPACT USING EFFICIENT WORKFLOWS. THE TECHNICAL EXPERTISE OF THE CORE STAFF AND A CAREFULLY CONSTRUCTED MANAGEMENT PLAN WILL PERMIT MAXIMAL USE OF ACCESSIBLE USER TIME ON THE INSTRUMENT AT THE HIGHEST PERFORMANCE LEVEL. STRONG INSTITUTIONAL SUPPORT FROM THE FORSYTH INSTITUTE WILL GUARANTEE OPERATING SUPPORT FOR THE INSTRUMENT AND WILL ENABLE THE PURCHASE OF AN OFFLINE PROCESSING WORKSTATION FOR EFFICIENT HIGH-THROUGHPUT ANALYSIS OF THE LARGE IMAGING DATASETS THAT THESE PROJECTS WILL GENERATE. IN SUM, THE CUTTING-EDGE IMAGING INSTRUMENTATION REQUESTED IN THIS APPLICATION WILL ADVANCE THE INSTITUTE’S RESEARCH PROGRAMS AND NIH- FUNDED BIOMEDICAL RESEARCH.

INTEGRATING SOCIAL, BEHAVIORAL & PREVENTIVE SCIENCES TO REDUCE CHILDREN'S CARIES

CHILDHOOD OBESITY PREVENTION: A MODEL FOR SCHOOL-BASED DENTAL PROGRAMS

INFECTIONS OF SPECIFIC PERIODONTAL MICROBIOTA ARE ASSOCIATED WITH RISK FOR AGE-RELATED MACULAR DEGENERATION

CELLULAR SENESCENCE CONTRIBUTES TO SALIVARY GLAND DYSFUNCTION IN SJÖGREN’S SYNDROME - SJÖGREN’S SYNDROME (SS) IS A CHRONIC AUTOIMMUNE DISEASE THAT AFFECTS MILLIONS OF AMERICANS, IN WHICH SALIVARY GLAND INTEGRITY AND FUNCTION ARE IMPAIRED, LEADING TO DRY MOUTH AS THE MAJOR CLINICAL MANIFESTATION. THE INSUFFICIENT SALIVA PRODUCTION ALSO CAN SEVERELY COMPROMISE MULTIPLE FACETS OF ORAL HEALTH AND OVERALL QUALITY OF LIFE. TO DATE, THERE IS NO KNOWN CURE OR EFFECTIVE TREATMENT FOR SS. PREVIOUS STUDIES ON SS PATIENTS HAVE DEMONSTRATED THE PRESENCE OF CELLULAR SENESCENCE IN THE SALIVARY GLANDS. MOREOVER, THE PROPORTION OF SENESCENT SALIVARY GLAND CELLS, PARTICULARLY THOSE LOCATED IN THE SALIVARY GLAND STEM CELL- RESIDING DUCTAL COMPARTMENTS, STRONGLY CORRELATES WITH THE DISEASE SEVERITY, ACCOMPANIED BY A DECLINE IN THE FUNCTION OF SALIVARY GLAND STEM CELLS. CONSISTENT WITH THIS, OUR PRELIMINARY STUDIES WITH SPONTANEOUS AND INDUCIBLE MOUSE MODELS OF SS REVEALED AN INCREASED NUMBER OF SENESCENT CELLS IN THE SUBMANDIBULAR GLANDS, COUPLED WITH A REDUCED STEMNESS OF SALIVARY GLAND STEM CELLS. NOTABLY, ADMINISTRATION OF A SENOLYTIC DRUG TO NON-OBESE DIABETIC (NOD) MICE, A WELL-DEFINED SPONTANEOUS MODEL OF SS SIGNIFICANTLY REDUCED SALIVARY GLAND SENESCENT CELLS AND IMPROVED HYPOSALIVATION DURING SS PROGRESSION. THE OBJECTIVE OF THIS PROPOSED PROJECT IS TO DETERMINE THE PREVIOUSLY UNEXPLORED IMPACT OF CELLULAR SENESCENCE ON SS-ASSOCIATED SALIVARY GLAND DISORDER AND SALIVARY GLAND STEM CELL ACTIVITY, WITH THE LONG-TERM GOAL OF DEVELOPING EFFECTIVE AND TARGETED INTERVENTIONS FOR THIS DISEASE. BASED ON THE LITERATURE AND OUR PRELIMINARY RESULTS, WE HYPOTHESIZE THAT CELLULAR SENESCENCE PLAYS A DETRIMENTAL ROLE IN SALIVARY GLAND SECRETORY FUNCTION IN SS, BY EXACERBATING SALIVARY GLAND INFLAMMATION AND DIMINISHING SALIVARY GLAND STEM CELL ACTIVITIES. THIS HYPOTHESIS WILL BE TESTED THROUGH THE FOLLOWING SPECIFIC AIMS: IN AIM 1, WE WILL DETERMINE THE IMPACT OF CELLULAR SENESCENCE ON SS-ASSOCIATED XEROSTOMIA AND SIALADENITIS THROUGH TWO DISTINCT APPROACHES, INCLUDING SENOLYTIC DRUG-BASED ELIMINATION OF SENESCENT CELLS IN NOD MICE AND HERPES SIMPLEX VIRUS Ɩ THYMIDINE KINASE-MEDIATED ABLATION OF SENESCENT CELLS IN P16-3MR GENETICALLY ENGINEERED MICE WITH SS INDUCTION; IN AIM 2, WE WILL ELUCIDATE THE IMPACT OF CELLULAR SENESCENCE ON THE EXPANSION AND DIFFERENTIATION OF SALIVARY GLAND STEM CELLS AND COMPREHENSIVELY CHARACTERIZE THE TRANSCRIPTOMIC PROFILE OF THESE CELLS IN BOTH SS MOUSE MODELS. SUCCESSFUL COMPLETION OF THIS STUDY WILL PROVIDE A CRITICAL AND RATIONAL FOUNDATION FOR THE DEVELOPMENT OF EFFECTIVE THERAPEUTIC STRATEGIES FOR SS- ASSOCIATED EXOCRINOPATHY AND SHED LIGHT ON THE TREATMENT OF XEROSTOMIA CAUSED BY OTHER FACTORS SUCH AS MEDICATIONS OR AGING, AS WELL AS OTHER AUTOIMMUNE DISEASES THAT SHARE SIMILAR PATHOGENIC MECHANISMS WITH SS.

CONTROLLING AUTOIMMUNE INFLAMMATION AND PROMOTING SALIVARY GLAND REGENERATION IN SJOGREN'S SYNDROME - PROJECT SUMMARY SJGREN’S SYNDROME IS AN AUTOIMMUNE DISEASE THAT CAUSES CHRONIC INFLAMMATION AND DAMAGE/DYSFUNCTION OF SALIVARY AND LACRIMAL GLANDS, WITH HYPOSALIVATION AS ONE OF THE MAIN CLINICAL MANIFESTATIONS. THE DISEASE GREATLY COMPROMISES THE ORAL AND SYSTEMIC HEALTH OF 4 MILLION AMERICANS, MOSTLY WOMEN, WITH NO CURE OR EFFECTIVE BIOLOGICAL THERAPY. STEM CELL THERAPIES HOLD GREAT PROMISE FOR SJGREN’S DISEASE, BUT THE CHRONIC AUTOIMMUNE INFLAMMATION GREATLY IMPEDES THE LIKELIHOOD OF SUCCESS. THERE IS A CRITICAL NEED FOR NEW STRATEGIES AND METHODS FOR ATTENUATION OF AUTOIMMUNE INFLAMMATION ACHIEVE EFFECTIVE AND SUSTAINED REGENERATION OF THE DAMAGED AND DYSFUNCTIONAL SALIVARY GLANDS. THE OBJECTIVE OF THIS EXPLORATORY PROJECT TO ELUCIDATE THE IMPACT OF SPECIALIZED PRO- RESOLVING MEDIATORS (SPMS) ON THE EXPANSION, RENEWAL AND IMMUNE-REGULATORY ACTIVITY OF SALIVARY GLAND STEM CELLS BOTH IN VITRO AND IN VIVO. SPMS ARE PRO-RESOLVING LIPID MEDIATORS THAT MEDIATE ACTIVE RESOLUTION OF INFLAMMATION AND HAVE DEMONSTRATED ABILITIES TO ENHANCE THE IMMUNOMODULATORY, PRO-HEALING AND REGENERATIVE PROPERTIES OF STEM CELLS. MOREOVER, SPMS CAN ALSO BE PRODUCED BY STEM CELLS. OUR PRELIMINARY STUDIES PROVIDED COMPELLING EVIDENCE THAT SPMS POSITIVELY IMPACT BOTH THE STEM CELL ACTIVITY AND THE IMMUNE-REGULATORY PROPERTY OF MOUSE SALIVARY GLAND STEM CELLS. OUR INNOVATIVE CENTRAL HYPOTHESIS IS THAT SPMS CAN PROMOTE SUSTAINABLE STRUCTURAL AND FUNCTIONAL RESTORATION OF DAMAGED SALIVARY GLANDS IN SJGREN’S DISEASE SETTING BY ACTING ON BOTH SALIVARY GLAND STEM CELLS AND IMMUNE/INFLAMMATORY CELLS TO SIMULTANEOUSLY MITIGATE AUTOIMMUNE INFLAMMATION AND ENHANCE TISSUE REGENERATION. IN AIM 1, WE WILL EXAMINE WHETHER LIPOXIN A4 AND MARESIN-1 CAN BOOST THE RENEWAL, EXPANSION, SALIVARY ORGANOID FORMATION, SPM PRODUCTION AND IMMUNE-REGULATORY ACTIVITY OF SALIVARY GLAND STEM CELLS FROM NORMAL C57BL/6 MICE IN MATRIGEL-BASED EXPANSION AND DIFFERENTIATION CULTURE SYSTEMS. IN AIM 2, WE WILL DETERMINE IF THESE SPMS CAN PROMOTE THE STRUCTURAL AND FUNCTIONAL RESTORATION OF DAMAGED SALIVARY GLANDS IN SJGREN’S DISEASE CONDITION IN VIVO. SPECIFICALLY, WE WILL ASSESS WHETHER LIPOXIN A4 AND MARESIN-1 ADMINISTERED TO SJGREN’S DISEASE-AFFLICTED MICE TRANSPLANTED WITH SALIVARY GLAND STEM CELLS CAN SIMULTANEOUSLY ATTENUATE INFLAMMATION AND ENHANCE TISSUE REGENERATION, THEREBY ACHIEVING SUSTAINABLE STRUCTURAL AND FUNCTIONAL RESTORATION OF SALIVARY GLANDS. MAJOR METHODOLOGIES EMPLOYED IN THIS PROJECT INCLUDE MATRIGEL CULTURE, INTRA- SALIVARY GLAND CELL TRANSFER, HIGH-THROUGHPUT RNA-SEQUENCING AND BIOINFORMATICS, FLOW CYTOMETRY, REAL-TIME QPCR, IMMUNOHISTOCHEMISTRY AND LUMINEX ASSAY. COMPLETION OF THIS INNOVATIVE EXPLORATORY PROJECT WILL ELUCIDATE THE ACTIONS OF SPMS IN SALIVARY GLAND STEM CELL-MEDIATED TISSUE REGENERATION IN THE SJÖGREN’S DISEASE CONDITION. IT WILL ADVANCE THE DEVELOPMENT OF STEM CELL THERAPIES THAT CONCOMITANTLY ATTENUATE AUTOIMMUNE INFLAMMATION AND ENHANCE TISSUE REGENERATION TO ACHIEVE SUSTAINABLE AND EFFECTIVE RESTORATION OF SALIVARY GLANDS FOR SJÖGREN’S SYNDROME PATIENTS AS WELL AS PATIENTS WITH OTHER SALIVARY GLAND INJURY/ INFLAMMATORY CONDITIONS.

