Primary Research Focus
Research in the Pilaz Lab focuses on cellular and molecular mechanisms regulating the production, migration and survival of neurons in the context of brain development in health and disease. Four lines of investigation are currently followed.
Missense mutations or deletion of the ZBTB7A gene lead to macrocephaly (larger brain) and intellectual disability but the mechanisms underlying these phenotypes are completely unknown. Since ZBTB7A encodes a transcription factor regulating genes essential for proliferation and cell fate in various lineages, we predicted that ZBTB7A regulates brain size and function through the regulation of neural stem cell proliferation. Our current data from mouse models supports this hypothesis, and we are now dissecting the underlying molecular mechanisms, with a particular focus on how ZBTB7A interactions with the Nucleosome Remodeling and Deacetylation complex (NuRD) influence neural stem cells.
Pathogenic variants in MAP1B were recently associated with Periventricular Nodular Heterotopia (PVNH), a neuron migration deficit. We aim to understand the mechanism of MAP1B-PVNH and characterize the defects of affected neurons. To do this, we use CRISPR in the embryonic mouse cortex to introduce MAP1B-PVNH variants in neurons and neural progenitors. This leads to a significant migration delay and produces a truncated MAP1B protein. Overexpression of the same variant does not recapitulate this migration delay, indicating that this does not act as a dominant negative. Future directions include further characterization of the MAP1B-PVNH model, including cytoskeletal abnormalities and the mechanism underlying migration deficits.
Beta-propeller associated neurodegeneration (BPAN) is a devastating rare disease affecting the central nervous system and resulting from mutations in WDR45. Patients present with seizures, intellectual disabilities, delayed speech, ataxia and autism spectrum disorders, all starting in infancy and early childhood. Eventually, BPAN culminates with massive neurodegeneration in early adulthood. There is no cure for BPAN and standard treatment is symptomatic as the disease progresses. In order to push BPAN research forward, we generated a new BPAN mouse model inspired by a patient mutation (c52C>T), resulting in early premature truncation of WIPI4 at amino acid residue 18. Characterization of this mouse shows behavioral deficits as early as 3 months, as well anatomical deficits. Current work focuses on the molecular mechanisms by which Wdr45 loss leads to these defects.
We love to develop new tools to give researchers the means to push the limits of knowledge. When it comes to brain development, our go-to method is in utero electroporation, allowing us to introduce DNA, RNA and proteins, into live embryonic mouse brains. We couple in utero electroporation with CRISPR to alter the genome of neural stem cells and their progeny using a technique we developed called Breasi-CRISPR. We mainly use Breasi-CRISPR to tag endogenous proteins and visualize them within isolated cells, both live and fixed. Current efforts try to leverage Breasi CRISPR to tag mRNAs instead of proteins.
About the Pilaz Lab
Lab Projects and News
Epigenetic mechanisms regulating neuron production during cortical development
A myriad of molecular pathways are known to control precursor cell proliferation. Specifically, cycling precursor cells utilize epigenetic mechanisms to fine-tune the expression of proliferation genes. While recent studies have indicated that epigenetic regulation by chromatin accessibility factors is critical in neural stem cells, the complete mechanism of such regulatory factors remains to be discovered. In this study, we investigate the molecular mechanism by which an unconventional disease-linked epigenetic regulator impacts neural stem cells proliferation, and thus the development of the cerebral cortex.
Role of RNA trafficking and local translation in neural stem cells
Using live imaging in embryonic brain tissue, we recently visualized RNA being trafficked in neural stem cells. In these cells, we made the surprising observation that RNA can be actively transported over long distances to locally produce proteins far from their nucleus and cell body. We used RNA-immunoprecipitation to discover 115 transported RNAs, but the role of this mechanism remains enigmatic. In this project, we focus on three autism-linked transported RNAs to understand the function of this mechanism, and how its disruption may lead to disease.
Meet the Pilaz Lab
Pratiksha KC, MS
Pratiksha KC joined the Pilaz Lab in 2020 to assist the team in research related to transcriptional and post-transcriptional mechanisms that drive neuron and neural stem cell biology during embryonic brain development.
Pratiksha earned her master’s degree in molecular biology from South Dakota State University. She has three years of experience in molecular biology techniques, such as molecular cloning.
Claire is an MD-PhD student who joined the Pilaz lab in February 2021 to complete her dissertation. Prior to joining the lab, Claire obtained her BA in cellular neuroscience from Colgate University. At Colgate she studied the molecular mechanisms of dendritic arborization. Following graduation, Claire spent two years at the NIH studying neuronal signal transduction in the context of somatosensation and neural development. In the Pilaz lab, she studies how patient variants in MAP1B lead to periventricular nodular heterotopia (PVNH), a rare neurodevelopmental disease. She uses techniques such as in utero electroporation to study PVNH and prenatal brain development. After graduation, she plans to pursue a career as a physician scientist specializing in pediatrics and medical genetics.
Brandon joined the Pilaz lab in 2019 as Graduate Assistant while seeking his PhD through the Basic Biomedical Sciences program at the University of South Dakota Sanford School of Medicine. His work focuses on pediatric neurological disorders resulting from neurodevelopmental defects and neurodegenerative decline. His work expands our knowledge of how the brain develops and how the cells of the brain maintain themselves through the lifetime of the individual.
Krishna joined the Pilaz Lab after gaining skills working in industry and receiving a master's degree from the University of South Florida in Pharmaceutical Sciences.