Pilaz Lab

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.