A BIOMIMETIC STRATEGY TO TREAT ENAMEL LOSS

REGULATION OF SALIVARY GLAND INFLAMMATION IN SJOGREN'S SYNDROME BY ANNEXIN 1

SALIVA-MEDIATED MECHANISMS OF POST-ERUPTIVE ENAMEL MINERALIZATION

IDENTIFICATION AND CHARACTERIZATION OF ENDOGENOUS DENTAL PULP STEM CELLS - TITLE IDENTIFICATION AND CHARACTERIZATION OF ENDOGENOUS DENTAL PULP STEM CELLS ABSTRACT THE GOAL OF THIS PROPOSAL IS TO IDENTIFY AND CHARACTERIZE ENDOGENOUS DENTAL PULP STEM CELLS (DPSCS) IN ADULT MICE MOLARS. ALTHOUGH CONTINUOUSLY GROWING MOUSE INCISORS HAVE BEEN USED AS A MODEL SYSTEM TO CHARACTERIZE DENTAL STEM CELLS, ENDOGENOUS DPSCS WITHIN THE MOLARS REMAIN UNDERSTUDIED. UNLIKE INCISORS, MOUSE MOLARS ARE NOT CONTINUOUSLY GROWING AND RESEMBLE HUMAN TEETH. THEREFORE, IT IS CRUCIAL FOR THE FIELD OF STEM CELL BIOLOGY AS WELL AS DENTAL PULP REGENERATION THERAPY TO IDENTIFY AND CHARACTERIZE ENDOGENOUS DPSCS ISOLATED FROM THE MOLARS. TO CONDUCT THIS RESEARCH WE HAVE DEVELOPED A MOUSE MODEL IN WHICH AXIN2+ DENTAL PULP CELLS IN MOLARS ARE GENETICALLY LABELED. OUR PRELIMINARY ASSESSMENT SHOWS THAT AXIN2+ DPSCS ARE CAPABLE OF SELF-RENEWING AND DIFFERENTIATING INTO MATURE ODONTOBLASTS FOR A YEAR IN VIVO. IN ADDITION, THE TRANSPLANTED DENTAL PULP CELLS ARE CAPABLE REGENERATE PULP-LIKE TISSUE. THIS INITIAL DISCOVERY SHOWS A DIFFERENCE IN THE TYPE OF TISSUES REGENERATED FROM OTHER ADULT STEM CELLS AND DPSCS, SUGGESTING THEIR UNIQUE CELLULAR PLASTICITY PRESENT IN ENDOGENOUS DPSCS. IN THIS APPLICATION, WE WILL TEST IF AXIN2-EXPRESSING DPSCS CAN MEET THE CRITERIA OF ADULT STEM CELLS. WE WILL ALSO EXAMINE THE STEM CELLS PROPERTIES AT A SINGLE CELL LEVEL. THE REGENERATIVE ABILITY AND CELLULAR PLASTICITY OF THE TRANSPLANTED ENDOGENOUS DPSCS WILL BE CAREFULLY ASSESSED. POTENTIAL HETEROGENEITY OF AXIN2+ DPSCS WILL BE IDENTIFIED THROUGH SINGLE-CELL RNA-SEQUENCE ANALYSIS AND BIOINFORMATICS ANALYSIS. THE COMPLETION OF THIS PROPOSAL HAS OUTSTANDING POTENTIAL TO ADVANCE OUR KNOWLEDGE BASE OF ENDOGENOUS DPSCS IN MOLARS, LEADING TO NOVEL STRATEGIES FOR TISSUE REGENERATION THERAPIES.

REGULATION OF B-CELL APOPTOSIS BY TLR SIGNALING IN PERIODONTAL DISEASE

HIF-1ALPHA MEDIATED WOUND HEALING IN PERIAPICAL LESION

ROLE OF AMELOGENIN PHOSPHORYLATION IN REGULATING ENAMEL FORMATION IN VIVO

PROOF OF CONCEPT: SCARDOVIA WIGGSIAE IS CARIOGENIC IN VIVO

MECHANISMS AND THERAPEUTIC MODULATION OF T CELL AUTOIMMUNE RESPONSES IN SJOGREN'S SYNDROME

NANOPARTICLE-BASED ANTIMICROBIAL PHOTOCHEMOTHERAPY IN BIOFILMS

MECHANISMS OF PEROXISOME PROLIFERATOR-ACTIVATED RECEPTOR-ALPHA REGULATION IN PERIDONTITIS - PROJECT SUMMARY THIS APPLICATION IS FOR THE EXTENSION OF OUR CURRENT R03 GRANT TITLED “PATHOGENIC ROLE OF PEROXISOME PROLIFERATOR- ACTIVATED RECEPTOR ALPHA IN PERIODONTITIS”, WHICH HAS PRODUCED 2 PUBLISHED AND 1 UNDER REVIEW PUBLICATIONS SO FAR. OUR STUDIES HAVE FOCUSED ON THE PATHOGENIC ROLE OF PPARA IN PERIODONTAL INFLAMMATION AND BONE RESORPTION. SPECIFICALLY, WE DEMONSTRATED THAT PPARA NOT PPARSS OR PPAR HAD DECREASED EXPRESSION LEVELS IN GINGIVAL CELLS STIMULATED WITH PERIODONTITIS ANTIGENS AND IN GINGIVAL TISSUE OF MOUSE EXPERIMENTAL PERIODONTITIS, AND PPARA AGONIST TREATMENT SIGNIFICANTLY REDUCES INFLAMMATION AND BONE LOSS IN EXPERIMENTAL PERIODONTITIS. WHILE THE MECHANISM OF HOW ACTIVATED PPARA REGULATES THESE PATHOLOGY OF PERIODONTITIS IS LARGELY UNKNOWN. OUR RECENT DATA DEMONSTRATED THAT ACTIVATION OF PPARA INHIBITED THE NF-KB (P65) SIGNALING AND INCREASED THE ANTI- INFLAMMATORY CYTOKINES IL-10 AND ITS SECRETION FACTOR CD36 IN THE MACROPHAGES. THE WNT PATHWAY WAS ABERRANTLY ACTIVATED IN THE MACROPHAGE DURING PERIODONTITIS IN VIVO AND ACTIVATED PPARA DECREASED WNT/SS-CATININ/TCF-4 ACTIVITY AND INDUCED A BINDING OF SS-CATENIN WITH PPARA IN IN VITRO STUDIES. TOGETHER WITH THE PREVIOUSLY PUBLISHED FINDINGS BY OTHERS, WE HYPOTHESIZE THAT PPARA IS AN EFFECTIVE MACROPHAGE MODULATOR THROUGH DIVERSION THE WNT/SS-CATENIN SIGNALING VIA COMPETING WITH TCF4 FOR BINDING TO SS-CATENIN, ENHANCING THE CD36/IL-10 MEDIATED ANTI-INFLAMMATORY TRANSCRIPTIONAL ACTIVITIES AND SUPPRESSING THE TCF4 MEDIATED PRO- INFLAMMATORY TRANSCRIPTIONAL ACTIVITIES. IN THIS PROPOSAL, WE WILL FIRST DETERMINE IF PPARA AGONIST’S PERIODONTITIS PROTECTIVE EFFECT IS VIA MODULATING MACROPHAGE POLARIZATION THROUGH A PPARA AGONIST DEPENDENT MECHANISM (AIM 1); THEN WE WILL EXPLORE THE UNDERLYING MECHANISM BY WHICH PPARA MODULATES MACROPHAGE IN THE PERIODONTITIS (AIM 2). SUCCESSFUL COMPLETION OF THIS PROJECT WILL GENERATE CONCEPTUAL ADVANCES IN OUR UNDERSTANDING OF THE ETIOLOGY OF THE DISEASE BY HIGHLIGHTING PREVIOUSLY POORLY UNDERSTOOD PATHWAYS OF INFLAMMATION. IF OUR HYPOTHESIS IS CORRECT, THESE STUDIES WILL BROADEN OUR INSIGHTS INTO THE POTENTIAL ROLE OF PPARA IN THE REGULATION OF MACROPHAGES IN THE INFLAMMATION OF PERIODONTAL DISEASE, AS WELL AS OTHER IMMUNE-MEDIATED OSTEOLYTIC CONDITIONS SUCH AS OSTEOPOROSIS OR RHEUMATOID ARTHRITIS.

