Define the mechanisms through which STK33 regulates multiciliated cells

About This Grant

PROJECT SUMMARY/ABSTRACT The following proposal outlines a 5-year career training plan that will prepare Dr. Andrew Berical to be an independent physician-scientist and leader in the field of airway epithelial functional genomics. Motile cilia are found throughout the human body, most notably on multiciliated cells (MCCs) in the conducting airway. Individuals with primary ciliary dyskinesia (PCD) have inherited variants in any one of more than 50 genes that regulate the structure or function of cilia, leading to a lifetime of chronic cough, recurrent infections and respiratory failure. Due to the complexity of the MCC molecular program and limited disease-relevant platforms, there are no targeted therapies available for PCD. An improved understanding of fundamental MCC biology and the availability of a human-based platform would have enormous implications for the PCD field. Dr. Berical’s long-term vision is to utilize pluripotent stem cell-based techniques to understand how specific genes regulate airway epithelial homeostasis and how gene variants lead to the initiation of airway diseases such as PCD, CF, asthma, COPD and IPF. Dr. Berical presents preliminary data suggesting a recently described serine-threonine kinase (STK33) has a fundamental role in the MCC developmental program. STK33 deletion results in 1) fewer MCCs, 2) fewer cilia per cell, 3) an abnormal ciliary structure and 4) reduced ciliary beat frequency. In this proposal, Dr. Berical aims to understand the mechanism by which STK33 effects the MCC molecular program to create this highly irregular phenotype. Leveraging key training opportunities through his collaborators and scientific advisory committee, he will 1) precisely characterize the STK33-dependent MCC defects using time course single cell RNA-sequencing to pinpoint when, during MCC differentiation, STK33 exerts its effect, 2) identify STK33 downstream targets and effector molecules and 3) determine the in vivo ramifications of STK33 loss on the engraftment, differentiation and function of airway epithelial cells. Following this investigation of the STK33-dependent regulation of MCC biology, Dr. Berical then expands these methods to probe the functions of a curated list of high priority ciliary kinases of unknown function. This work will provide much needed insight into the MCC molecular program and develop an essential platform for the interrogation of genes of unknown function in the airway epithelium, applicable to the genetically heterogeneous PCD, as well as other airway diseases. Dr. Berical has 80% protected time from his department to accomplish these aims under the guidance of his mentors Drs. Finn Hawkins and Darrell Kotton at the Center for Regenerative Medicine at Boston University/Boston Medical Center. He has assembled a remarkable team of advisors with diverse expertise to assist in his career development and scientific research. Dr. Berical details a comprehensive training plan that includes experiential training, didactic coursework, attendance and presentation at scientific meetings, preparation of manuscripts and acquiring additional grant support culminating in an R01. Dr. Berical has the commitment of his department to accomplish these goals and transition to an independent physician-scientist position by the end of the award.