Project Grant F31AI202997

TRYPANOSOMA CRUZI FLAGELLAR POCKET MORPHOGENESIS DURING CELL AND LIFE CYCLE TRANSITIONS - PROJECT SUMMARY TRYPANOSOMA CRUZI IS THE INFECTIOUS AGENT OF CHAGAS DISEASE, ENDEMIC TO SOUTH AND CENTRAL AMERICA. RECENTLY, CHAGAS DISEASE HAS BEEN PROPOSED TO BE ENDEMIC TO THE UNITED STATES, DUE TO GROWING CONCERNS OF U.S. CASES WITH NO KNOWN TRAVEL HISTORY AND EXPANSION OF VECTOR HABITATS. DESPITE AN ESTIMATED 6-7 MILLION PEOPLE CURRENTLY BEING AFFECTED WORLDWIDE, CHAGAS DISEASE IS STILL UNDER- DIAGNOSED AND LACKS THERAPEUTIC TREATMENT OPTIONS AND DIAGNOSTIC TOOLS. THROUGHOUT ITS CELL AND LIFE CYCLE, A PARASITE MUST UNDERGO DRASTIC MORPHOLOGICAL TRANSITIONS TO BECOME PRE-ADAPTED FOR SUCCESS IN DIVERSE ENVIRONMENTS. A KEY FEATURE CONTROLLING HOW A CELL INTERACTS WITH ITS SURROUNDINGS IS THE FLAGELLAR POCKET (FP), THE SITE WHERE THE SINGLE FLAGELLUM EXITS THE CELL BODY. THE FP PLAYS IMPORTANT ROLES IN EVASION OF THE HOST IMMUNE SYSTEM AND ENDO- AND EXOCYTOSIS OF MATERIAL IN AND OUT OF THE CELL. THE PROPOSED STUDY AIMS TO EXPLORE FP BIOGENESIS DURING T. CRUZI CELL DIVISION AS WELL AS HOW THE FP AND ASSOCIATED CYTOSKELETAL STRUCTURES ARE REARRANGED THROUGHOUT THE PARASITE'S LIFE CYCLE. IN AIM 1 WE WILL USE ULTRASTRUCTURE EXPANSION MICROSCOPY TO CHARACTERIZE FLAGELLAR POCKET ASSOCIATED CYTOSKELETAL STRUCTURE (FPACS) BIOGENESIS IN WILDTYPE PARASITES. THEN WE WILL DESCRIBE FPACS DUPLICATION DURING DEPLETION OF HOOK COMPLEX PROTEIN MORN1 AND FP COLLAR PROTEIN BILBO AS WELL AS INVESTIGATE THE ROLE OF THE HIGHLY CONSERVED EUKARYOTIC KINASE, TCPLK, IN THIS PROCESS. IN AIM 2 WE WILL AGAIN USE ULTRASTRUCTURE EXPANSION MICROSCOPY TO DEFINE FPACS, THIS TIME DURING METACYCLOGENESIS, THE PROCESS BY WHICH AN INSECT-RESIDENT PARASITE BECOMES HUMAN INFECTIOUS. FINALLY, WE WILL USE OUR LAB'S NOVEL LIVE-CELL IMAGING TECHNIQUE TO ELUCIDATE THE DYNAMICS OF FP REPOSITIONING DURING METACYCLOGENESIS. THE FINDINGS OF THIS PROPOSED WORK WILL REVEAL INSIGHTS INTO HOW THESE INFECTIOUS PARASITES REPLICATE AND COULD THEREFORE HELP TO IDENTIFY NOVEL CANDIDATES FOR DRUG DESIGN. FURTHER, THIS STUDY WILL IDENTIFY MECHANISMS OF CELL REARRANGEMENT DURING THE T. CRUZI INFECTIOUS CYCLE AND COULD AID IN THE DEVELOPMENT OF STRATEGIES TO BLOCK TRANSMISSION FROM INSECT VECTOR TO HUMAN, THUS PREVENTING THE SPREAD OF THIS NEGLECTED TROPICAL DISEASE.