ENAMELYSIN PROCESSING MECHANISMS IN AMELOGENESIS

OSTEOBLAST DOPAMINE RECEPTOR MEDIATES DIABETIC BONE DISEASE - ABSTRACT DIABETES IS ESTIMATED TO AFFECT AT LEAST 100 MILLION PEOPLE IN THE UNITED STATES. DIABETIC BONE DISEASE IS A COMPLICATION OF BOTH TYPE 1 AND TYPE 2 DIABETES. DIABETIC BONE DISEASE RESULTS IN DECREASED MOBILITY DUE TO INCREASED FRACTURE RISK WITH FURTHER COMPLICATIONS THAT CAN ULTIMATELY BE FATAL. LOW BONE MINERAL DENSITY THAT IS OFTEN USED TO DIAGNOSE DIABETIC BONE DISEASE DOES NOT CORRELATE WELL WITH THE ACTUAL IMPAIRMENT IN MECHANICAL PROPERTIES OBSERVED IN TYPE 1 AND TYPE 2 DIABETES. BY CONTRAST, COLLAGEN IN DIABETIC BONE IS WELL UNDERSTOOD TO HAVE LOW LEVELS OF LYSINE-DERIVED BIOSYNTHETIC CROSS-LINKS WHICH ARE ESSENTIAL FOR THE PHYSICAL STRENGTH OF CONNECTIVE TISSUES, INCLUDING BONE. HOWEVER, FEW STUDIES OF LEVELS AND REGULATION OF LYSYL OXIDASE IN DIABETIC BONE EXIST. RECENT INTEREST IN GUT-DERIVED ENDOCRINE FACTORS HAS DEVELOPED FROM CONSEQUENCES OF BARIATRIC SURGERY TO TREAT OBESE INDIVIDUALS, MANY OF WHOM ALSO SUFFER FROM DIABETES. GLUCOSE INSULINOTROPIC POLYPEPTIDE (GIP) IS A GUT-DERIVED HORMONE KNOWN AS AN INCRETIN. IN ADDITION TO TARGETING PANCREATIC SS CELLS, GIP DIRECTLY TARGET OSTEOBLASTS VIA THE GIP RECEPTOR (GIPR), STIMULATING A BONE ANABOLIC RESPONSE. THE GUT ALSO SECRETES DOPAMINE INTO THE PERIPHERAL CIRCULATION THAT HAS BEEN HYPOTHESIZED TO INHIBIT THE ACTIVITY OF GIP, MEDIATED BY DOPAMINE RECEPTORS, ALSO PRESENT ON OSTEOBLASTS. HERE WE INTEGRATE THESE FINDINGS AND OUR OWN DATA INTO THE FOLLOWING PARADIGM-SHIFTING PROPOSAL. OUR HYPOTHESIS IS THAT NORMAL ANABOLIC STIMULATION OF BONE FORMATION AND LYSYL OXIDASE PRODUCTION BY GIP IS DYSREGULATED IN DIABETES BY GUT-DERIVED DOPAMINE MEDIATED BY THE OSTEOBLAST DOPAMINE D2 RECEPTOR (D2R) IN BOTH TYPE 1 AND TYPE 2 DIABETES. WE PROPOSE TO TEST THIS HYPOTHESIS IN MALE AND FEMALE C57BL6/J CONDITIONAL OSTEOBLAST-SPECIFIC D2R GENE KNOCKOUT MICE (COL1-2.3-CRE CROSS-BRED WITH D2RFL/FL MICE). MICE WILL BE SUBJECTED TO DEVELOPMENT OF BOTH TYPE 1 AND TYPE 2 DIABETES BY, RESPECTIVELY, MULTIPLE LOW DOSE STREPTOZOTOCIN INJECTIONS (TYPE 1 DIABETES), AND TO HIGH FAT DIET (TYPE 2 DIABETES). ANALYSES WILL INCLUDE MEASURES OF BONE MARKERS BY QPCR PLUS ALL FIVE LYSYL OXIDASE FAMILY MRNAS, BONE STRUCTURE BY ΜCT, BONE STRENGTH, BONE HISTOLOGY FOR GROWTH PLATE STRUCTURE, AND A VARIETY OF RELEVANT SERUM MARKERS. OUR HYPOTHESIS PREDICTS THAT OSTEOBLAST D2R GENE KNOCKOUT MICE WILL BE RESISTANT TO BOTH TYPE 1 AND TYPE 2 DIABETES-INDUCED BONE PATHOLOGY, COMPARED TO CONTROLS. THE HIGH POTENTIAL IMPACT OF THE PROPOSAL IS TO DEFINITIVELY IDENTIFY A CRITICAL ASPECT OF AN UNEXPECTED METABOLIC RELATIONSHIP THAT DRIVES A SIGNIFICANT BONE COMPLICATION OF BOTH TYPE 1 AND TYPE 2 DIABETES THAT WE CONTEND WILL HAVE TRANSLATIONAL IMPLICATIONS FOR AN INCREASINGLY IMPORTANT PUBLIC HEALTH PROBLEM. DELIVERABLES: A PREVIOUSLY UNEXPLORED METABOLIC RELATIONSHIP BETWEEN THE GUT AND BONE THAT IS DYSREGULATED IN DIABETES WILL THUS BE DEFINITIVELY IDENTIFIED AND SUPPORTED BY SUBJECTING A SINGLE GENETIC MOUSE MODEL TO TYPE 2 AND TYPE 1 DIABETES. DATA WILL POINT TO NOVEL DRUGGABLE MOLECULAR TARGETS. NOVEL MECHANISTIC INSIGHTS INTO DIABETIC BONE DISEASE WILL BE GAINED.

HYPOXIA AND PMN FUNCTION IN PERIODONTITIS

ROLE OF NHA-OC/NHA2 IN OSTEOCLAST DIFFERENTIATION AND FUNCTION

REGULATORY B CELLS IN THE AMELIORATION OF IMMUNE-MEDIATED PERIODONTAL DISEASE

HEALTH CARE AND OTHER FACILITIES

PH-SENSITIVE MATERIALS RESPONDING TO METABOLIC ACTIVITIES OF CARIOGENIC PLAQUE

HEALTH CARE AND OTHER FACILITIES

FLOW CYTOMETER / CELL SORTER

THE ESSENTIAL ROLE OF MIR-27A IN CRANIOFACIAL AND BODY SKELETONS.

HOSPITAL VOLUME FOR OROFACIAL CLEFT REPAIR AND RISK OF COMPLICATIONS

GUT MICROBIOME AND SALIVARY GLAND FUNCTION: PROTECTIVE ACTIONS & KEY PLAYERS - PROJECT SUMMARY SALIVARY GLAND FUNCTION IS CHRONICALLY IMPAIRED IN SJӦGREN’S SYNDROME, AN AUTOIMMUNE EXOCRINE GLAND DISEASE WITH NO CURE OR EFFECTIVE TREATMENT AVAILABLE. ABERRANTLY ACTIVATED INNATE IMMUNE CELLS, AUTOREACTIVE T- AND B CELLS, AND PROINFLAMMATORY CYTOKINES COLLECTIVELY CAUSE SJOGREN’S-CHARACTERISTIC SALIVARY GLAND INFLAMMATION AND DYSFUNCTION. THERE IS AN URGENT NEED FOR NOVEL, EFFECTIVE THERAPIES THAT PROTECT/IMPROVE SALIVARY GLAND FUNCTION AND AMELIORATE SALIVARY GLAND INFLAMMATION. THE IMPORTANCE OF HOST GUT MICROBIOME IN HEALTH AND DISEASE IS INCREASINGLY RECOGNIZED, YET VERY LITTLE IS KNOWN ABOUT THE IMPACT OF GUT MICROBIOME ON SALIVARY GLANDS, ESPECIALLY THE PROTECTIVE BACTERIA AND THEIR ACTIONS. HENCE, THE MAIN OBJECTIVE OF THIS PROJECT IS TO ADDRESS THIS KNOWLEDGE GAP, WITH A PARTICULAR FOCUS ON IDENTIFYING THE POTENTIALLY SALIVARY GLAND-PROTECTIVE GUT BACTERIA AND REVEALING THEIR PRECISE ACTIONS USING BOTH MURINE MODELS AND CELLS AND SAMPLES FROM SJӦGREN’S DISEASE PATIENTS. OUR PRELIMINARY STUDIES SHOWED THAT TRANSPLANT OF FECAL BACTERIA FROM HEALTHY C57BL/6 MICE TO THE NON-OBESE DIABETIC MICE, A SPONTANEOUS MODEL OF SJӦGREN’S SYNDROME, EXERTED A PROTECTIVE EFFECT ON SALIVARY GLANDS. THE PROTECTION WAS ACCOMPANIED BY REDUCED INFLAMMATORY RESPONSES AND DIMINISHED EXPRESSION OF BROMODOMAIN CONTAINING 2, AN EPIGENETIC READER/MODULATOR, IN SALIVARY GLAND EPITHELIAL CELLS. WE ALSO IDENTIFIED MULTIPLE SPECIFIC BACTERIAL SPECIES WITH SALIVARY GLAND-PROTECTIVE POTENTIALS BASED ON OUR PRELIMINARY MICROBIOME ANALYSES AND THE PERTINENT EVIDENCE FROM HUMAN STUDIES IN THE LITERATURE. WE FORMED THE CENTRAL HYPOTHESIS THAT HEALTHY GUT MICROBIOTA CONTAINS BACTERIAL SPECIES THAT, ACTING THROUGH THEIR METABOLITES, PROTECT SALIVARY GLAND FUNCTION BY MITIGATING INFLAMMATORY RESPONSES AND DYSFUNCTION OF SALIVARY GLAND EPITHELIAL CELLS THROUGH MECHANISMS INCLUDING DOWNREGULATING THE EXPRESSION OF EPIGENETIC READER OF BROMODOMAIN CONTAINING 2. AIM 1 OF THIS PROJECT WILL DEFINE AND DISCOVER THE MECHANISMS UNDERLYING THE PROTECTIVE EFFECTS OF HEALTHY C57BL/6 FECAL MICROBIOTA TRANSPLANT ON SALIVARY GLANDS IN THE NON-OBESE DIABETIC MICE AND A SECOND, INDUCED MODEL OF SJӦGREN’S SYNDROME. AIM 2 WILL EXPLORE THE PROTECTIVE IMPACT OF SPECIFIC GUT BACTERIAL SPECIES ON SALIVARY GLAND FUNCTION IN THE TWO MOUSE MODELS. AIM 3 WILL IDENTIFY AND TEST MICROBIAL METABOLITES THAT MAY MEDIATE THE SALIVARY GLAND- PROTECTIVE IMPACT OF GUT MICROBIOTA USING BOTH MOUSE SS MODELS AND CELLS/SAMPLES FROM HUMAN SJӦGREN’S DISEASE PATIENTS. THE PROJECT WILL EMPLOY A COMBINATION OF SOPHISTICATED IMMUNOLOGICAL, MICROBIOLOGICAL AND SYSTEMIC BIOLOGICAL METHODOLOGIES, INCLUDING 16S RRNA GENE SEQUENCING ANALYSIS OF MICROBIOME, RNA- SEQUENCING ANALYSIS OF HOST CELLS, AND METABOLOMIC ASSAYS. KNOWLEDGE GENERATED WILL HAVE DIRECT RELEVANCE TO AND IMPACT ON HUMAN HEALTH, AND WILL ADVANCE THE DEVELOPMENT OF NOVEL, GUT MICROBE/MICROBIAL METABOLITE- BASED THERAPIES TO COMBAT AUTOIMMUNE SJӦGREN’S DISEASE AS WELL AS OTHER SALIVARY GLAND INFLAMMATORY DISORDERS.

