Project Grant F30MH142021

MAPPING CONVERGENCE IN AUTISM AND SCHIZOPHRENIA: HIGH-THROUGHPUT CHARACTERIZATION OF GLUTAMATERGIC NEURONS HARBORING PERTURBATIONS ACROSS 25 DISTINCT GENES - PROJECT SUMMARY: NEURODEVELOPMENTAL AND NEUROPSYCHIATRIC DISORDERS (NPD) LIKE AUTISM SPECTRUM DISORDER (ASD) AND SCHIZOPHRENIA (SCZ), ARE HIGHLY HERITABLE LIFELONG POLYGENIC DISORDERS. DESPITE EXTENSIVE PROGRESS IN IDENTIFYING GENETIC RISK GENES, THEIR BIOLOGICAL FUNCTION AND IMPLICATIONS ON NEURODEVELOPMENT REMAIN UNKNOWN AND CONVENTIONAL APPROACHES FOCUS ON SINGLE-GENE MANIPULATION, LIMITING THE ABILITY TO IDENTIFY CONVERGENCE AND CAUSING DELAYS IN STUDYING THE POLYGENICITY OF THESE DISORDERS. ADVANCED MOLECULAR TECHNIQUES LIKE CRISPR/CAS9 AND HUMAN PLURIPOTENT STEM CELL (HPSC) MODELS OFFER A WAY TO ANALYZE MULTIPLE GENES' MOLECULAR PHENOTYPES SIMULTANEOUSLY. LEVERAGING HPSC-DERIVED GLUTAMATERGIC NEURONS, A KEY CELL TYPE IN ASD AND SCZ BIOLOGY, THIS STUDY AIMS TO CHARACTERIZE THE IMPACT OF KNOCKING OUT 25 NPD GENES TO BETTER UNDERSTAND DISEASE MECHANISMS, INTRA AND INTER-DISEASE CONVERGENCE, AND POSSIBLE AVENUES FOR DOWNSTREAM THERAPEUTIC TARGETS. IN AIM 1 I WILL INVESTIGATE HOW EACH HOMOZYGOUS AND HETEROZYGOUS KO AFFECTS NEURAL DEVELOPMENT AND MORPHOLOGY. THIS INCLUDES ASSESSING THE IMPACT EACH KO HAS ON NEURALIZATION, AND DEVELOPMENTAL TRAJECTORIES DURING DIFFERENTIATION AS WELL AS PHENOTYPING CHANGES IN NEURONAL MORPHOLOGY AT MATURATION. IN AIM 2 I WILL PERFORM BULK RNA SEQUENCING AND HIGH SENSITIVITY MASS SPECTROMETRY AT TWO DEVELOPMENTAL TIME POINTS FOR EACH LINE, FOLLOWED BY A COMPREHENSIVE ANALYSIS COMPARING ACROSS METHODOLOGIES, ZYGOSITY, GENES, AND DISEASE SUBCLUSTERS, TO REVEAL SHARED AND UNIQUE SIGNATURES WHILE MAPPING BIOLOGICAL NETWORKS ACROSS NPD. TOGETHER, THESE AIMS WILL ELUCIDATE A PATH TO BETTER STUDY POLYGENIC DISORDERS, INSIGHT MECHANISMS FOR HOW NPD GENETIC RISK CONTRIBUTES TO NEURODEVELOPMENT AND PROVIDE POSSIBLE AVENUES FOR UNCOVERING THERAPEUTIC TARGETS BY IDENTIFYING CONVERGENCE ACROSS DISEASE BIOLOGY.