Huntington's disease (HD) is an inherited disease that causes degeneration of adult neurons in the striatum and later in the cortex. Although most studies have focused on adulthood, some evidence suggests that this disease might have a neurodevelopmental component. To try to address this question, we generated a protocol to derive 3D ventral organoids from isogenic control and pathologic hES cell line to mimic in-vitro the development of the ventral telencephalon. Immunocytochemistry analyses show correct identity acquisition, particularly if compared with dorsal protocols. These data are supported by single cell RNA-seq analyses that highlight the presence of ventral progenitors and GABAergic neurons with no dorsal subpopulations. Compared to controls, HD organoids show a 50% reduction in the number of regions that resemble the ventricular zone of the developing brain together with abnormalities in their polarity and compromised radial distribution of GSX2+ and MASH1+ progenitors. These early defects are combined with aberrant cell cycle regulation as shown by a 45% reduction in the number of KI67+ cells, a 95% increase in asymmetric horizontal divisions and a 15% increase in cell cycle exit ratio. Moreover, scanning electron microscopy (SEM) analyses reveal irregular neuronal architecture in HD organoids. Together our data suggest that the presence of mutant Huntingtin (mHTT) causes a developmental defect in the cellular organization of ventral organoids. This 3D model has therefore paved the way to a better understanding of the potential neurodevelopment aspects of this disease and may provide a platform to further screen and comprehend this complex disease. In line, we are now analyzing single cell data from HD organoids to unveil the transcriptional pathways affected by mHTT and in parallel we are setting up a co-culture system of control and HD cells to study cellular interactions between the two lines and to discriminate cell-autonomous versus non-autonomous defects.
MODELING HD AND HUMAN STRIATAL DEVELOPMENT USING 3D CULTURES / M. Galimberti ; scientific tutor: E. Cattaneo. - Milano : Università degli studi di Milano. Dipartimento di Bioscienze, 2020 May 04. ((32. ciclo, Anno Accademico 2019.
|Titolo:||MODELING HD AND HUMAN STRIATAL DEVELOPMENT USING 3D CULTURES|
|Data di pubblicazione:||4-mag-2020|
|Settore Scientifico Disciplinare:||Settore BIO/11 - Biologia Molecolare|
|Citazione:||MODELING HD AND HUMAN STRIATAL DEVELOPMENT USING 3D CULTURES / M. Galimberti ; scientific tutor: E. Cattaneo. - Milano : Università degli studi di Milano. Dipartimento di Bioscienze, 2020 May 04. ((32. ciclo, Anno Accademico 2019.|
|Appare nelle tipologie:||Tesi di dottorato|