MECHANISTIC INVESTIGATION OF MULTISPECIES INTERACTIONS IN CLEAR ALIGNER INDUCED PERIODONTAL INFLAMMATION - ABSTRACT CLEAR ALIGNER TREATMENT CAN LEAD TO ORAL MICROBIAL SHIFT WHICH MAY IMPACT PERIODONTAL HEALTH. THE INTERPLAY BETWEEN MICROBES AND THE HOST PLAYS CRITICAL ROLES IN HEALTH AND DISEASES. HOWEVER, A MECHANISTIC UNDERSTANDING OF HOW THE SHIFTED MICROBIOME (DYSBIOSIS) AND ITS INTERACTION WITH THE HOST INFLUENCE ORAL HEALTH IS STILL LACKING. OUR PRELIMINARY STUDIES DEMONSTRATED THE INCREASED RELATIVE ABUNDANCE OF FUSOBACTERIUM NUCLEATUM (FN), ACTINOMYCES SPP. AND SACCHARIBACTERIA (TM7) POSITIVELY CORRELATED WITH THE PROGRESSION OF GINGIVITIS IN CLEAR ALIGNER PATIENTS. SPECTRAL FISH IMAGINING OF CLINICAL SAMPLES AND IN VITRO CO-AGGREGATION ANALYSIS INDICATED A CLOSE PHYSICAL ASSOCIATION BETWEEN FN, ACTINOMYCES SPP., AND TM7. MORE IMPORTANTLY, WE PROVIDED NEW PRELIMINARY DATA TO DEMONSTRATE THE POTENTIAL SYNERGY AMONG THE THREE BACTERIAL SPECIES VIA PHYSICAL INTERACTION AND METABOLIC CROSS-FEEDING. THE PROPOSED STUDY FOCUSES ON THESE THREE SPECIES WHICH HAVE A MYRIAD OF VIRULENCE FACTORS RELEVANT TO ORAL AND SYSTEMIC DISEASES AND POSITIVELY ASSOCIATED WITH PERIODONTAL INFLAMMATORY DISEASES. THESE INTRIGUING FINDINGS SERVE AS THE SCIENTIFIC PREMISE FOR THE WORKING HYPOTHESIS THAT FN, ACTINOMYCES SPP. AND TM7 MAY ENGAGE IN SYNERGISTIC INTERACTIONS TO ENHANCE THEIR GROWTH WITHIN BIOFILM AND EXHIBIT COOPERATIVE EFFECTS WHEN INTERACTING WITH HOST ORAL EPITHELIAL CELLS TO PROMOTE INFLAMMATION. THE WELL- STUDIED TYPE STRAINS FN ATCC 23726, A. ODONTOLYTICUS XH001 AND NANOSYNBACTER LYTICUS STRAIN TM7X, WHICH REPRESENT FN, ACTINOMYCES SPP., AND TM7, RESPECTIVELY, WILL BE USED IN THIS STUDY. TWO INDEPENDENT BUT INTERCONNECTED AIMS ARE DEVELOPED TO TEST THE WORKING HYPOTHESIS: AIM 1. INVESTIGATE THE SYNERGISM BETWEEN FN, A. ODONTOLYTICUS, AND TM7 DURING BIOFILM FORMATION. USING FN 23726, XH001 AND TM7X MODEL STRAINS, COMPREHENSIVE IMAGING-BASED ANALYSIS WILL BE EMPLOYED TO INVESTIGATE THE SYNERGISTIC INTERACTION DURING BIOFILM GROWTH BY ASSESSING BIOFILM BIOVOLUME, MAXIMAL THICKNESS, ROUGHNESS CORRELATION RATIO, AND CONTINUITY RATIO VIA CONFOCAL LASER SCANNING MICROSCOPY (CLSM). AIM 2. EXAMINE THE COOPERATIVE EFFECTS BETWEEN FN, A. ODONTOLYTICUS, AND TM7 WHEN INTERACTING WITH HOST ORAL EPITHELIAL CELLS. WE WILL USE AN IN VITRO CULTURE SYSTEM WITH VARIOUS COMBINATIONS OF FN 23726, XH001 AND TM7X WILDTYPE STRAINS IN THE PRESENCE OR ABSENCE OF HUMAN ORAL KERATINOCYTES (HOK-16B) TO IDENTIFY GENES WITH EXPRESSION CHANGES SPECIFICALLY ATTRIBUTABLE TO THE SYNERGY OF THREE SPECIES DURING INTERACTIONS WITH HOK- 16B CELLS AND DETERMINE IF TRI-SPECIES INTERACTION HAS COOPERATIVE EFFECTS CONTRIBUTING TO GINGIVAL INFLAMMATION. THE SUCCESSFUL COMPLETION OF THESE AIMS WILL PROVIDE NOVEL MECHANISTIC-LEVEL INSIGHT INTO THE IMPACT OF MICROBIAL SHIFT DURING TREATMENT WITH CLEAR ALIGNERS ON PERIODONTAL HEALTH. MORE IMPORTANTLY, THE STUDY WILL HELP IDENTIFY THE MOLECULAR TARGETS INVOLVED IN POLYMICROBIAL SYNERGY CONTRIBUTING TO PERIODONTAL DISEASE INITIATION/PROGRESSION FOR POTENTIAL THERAPEUTIC PURPOSES IN THE FUTURE.

PATHOGENIC ROLE OF PEROXISOME PROLIFERATOR-ACTIVATED RECEPTOR ALPHA IN PERIODONTITIS - PROJECT SUMMARY PERIODONTITIS IS AN IMPORTANT PUBLIC HEALTH PROBLEM AMONG ADULTS IN THE U.S., WITH MAJOR ECONOMIC COSTS FOR PREVENTION AND TREATMENT AND SIGNIFICANT IMPACT ON QUALITY OF LIFE. UN-CONTROLLED PERIODONTITIS CAN CAUSE EX- TENSIVE TOOTH LOSS, JAW BONE DETERIORATION, AND INCREASED RISK OF DEVELOPING SYSTEMIC DISEASES. CONVENTIONAL TREATMENTS THAT RELY ON ANTIBIOTICS AND MECHANICAL REMOVAL OF DENTAL PLAQUE ARE TRANSIENTLY EFFECTIVE BECAUSE THEY INDIRECTLY ADDRESS THE INFLAMMATION AND RELATED IMMUNE RESPONSES THAT UNDERLIE PERIODONTITIS, BUT DO NOT DIRECTLY IMPACT PATHOGENESIS. THUS, THERE IS A COMPELLING NEED TO INVESTIGATE NOVEL TARGET MOLECULES WHICH DI- RECTLY MODULATE PATHOGENESIS FOR BOTH INFLAMMATION AND BONE LOSS IN PERIODONTITIS. IT HAS BEEN DEMONSTRATED THAT PPARA AGONISTS (PPARAA), WHICH IS PPARA SPECIFIC, HAVE ROBUST PROTECTIVE ACTIONS LIMITING INFLAMMATION IN AUTOIMMUNE DISEASE, MODULATING INFLAMMATION. OUR PRELIMINARY DATA SUGGEST THAT A PPARAA, FENOFIBRATE, HAS THE POTENTIAL TO REDUCE PERIODONTAL INFLAMMATION AND BONE RESORPTION IN EXPERIMENTAL PERIODONTITIS MOUSE MOD- ELS, SUGGESTING THAT THE PATHOLOGICAL ROLE OF PPARA IN PERIODONTITIS DESERVES FURTHER INVESTIGATION. WE PROPOSE TO TEST THE HYPOTHESIS THAT PPARA NOT PPARSS OR PPAR PLAYS IMPORTANT PATHOLOGICAL ROLES IN PERIODONTITIS. TO THAT END, TWO DISTINCT BUT COMPLEMENTARY SPECIFIC AIMS ARE PROPOSED. AIM1 WILL DETERMINE THAT PPARA NOT PPARSS OR PPAR HAS EXPRESSION LEVEL CHANGES IN PERIODONTITIS. AIM 2 WILL DETERMINE PPARA KNOCKOUT OR OVEREXPRES- SION WILL AFFECT INFLAMMATION AND BONE LOSS IN EXPERIMENTAL PERIODONTITIS MOUSE MODEL. THE UNDERSTANDING OF EXPRESSION LEVEL CHANGE OF PPARA IN PERIODONTITIS INVESTIGATED DURING AIM1 WILL PROVIDE THE FOUNDATION FOR AIM2. THE GOAL IS TO REVEAL THE ROLE OF PPARA IN INFLAMMATORY REGULATION AND IN MODULATION OF BONE HOMEOSTASIS DURING PERIODONTAL DISEASES. BUILDING ON PREVIOUS EXPERIENCE AND BOTH AIM1 AND AIM2 WILL LEAD TO THE DEVELOPMENT OF NOVEL, PPARA TARGETED THERAPIES FOR PERIODONTITS AND VARIOUS OTHER ORAL DISEASES. SUCCESSFUL COMPLETION OF THIS PROJECT WILL LEAD TO BETTER UNDERSTANDING OF THE MOLECULAR AND CELLULAR ROLE OF PPARA IN PERIODONTITIS, AND TRANSLATE THIS KNOWLEDGE TO DEVELOP AN APPLICATION SYSTEM TO ACHIEVE SUSTAINED RELEASE WITH NONINVASIVE LOCAL DELIVERY FOR THE CLINICAL TREATMENT OF PERIODONTITIS.

WHAT GIVES THE DENTIN-ENAMEL JUNCTION STRENGTH? STRUCTURAL AND MECHANICAL FUNCTION OF COLLAGEN AND AMELOGENIN.

THE ROLE OF PLASMACYTOID DENTDRITIC CELLS IN THE PATHOGENESIS OF SJOGREN'S SYNDROME

FROM SYMBIOSIS TO DYSBIOSIS AND BACK: THE IMPACT OF PRO-INFLAMMATION AND PRO-RESOLUTION - PERIODONTITIS IS INITIATED BY ORAL BACTERIA, BUT QUICKLY TRANSITS TO AN INFLAMMATORY IMMUNE DISEASE WITH LOSS OF HOST- BACTERIAL HOMEOSTASIS. HOMEOSTASIS, THE SYMBIOTIC BALANCE BETWEEN HOST TISSUES AND THE RESIDENT PERIODONTAL MICROBIOME IS DISRUPTED BY A DYSREGULATED IMMUNE RESPONSE. FROM SYMBIOSIS TO DYSBIOSIS, EARLY INNATE IMMUNE COMPONENTS ARE ROBUSTLY LINKED. AT THE HOST-MICROBIAL INTERFACE, WE HAVE AMPLE EVIDENCE THAT MICROBES TRIGGER THE IMMUNE RESPONSE BUT LACK KNOWLEDGE OF HOW THE IMMUNE RESPONSE REGULATES THE COMPOSITION AND FUNCTIONAL ACTIVITY OF THE MICROBIOME. SPECIALIZED PRO-RESOLVING MEDIATORS (SPMS) ARE A FAMILY OF EICOSANOID LIPID MEDIATORS THAT BIND TO SPECIFIC CELL RECEPTORS TO COORDINATE THE RETURN OF AN INFLAMMATORY LESION TO HOMEOSTASIS. AMONG THE MANY SPMS, E-SERIES RESOLVINS ARE NATURALLY OCCURRING METABOLITES OF TRANSFORMATION CIRCUITS OF THE OMEGA-3 FATTY ACID, EICOSAPENTAENOIC ACID (EPA), THAT ACTIVELY INDUCE RESOLUTION OF INFLAMMATION. ACTIVATION OF THE RESOLVIN E1 (RVE1) RECEPTOR (ERV1) IN GAIN-OF-FUNCTION STUDIES WITH THE ERV1 TRANSGENIC MOUSE (ERV1TG), SHOW PROFOUND RESISTANCE TO PERIODONTITIS. THE USE OF RESOLVINS TO CONTROL INFLAMMATION ALLOWS US TO TEST OUR CENTRAL HYPOTHESIS THAT EARLY IMMUNE ACTIVATION DIRECTLY CONTRIBUTES TO THE DEVELOPMENT OF PERIODONTAL DYSBIOSIS, WHICH CAN BE RE- VERSED/PREVENTED BY ACTIVATING RESOLIVIN-E1 MEDIATED RESOLUTION OF INFLAMMATION PATHWAYS. TO TEST THIS HYPOTH- ESIS, WE BREAK OUR INVESTIGATIONS INTO TWO LEVELS. FIRST, IN A MURINE MODEL, WE WILL CHARACTERIZE THE ORAL MICROBI- OME’S FUNCTIONAL AND SPATIAL ORGANIZATIONAL CHANGES AS SYMBIOSIS TRANSITS TO DYSBIOSIS. SECOND, WE WILL USE AN IN VITRO SYSTEM TO UNRAVEL THE MECHANISMS THAT DETERMINE HOW RVE1 AND ITS RECEPTOR (ERV1) INFLUENCE THE DY- NAMICS OF DYSBIOSIS AND INFLAMMATORY/IMMUNE RESPONSES IN PERIODONTITIS. THE CANDIDATE IS A DDS/PHD PERIO- DONTIST WHO IS COMMITTED TO AN ACADEMIC CAREER DEDICATED TO IMPROVING PERIODONTAL AND ORAL HEALTH. THE MEN- TORED PHASE OF THE PROPOSAL WILL BE CARRIED OUT WITH DR. THOMAS VAN DYKE (PRIMARY MENTOR) AND DR. XUESONG HE (CO-MENTOR) AT THE FORSYTH INSTITUTE. THE GOAL OF THE K99 PHASE IS TO PROVIDE ADDITIONAL TRAINING, MENTORING, AND EXPERIENCE IN THREE IMPORTANT AREAS 1) ADVANCED BIOINFORMATICS TRAINING FOR MICROBIOME DATA; 2) STUDY DESIGN AND BIOSTATISTICS TRAINING; AND 3) CAREER DEVELOPMENT. A STRONG SCIENTIFIC ADVISORY COMMITTEE FROM THE FORSYTH INSTITUTE, HARVARD MEDICAL SCHOOL, AND THE UNIVERSITY OF WASHINGTON HAS BEEN ESTABLISHED WITH DIVERSE EXPERTISE IN IMMUNOLOGY, MICROBIOLOGY, BIOINFORMATICS, AND STATISTICS TO HELP GUIDE THE CANDIDATE PROVIDING A TRAINING PLAN THAT COMPLEMENTS AND EXPANDS HER SCIENTIFIC BACKGROUND WITH MENTORS WHOSE EXPERTISE SPANS THE BROAD SCOPE OF THE SCIENTIFIC PROBLEM. THE SCIENTIFIC COMPONENT AND TRAINING OUTLINED IN THIS PROPOSAL PROVIDE A PATHWAY FOR THE CANDIDATE TO TRANSIT TO AN INDEPENDENT RESEARCH CAREER IN A TENURE TRACK FACULTY POSITION INDEPENDENT OF THE RESEARCH PROGRAMS OF THE MENTORS. SUCCESSFUL COMPLETION OF THIS PROJECT WILL LEAD TO BETTER UNDERSTANDING OF HOST-MICROBIOME INTERACTIONS AND HOW TO GAIN CONTROL OF THE INFLAMMATION/DYSBIOSIS CYCLE IN PERIODONTITIS TO AME- LIORATE DISEASE. 1

DIFFERENTIATION OF MICROBE-MICROBE INTERACTIONS AT SINGLE-CELL LEVEL THROUGH UNCONVENTIONAL UTILIZATION OF EXPANSION MICROSCOPY - THE ORAL MICROBIOME PLAYS A CRUCIAL ROLE NOT ONLY IN ORAL HEALTH BUT ALSO IN SYSTEMIC HEALTH. DYSFUNCTIONAL ORAL MICROBIOMES HAVE BEEN LINKED TO VARIOUS CHRONIC HUMAN DISEASES. HOWEVER, UNDERSTANDING HOW THE ORGANIZATION OF THE ORAL MICROBIOME IS CONNECTED TO ITS FUNCTION REMAINS A MAJOR CHALLENGE. NEXT-GENERATION DNA SEQUENCING HAS REVOLUTIONIZED THE STUDY OF ORAL MICROBIAL COMMUNITIES BY PROVIDING COMPREHENSIVE INFORMATION ABOUT THEIR COMPOSITION. CLASI-FISH HAS CONTRIBUTED TO OUR UNDERSTANDING BY REVEALING THE SPATIAL ORGANIZATION OF THE MICROBIOME. WHILE IMAGING CAN INFORM US ABOUT WHICH MICROBES ARE IN PROXIMITY TO EACH OTHER AT THE MICRON SCALE, IT CANNOT ALONE DIFFERENTIATE WHETHER THIS SPATIAL PROXIMITY IS DUE TO FUNCTIONAL INTERACTIONS OR CHANCE OCCURRENCES. TO OVERCOME THIS LIMITATION, OUR PROPOSED STUDY AIMS TO INVESTIGATE MICROBE-MICROBE ADHESION AS A FUNCTIONAL ATTRIBUTE OF THEIR INTERACTIONS. WE WILL EMPLOY A COMBINATION OF EXPANSION MICROSCOPY (EXM) AND MULTIPLEX FISH IMAGING, WHICH WILL PROVIDE A UNIQUE OPPORTUNITY TO GLOBALLY ANALYZE MICROBIAL INTERACTIONS AT THE MICRON LEVEL. THIS APPROACH WILL ALLOW US TO EXPLORE THE FUNCTIONAL ASPECTS OF MICROBIAL COMMUNITIES BEYOND THEIR SPATIAL ARRANGEMENT. FURTHERMORE, THE PRESENCE OF THICK BIOFILMS OR AGGREGATES WITHIN THEIR ECOLOGICAL NICHES PRESENTS SIGNIFICANT CHALLENGES FOR SINGLE-CELL DISSECTION AND IMAGING, OFTEN LEADING TO DIFFICULTIES OR AMBIGUITIES IN TAXONOMIC ASSIGNMENT. OUR PROPOSED STUDY WILL ADDRESS THIS CONCERN BY QUANTIFYING THE RELATIVE MICROBIAL ADHESION FORCES AT THE SINGLE-CELL LEVEL. THIS WILL ENHANCE OUR UNDERSTANDING OF THE FORCES GOVERNING MICROBIAL INTERACTIONS WITHIN BIOFILMS AND CONTRIBUTE TO UNRAVELING THE COMPLEXITY OF THE ORAL MICROBIOME. OVERALL, THIS STUDY AIMS TO INVESTIGATE THE FUNCTIONAL ATTRIBUTES OF THE ORAL MICROBIOME AT THE MICRON SCALE, BRIDGING THE GAP BETWEEN MICROBIAL ORGANIZATION AND FUNCTION. BY EMPLOYING ADVANCED MICROSCOPY TECHNIQUES AND QUANTIFYING MICROBIAL ADHESION FORCES, WE HOPE TO GAIN VALUABLE INSIGHTS INTO THE INTRICATE DYNAMICS OF THE ORAL MICROBIOME AND ITS IMPLICATIONS FOR HUMAN HEALTH. THERE ARE THREE INTERCONNECTED AIMS: 1) TO COMBINE MULTIPLEX FISH IMAGING WITH EXPANSION MICROSCOPY. HERE, THE FOCUS WILL BE INVESTIGATING THE FEASIBILITY OF COMBINING MULTIPLEX PEPTIDE NUCLEIC ACID (PNA) FISH IMAGING WITH EXM ON AN IN VITRO POLYMICROBIAL BIOFILM. 2) TO QUANTIFY RELATIVE MICROBE-MICROBE ADHESION FORCE. HERE, EXPANSION FORCES WITHIN THE GEL MATRICES WILL BE CALIBRATED THROUGH FORCE-REFERENCE BEAD STRATEGY. FOLLOWING PNA FISH HYBRIDIZATION ON THE POLYMICROBIAL BIOFILM, THE MICROBIAL ADHESION FORCES WITHIN THE BIOFILM WILL BE QUANTIFIED BY COMPARISON WITH GEL EXPANSION FORCES. 3) TO EVALUATE SPECIES-LEVEL ADHESION FORCES WITHIN NATURAL HUMAN ORAL BIOFILMS. HERE, WE WILL VALIDATE OUR ADHESION FORCE MEASUREMENT APPROACH USING A DIVERSE RANGE OF IN VITRO COAGGREGATION PAIRS WITH ESTABLISHED BINDING PHENOTYPES. THEN SPECIES-LEVEL ADHESIVE FORCES WITHIN HUMAN NATURAL ORAL BIOFILMS WILL BE OBTAINED. THIS INDEPENDENCE AWARD (PA-20-188) WILL FOSTER MY INDEPENDENT RESEARCH GROWTH AND ALLOW ME TO ACCOMPLISH MY LONG-TERM CAREER GOAL OF BECOMING A PRODUCTIVE, INDEPENDENT TRANSLATIONAL SCIENTIST IN THE FIELD OF ORAL MICROBIOLOGY.

THE ROLE OF CD4+ MEMORY T CELL SUBTYPES IN PERIODONTAL DISEASE RECURRENCE - PROJECT SUMMARY THE RECURRENCE OF PERIODONTAL DISEASE (PD) IS A POORLY IMMUNOLOGICALLY DEFINED OR THERAPEUTICALLY TARGETED CLINICAL CHALLENGE. UNCOVERING THE HOMEOSTATIC AND PATHOGENIC POTENTIAL OF LONG-TERM IMMUNE CELLS IN PERIODONTAL TISSUES IS CRUCIAL TO REVEALING THE CELLULAR DRIVERS OF PD RECURRENCE. THIS PROPOSAL FOCUSES ON AN OVERLOOKED PERIODONTAL-ASSOCIATED CELLULAR COMPARTMENT, CD4+ T MEMORY CELLS (TM), WHICH CAN READILY RECOGNIZE MICROBIAL ANTIGENS FOR A QUICK AND ROBUST RESPONSE. FOR THIS PURPOSE, WE WILL UTILIZE A NEWLY CHARACTERIZED MURINE MODEL OF RECURRENT PD AND A METHOD FOR GINGIVAL T-CELL ENRICHMENT. IN THIS MODEL, TIME IS A CRITICAL VARIABLE FOR RECOGNIZING DISEASE INITIATION, RESOLUTION, AND RECURRENCE. BY ESTABLISHING THIS MODEL, WE WILL BE ABLE TO DEFINE A “RECOVERED” BASELINE INSTEAD OF A “HEALTHY” BASELINE STATE, WHICH IS MORE CLINICALLY ACCURATE. ALSO, IT ALLOWS DEFINING HOW MULTIPLE RELAPSE EPISODES INCREMENTALLY IMPACT THE COMPLEX PERIODONTALLY-ASSOCIATED IMMUNE NETWORK. THE PRELIMINARY STUDIES REVEALED THAT CD4+ TM DEVELOPED IN THE GINGIVA SOON AFTER BIRTH DUE TO INCREASING ENVIRONMENTAL ANTIGENIC EXPOSURE. WHILE THE MICE FULLY RECOVERED FROM LIGATURE-INDUCED PD, THE BONE LOSS RATE ACCELERATED DURING PD RECURRENCE. MOST INTRIGUINGLY, CD4+ TM SUBTYPES ARE ENRICHED IN THE GINGIVA DURING PD RECURRENCE. GINGIVAL CD4+ TISSUE-RESIDENT MEMORY CELLS INDUCED BONE LOSS WHEN CIRCULATING T CELL INFILTRATION WAS INHIBITED. BASED ON THESE FINDINGS, WE WILL TEST THE HYPOTHESIS THAT THE PD-INDUCED GENERATION AND PERSISTENCE OF CD4+ TM SUBTYPES DETERMINE PD RECURRENCE. THUS, TARGETED DEPLETION OF THESE CELLS BY INHIBITING METABOLIC CHECKPOINTS WILL ENHANCE IMMUNE-REGULATORY RESPONSES AND INHIBIT PD RECURRENCE. THERE ARE THREE INTERCONNECTED BUT INDEPENDENT AIMS: 1) TO IDENTIFY THE SPATIOTEMPORAL DEVELOPMENT OF CD4+ TM CELLS IN PERIODONTAL TISSUES. HERE, THE FOCUS WILL BE ON UNDERSTANDING THE CONSTITUTIVE AND PATHOLOGIC GENERATION OF CD4+ TM CELLS TO DETERMINE THEIR ROLES IN THE GINGIVAL IMMUNE LANDSCAPE. 2) TO ELUCIDATE PD-INDUCED CD4+ TM PATHOGENICITY. HERE, WE WILL EVALUATE WHICH CD4+ TM SUBTYPES RETAIN PRO-INFLAMMATORY AND OSTEOCLASTOGENIC PROGRAMMING AFTER PD. WE WILL MAKE TRANSCRIPTOMIC COMPARISONS BETWEEN CD4+ TM SUBTYPES AND ASSESS THE FINDINGS WITH EX VIVO FUNCTIONAL ASSAYS TO VALIDATE THEIR PATHOGENIC POTENTIAL. 3) TO DEVELOP AN INTERVENTION STRATEGY FOR CD4+ TM DEPLETION TO PREVENT PD RECURRENCE. HERE, WE WILL EMPLOY A METABOLIC-BASED STRATEGY TO SELECTIVELY DEPLETE PD-INDUCED CD4+ TM AND ENHANCE REGULATORY T CELLS ENRICHMENT TO PREVENT PD RECURRENCE. THE INDEPENDENCE AWARD (PAR-22-041) WILL FOSTER MY INDEPENDENT RESEARCH GROWTH AND ALLOW ME TO ACCOMPLISH MY LONG-TERM CAREER GOAL TO BECOME A PRODUCTIVE, INDEPENDENT TRANSLATIONAL SCIENTIST IN PERIODONTOLOGY AND RELATED FIELDS FOCUSED ON CHRONIC DISEASES’ RECURRENCE.

BIOMIMETIC GROWTH OF ENAMEL-LIKE HIERARCHICAL STRUCTURES

MICROCT CORE FACILITY IN THE FORSYTH INSTITUTE

NEW TECHNOLOGIES TO IDENTIFY NOVEL PERIODONTAL PATHOGENS

RECOMBINANT STREPTOCOCCUS MITIS VACCINE TECHNOLOGY AGAINST HIV AND OTHER DISEASES

PROTEIN-MINERAL INTERACTIONS DURING INITIAL STAGES OF ENAMEL FORMATION

TISSUE TRANSGLUTAMINASE DOMAINS THAT COMPLEX WITH BACTERIA

JEOL JSM-6510LV SCANNING ELECTRON MICROSCOPE

FORSYTH STUDENT SCHOLARS SUMMER INTERNSHIPS IN ORAL HEALTH RESEARCH - ORAL DISEASES DISPROPORTIONATELY IMPACT COMMUNITIES WITH LIMITED ACCESS TO HEALTHCARE. US SCHOOL SYSTEMS HAVE SYSTEMIC LIMITATIONS WHICH LEAD TO DIFFERING OUTCOMES FOR STUDENTS BASED ON OPPORTUNITY, WHICH HAVE PROFOUND AND LONG-LASTING CONSEQUENCES THAT CONTINUE TO HAVE PROFOUND IMPACTS ON OUR NATIONAL POTENTIAL. SINCE 1993, THE ADA FORSYTH STUDENT SCHOLARS (FSS) PROGRAM HAS PREPARED HIGH SCHOOL STUDENTS FOR SCIENCE AND RESEARCH PROGRAMS IN COLLEGE. OUR PROGRAM AIMS TO: 1) PRESENT SCIENCE, SCIENTISTS, AND ORAL HEALTH PROVIDERS AS MULTI-FACETED, ADVANCED AND ACCESSIBLE, 2) INCREASE BASIC RESEARCH UNDERSTANDING AND THE CAREER OPPORTUNITIES IT CREATES, AND 3) FOSTER INCREASED COMFORT LEVELS IN APPROACHING SCIENCE- AND ORAL HEALTH-RELATED CONCEPTS AND ACTIVITIES. THE FSS PROGRAM IS COMPRISED OF A MENTORED RESEARCH PROJECT AND TRAINING MODULES ON LABORATORY RESEARCH SKILLS, SCIENCE AND ORAL HEALTH SEMINARS, AND COLLEGE/CAREER READINESS SKILLS. UNIQUE AND INNOVATIVE ATTRIBUTES OF THE FSS HIGH SCHOOL PROGRAM INCLUDE INDIVIDUALIZED MENTORING, STUDENT INVOLVEMENT IN CUTTING-EDGE RESEARCH, TECHNOLOGY DEVELOPMENT TRAINING, SOFT SKILLS DEVELOPMENT, AND NETWORKING OPPORTUNITIES WITHIN BOSTON AND WITH LOCAL UNIVERSITY TO EMPHASIZE PUBLIC HEALTH NEEDS. THE FSS PROGRAM CURRICULUM IS UPDATED YEARLY FOR EVIDENCE-BASED IMPROVEMENT TO OPTIMIZE LEARNING OUTCOMES FOR OUR STUDENTS, USING INDIVIDUAL NEEDS ASSESSMENTS AND PROGRAM EVALUATIONS FROM BOTH STUDENTS AND MENTORS. THE LONG-TERM GOAL IS TO PROVIDE OPPORTUNITIES THAT HELP GROW THE FUTURE ORAL HEALTH RESEARCH WORKFORCE. AT THE SAME TIME, THE PROJECT HELPS TO RAISE ORAL HEALTH AWARENESS IN OUR COMMUNITIES, HELPING THE NEXT GENERATION OF DENTIST-SCIENTIST RESEARCHERS DEVELOP CRITICAL MENTORING SKILLS. APPLICANTS ARE RECRUITED TO THE FSS PROGRAM THROUGH OUTREACH EVENTS, VISITS TO HIGH SCHOOLS IN UNDER-RESOURCED MASSACHUSETTS COMMUNITIES, AND PARTNERSHIPS WITH COMMUNITY AND EDUCATION-BASED ORGANIZATIONS. OF THE ALMOST 60% OF FSS ALUMNI FOLLOWED IN 30 YEARS OF PROGRAMMING, 86% ARE CURRENTLY WORKING IN SCIENCE, AND 53% HAVE COMPLETED GRADUATE, MEDICAL, DENTAL, OR PROFESSIONAL SCHOOL. WHILE HAVING MANY OF THE SAME COMPONENTS, THE INCREASED RIGOR OF THE UNDERGRADUATE PROGRAM WILL HELP PREPARE STUDENTS FOR FURTHER STUDY IN PUBLIC HEALTH AND/OR THE SCIENCES. THIS PROPOSAL SEEKS FUNDS FOR TEN HIGH SCHOOL STUDENTS IN THE FSS PROGRAM AND CONTINUES OUR TRACK RECORD OF INCREASING STUDENTS' READINESS FOR COLLEGE, PROFESSIONAL SCHOOL, AND THE ORAL HEALTH WORKFORCE.

CAREER: BIOPHYSICS OF FLATWORM REGENERATION

M1/M2 REGULATION OF INFLAMMATION IN T2D-ASSOCIATED PERIODONTITIS

MOLECULAR AND PATHOGENIC STUDY OF AN ORAL TM7 STRAIN, THE FIRST CULTIVATED ULTRA-SMALL BACTERIUM

REGULATION OF CHEMR23 IN RESOLUTION OF INFLAMMATION

FUNCTIONAL ANALYSIS OF NHA-OC/NHA2 IN VIVO

ROLE OF INTERLEUKIN-22 IN THE SALIVARY GLAND DISORDER IN AUTOIMMUNE SJ?GREN'S SYNDROME

OPTIMIZED SECOND-GENERATION RECOMBINANT MYCOBACTERIA VACCINE VECTORS AGAINST HIV

POLYMICROBIAL DYNAMICS IN TRANSFER OF VANCOMYCIN RESISTANCE TO MRSA

INHIBITION OF P. GINGIVALIS BIOFILM FORMATION BY A STREPTOCOCCAL EFFECTOR PROTEIN

INVESTIGATING THE FUNCTION OF FIMBRIAE-FORMING LIPOPROTEIN IN PORPHYROMONAS GINGIVALIS - PORPHYROMONAS GINGIVALIS (PG) IS A HIGHLY PROTEOLYTIC GRAM-NEGATIVE ANAEROBE IMPLICATED IN PERIODONTAL DISEASE, WHICH IS ONE OF THE MOST COMMON CHRONIC BIOFILM-BASED INFECTIONS THAT RESULT IN DESTRUCTION OF HARD AND SOFT TISSUES OF THE PERIODONTIUM AND, ULTIMATELY, IN LOSS OF TEETH. ALTHOUGH PG HAS BEEN HISTORICALLY CLASSIFIED AS NONMOTILE, IT WAS RECENTLY SHOWN THAT FIMBRIATED STRAINS ARE CAPABLE OF SURFACE TRANSLOCATION WHEN SANDWICHED BETWEEN SOFT AGAR AND A GLASS OR PLASTIC SURFACE. THROUGH GENETIC STUDIES, IT WAS DETERMINED THAT THE TYPE IX SECRETION SYSTEM (T9SS) AND T9SS CARGO ARE INTEGRAL TO PG’S MIGRATION BEHAVIOR. OUR WORKING MODEL IS THAT DURING THE EARLY STAGES OF SURFACE TRANSLOCATION, T9SS CARGO PROTEINS WHICH ARE KNOWN TO BE TRANSPORTED TO THE CELL SURFACE AND RELEASED INTO THE ENVIRONMENT ON OUTER MEMBRANE VESICLES (OMVS) MODIFY THE SURROUNDINGS AND THIS PROMOTES MIGRATION. HOWEVER, THE COOPERATION AND FUNCTION OF THE VARIOUS OMV CARGO PROTEINS DURING THE TRANSITION TO SURFACE TRANSLOCATION IS STILL UNKNOWN. IN A TRANSCRIPTOME ANALYSIS, PG1881, A PREDICTED FIMBRIAE- FORMING LIPOPROTEIN, WAS ONE OF THE MOST HIGHLY UPREGULATED GENES DURING THE INITIAL STAGES OF SURFACE TRANSLOCATION, YET ITS FUNCTION IS NOT CLEAR. IMPORTANTLY, PG1881 HAS BEEN SHOWN TO BE ENRICHED ON SPHINGOLIPID (SL) CONTAINING OMVS. PREDICTED POST-TRANSLATIONAL MODIFICATIONS OF PG1881 INCLUDE PALMITOYLATION, PROVIDING A LINK TO SLS, AS WELL AS CITRULLINATION BY PPAD (PORPHYROMONAS PEPTIDYLARGININE DEIMINASE) WHICH CONVERTS L- ARGININE RESIDUES TO L-CITRULLINE WITHIN PEPTIDES. IT WAS PREVIOUSLY SHOWN THAT CITRULLINATION BY PPAD PROMOTES OMV BIOGENESIS AS WELL AS SURFACE TRANSLOCATION. THEREFORE, THIS STUDY AIMS TO INVESTIGATE THE FUNCTION OF PG1881 DURING THE EARLY STAGES OF SURFACE TRANSLOCATION. MY PRELIMINARY DATA EXAMINING OMVS FROM SURFACE TRANSLOCATING CELLS LACKING PG1881 REVEALED DISTINCT PROPERTIES VIA FLUORESCENCE AND TRANSMISSION ELECTRON MICROSCOPY. THEREFORE, THE CENTRAL HYPOTHESIS IS THAT PG1881 INFLUENCES THE BIOGENESIS AND PROPERTIES OF OMVS, IN PARTICULAR THE PROTEIN CARGO CARRIED ON OMVS, WHICH IMPACTS THE INITIAL STAGES OF SURFACE TRANSLOCATION. THIS HYPOTHESIS WILL BE TESTED THROUGH TWO SPECIFIC AIMS: 1) CHARACTERIZE PG1881 AND 2) INVESTIGATE THE FUNCTION OF PG1881 IN THE CONTEXT OF SURFACE TRANSLOCATION. IN AIM 1, PALMITOYLATION OF PG1881 WILL BE CONFIRMED BY USING MY PG1881 POLYCLONAL ANTIBODY TO PERFORM IMMUNOPRECIPITATION FOLLOWED BY MASS SPECTROMETRY. CITRULLINATION OF PG1881 WILL BE CONFIRMED USING RECOMBINANT PG1881 AS A SUBSTRATE FOR A COLORIMETRIC ASSAY THAT MEASURES CITRULLINATION. IN AIM 2, THE SPATIAL LOCALIZATION OF PG1881 ON SURFACE TRANSLOCATING CELLS WILL BE DETERMINED USING MY POLYCLONAL PG1881 ANTIBODY. I WILL COMPARE GINGIPAINS PROTEIN LEVELS AND ENZYMATIC ACTIVITY FROM SURFACE TRANSLOCATING CELLS IN THE PARENT STRAIN AND PG1881 DELETION MUTANT. THE EXPERIMENTS OUTLINED ABOVE WILL GIVE INSIGHT ON A NEW ASPECT OF PG PHYSIOLOGY AND CHARACTERIZE A PREDICTED FIMBRIAE-FORMING LIPOPROTEIN ASSOCIATED WITH KNOWN VIRULENCE DETERMINANTS INCLUDING OMVS AND T9SS. ULTIMATELY, THIS RESEARCH PROPOSAL IS THE BASIS OF A DOCTORAL DISSERTATION AND WILL ENHANCE THE TRAINING OF A STUDENT WITH AN INTEREST IN ORAL HEALTH RESEARCH.

ORAL MICROBIOME: BEYOND BACTERIA - THIS R13 PROPOSAL SEEKS SUPPORT FOR FORSYTH SYMPOSIUM 2021 “ORAL MICROBIOME: BEYOND BACTERIA” AT THE FORSYTH INSTITUTE ON OCTOBER 28-29, 2021. THE OVERALL OBJECTIVE OF THE SYMPOSIUM IS TO PROMOTE BETTER UNDERSTANDING OF ORAL MICROBIOME FROM A HOLISTIC PERSPECTIVE. ORAL MICROBIOME IS COMPOSED OF DIVERSE BACTERIAL SPECIES, FUNGAL AND VIRAL (BACTERIOPHAGE) COMPONENTS, AS WELL AS RECENTLY IDENTIFIED NANO-SIZED PARASITIC CANDIDATE PHYLA RADIATION (CPR) ORGANISMS. EFFECTIVE MANAGEMENT OF ORAL MICROBIOME-RELATED DISEASES CALLS FOR A MORE COMPREHENSIVE UNDERSTANDING OF THE IMPACT OF THE ORAL MICROBIOTA ON HOST HEALTH AND DISEASE, WHICH REQUIRES A HOLISTIC VIEW OF INTRA- AND CROSS-KINGDOM INTERACTIONS AMONG DIFFERENT MEMBERS OF THE ORAL MICROBIOME (INCLUDING BACTERIA, FUNGI, PHAGES AND CPR ORGANISMS), AS WELL AS MICROBIAL-HOST INTERACTION. THE SYMPOSIUM WILL BRING TOGETHER A GROUP OF SCIENTISTS WITH DIVERSE RESEARCH BACKGROUNDS IN ORAL MICROBIOME (INCLUDING ORAL BACSTERIOME, MYCOBIOME, PHAGEOME AND CPR) AND DIFFERENT CAREER STAGES TO SHARE THEIR RESEARCH AND IDEAS. THE SYMPOSIUM WILL INCLUDE POSTER PRESENTATIONS BY ATTENDEES AND TALKS BY: ANN GRIFFEN, MATTHEW M. RAMSEY, HYUN (MICHEL) KOO, JILL BANFIELD, MIRCEA PODAR, JEFF MCLEAN, XUESONG HE, KATHRYN KAUFFMAN, DAVID MOYES, MAHMOUD GHANNOUM, PATRICIA DIAZ, JESSICA MARK WELCH, YANG-YU LIU, AND ANNA EDLUND. ANOTHER KEY OBJECTIVE IS TO IDENTIFY BIOLOGICAL QUESTIONS AND TECHNICAL ROADBLOCKS THAT MUST BE OVERCOME TO ACHIEVE A MORE COMPREHENSIVE UNDERSTANDING OF THE IMPACT OF THE ORAL MICROBIOTA ON HOST HEALTH AND DISEASE FROM A HOLISTIC POINT OF VIEW. MEANWHILE, THE SYMPOSIUM WILL PROMOTE NEW COLLABORATIVE RESEARCHES WHICH WILL EXPAND FROM BACTERIAL INTERSPECIES INTERACTIONS TO INTER-KINGDOM INTERACTIONS WHICH INCLUDE BACTERIA, CPR, FUNGI, AND PHAGES. IT WILL ALSO PROVIDE A GREAT OPPORTUNITY FOR TRAINEES TO INTERACT WITH EXPERTS IN THE FIELD AND PROMOTE THEIR RESEARCH. MORE IMPORTANTLY, BY ENCOURAGING FEMALE TRAINEES AND TRAINEES WITH DISABILITIES OR OF MINORITY GROUPS, THE SYMPOSIUM AIMS TO PROMOTE THE ENGAGEMENT OF UNDERREPRESENTED GROUPS IN DENTAL RESEARCH. THE CONFERENCE WILL CREATE A COMMUNITY OF INVESTIGATORS WHO HAVE PREVIOUSLY RESIDED IN SCIENTIFIC SILOS THAT DID NOT MEET OR COMMUNICATE WITH EACH OTHER: ORAL BACTERIOME, MYCOBIOME, PHAGEOME AND CPR. THESE INVESTIGATORS WILL NOW COMMUNICATE AND HOPEFULLY DEVELOP COLLABORATIVE RESEARCH TO ACHIEVE A FUNDAMENTAL UNDERSTANDING OF ORAL MICROBIOME FROM A HOLISTIC PERSPECTIVE, WHICH WILL ULTIMATELY IMPROVE OUR ABILITY TO DIAGNOSE AND TREAT MICROBIAL INFECTIONS.

THE UNCULTIVABLE BACTERIA

2023 FORSYTH SYMPOSIUM: THE ORAL MICROBIOME: PAST, PRESENT, AND FUTURE - ABSTRACT THIS R13 PROPOSAL SEEKS SUPPORT FOR THE 2023 FORSYTH SYMPOSIUM ENTITLED “THE ORAL MICROBIOME: PAST, PRESENT, AND FUTURE,” WHICH IS PROPOSED TO TAKE PLACE AT THE FORSYTH INSTITUTE ON SEPTEMBER 14-15, 2023, WITH A PRE-CONFERENCE WORKSHOP ON PRESENTATION SKILLS FOR TRAINEES ON SEPTEMBER 13. THE OVERALL OBJECTIVES OF THE SYMPOSIUM ARE TO CATALYZE INNOVATIVE THINKING ABOUT AREAS WHERE FOCUSED BASIC RESEARCH ON THE ORAL MICROBIOME WILL LEAD TO IMPROVEMENTS IN HUMAN HEALTH; TO PROMOTE COLLABORATION AMONG RESEARCHERS USING NEW APPROACHES TO ADVANCE THE FIELD OF ORAL MICROBIAL ECOLOGY; AND TO HONOR AND CELEBRATE THE PIONEERING WORK OF FLOYD DEWHIRST AND BRUCE PASTER ON THE ORAL MICROBIOME, TO TRANSMIT THEIR KNOWLEDGE AND INSPIRE A YOUNGER GENERATION TO CONTINUE THEIR LEGACY OF INNOVATIVE SCIENCE. AN IMPORTANT GOAL IS TO PROVIDE THE OPPORTUNITY FOR TRAINEES TO INTERACT WITH EXPERTS IN THE FIELD AND TO ENCOURAGE FULL PARTICIPATION OF PERSONS WHO ARE TRADITIONALLY UNDER-REPRESENTED IN SCIENCE, INCLUDING WOMEN, MEMBERS OF MINORITY GROUPS, AND PERSONS WITH DISABILITIES. THE SYMPOSIUM WILL INCLUDE POSTER PRESENTATIONS BY ATTENDEES AND TALKS BY SUSAN BULLMAN, OTTO CORDERO, KATHERINE LEMON, TAMI LIEBERMAN, DENNIS MANGAN, DIANNE NEWMAN, FRANK NICHOLS, JESSICA SCOFFIELD, AND WILLIAM WADE. SCIENTIFIC QUESTIONS THAT WILL BE ADDRESSED BY THE SYMPOSIUM INCLUDE: WHAT ARE THE RULES GOVERNING COLONIZATION OF MICROBES AT DIFFERENT SITES WITHIN THE MOUTH AND WHAT GOVERNS WHETHER ORAL MICROBES COLONIZE ELSEWHERE IN THE BODY? HOW CAN THE STUDY OF BOTH ORAL AND NON-ORAL MICROBIAL COMMUNITIES ILLUMINATE ORAL MICROBIAL ECOLOGY? HOW DO ORAL BACTERIA INFLUENCE SYSTEMIC HEALTH AND DISEASE? THE SYMPOSIUM WILL ALSO SEEK TO BRING TOGETHER ACADEMIC SCIENTISTS WITH LEADERS FROM INDUSTRY AND VENTURE CAPITAL, TO BRAINSTORM ABOUT WHAT FOUNDATIONAL EXPERIMENTS COULD LAY THE GROUNDWORK FOR THERAPEUTIC MANIPULATION OF THE ORAL MICROBIOME. IF THIS CONFERENCE SUCCEEDS AS INTENDED, IT WILL ENABLE INDIVIDUAL INVESTIGATORS TO FORM NEW HIGH-QUALITY COLLABORATIONS AND DEVELOP INNOVATIVE GRANT PROPOSALS TO CARRY OUT FUNDAMENTAL RESEARCH THAT WILL ELUCIDATE THE MECHANISMS AND CONSEQUENCES OF ORAL MICROBIOME COLONIZATION, TAXON-TAXON INTERACTION, AND THE SYSTEMIC EFFECTS OF ORAL MICROBIOMES. EVEN MORE SIGNIFICANTLY, WE HOPE THAT THIS CONFERENCE WILL INSPIRE AND SUPPORT NEW FUNDING OPPORTUNITY ANNOUNCEMENTS BY FUNDING AGENCIES, AND NEW COLLABORATIONS BETWEEN BUSINESS AND ACADEMIC RESEARCHERS TO DRIVE INNOVATION IN ORAL MICROBIOME RESEARCH AND ITS CONTRIBUTION TO OVERALL HUMAN HEALTH AND WELL-BEING.

SPATIAL STATISTICS FOR ANALYSIS OF ORAL MICROBIAL COMMUNITY ORGANIZATION