Role of profilin1 in neocortical development and gyrification

• Applicants: Professorin Silvia Cappello, Ph.D.; Professor Dr. Marco Rust
• Subject Area: Developmental Neurobiology ; Molecular Biology and Physiology of Neurons and Glial Cells
• Term: from 2018 to 2023
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 391970916

• Final Report Year: 2023

Final Report Abstract

The mammalian brain owns two layered structures, the cerebral and the cerebellar cortex. While the cerebellar cortex is highly folded in all mammalian species, mammals are subdivided into lissencephalic and gyrencephalic species possessing either an unfolded or folded cerebrum, respectively. Current research focuses on investigating the mechanisms underlying cerebral and cerebellar folding and on elucidating the differences in cerebral cortex development between lissencephalic and gyrencephalic species. Overarching aim of this project was to elucidate the function of the actin regulator profilin1 in brain development. It was based on preliminary data in profilin1-KO (PFN1-KO) mice, in which increased numbers of proliferating cells outside the embryonic ventricular zone (VZ) was associated with the occurrence of mild cerebral folds. Our data in PFN1-KO mice led us propose a model in which profilin1-dependent actin assembly controls division of apical radial glia and thereby the fate of their progenies. Via this mechanism, profilin1 restricts VZ cell delamination and production of basal radial glia, and it thereby controls cerebral cortex development in mice. Besides this, we found impaired cerebellar folding in PFN1-KO mice. Interestingly, inactivation of the profilin1 interaction partner CAP1 caused a very similar defect in mouse cerebellar foliation. In CAP1-KO mice, cerebellar defects were associated with altered gene expression. In future studies, we will elucidate the molecular defects causing cerebellar changes in both mutant strains. Inspired by our mouse data demonstrating a role for PFN1 in restricting the generation of a greater diversity of neural progenitors, we tested whether it has acquired a similar function in human cerebrum. To assess this, we exploited IPS-derived human cerebral organoids. Unfortunately, it was not possible to generate IPS KO cells, probably because PFN1 is very important at the stem cell level. Since PFN1 overexpression led to premature cell death, we investigated the expression, localization and secretion of PFN1 and its homolog PFN2 in cerebral organoids. Finally, by performing interactome studies, we identified both as exclusive interacting partners of a pathological form of LGALS3BP, which caused changes in progenitor heterogeneity and alterations in the folding pattern of the patient's brain, suggesting a common mechanism regulating brain folding.

Publications

• Profilin1-Dependent F-Actin Assembly Controls Division of Apical Radial Glia and Neocortex Development. Cerebral Cortex, 30(6), 3467-3482.
  Kullmann, Jan A.; Meyer, Sophie; Pipicelli, Fabrizia; Kyrousi, Christina; Schneider, Felix; Bartels, Nora; Cappello, Silvia & Rust, Marco B.
  
• CAPt’n of Actin Dynamics: Recent Advances in the Molecular, Developmental and Physiological Functions of Cyclase-Associated Protein (CAP). Frontiers in Cell and Developmental Biology, 8.
  Rust, Marco B.; Khudayberdiev, Sharof; Pelucchi, Silvia & Marcello, Elena
  
• Cystatin B is essential for proliferation and interneuron migration in individuals with EPM 1 epilepsy. EMBO Molecular Medicine, 12(6).
  Di Matteo, Francesco; Pipicelli, Fabrizia; Kyrousi, Christina; Tovecci, Isabella; Penna, Eduardo; Crispino, Marianna; Chambery, Angela; Russo, Rosita; Ayo‐Martin, Ane Cristina; Giordano, Martina; Hoffmann, Anke; Ciusani, Emilio; Canafoglia, Laura; Götz, Magdalena; Di Giaimo, Rossella & Cappello, Silvia
  
• Functional Redundancy of Cyclase-Associated Proteins CAP1 and CAP2 in Differentiating Neurons. Cells, 10(6), 1525.
  Schneider, Felix; Metz, Isabell; Khudayberdiev, Sharof & Rust, Marco B.
  
• Mutual functional dependence of cyclase-associated protein 1 (CAP1) and cofilin1 in neuronal actin dynamics and growth cone function. Progress in Neurobiology, 202, 102050.
  Schneider, Felix; Duong, Thuy-An; Metz, Isabell; Winkelmeier, Jannik; Hübner, Christian A.; Endesfelder, Ulrike & Rust, Marco B.
  
• Disease association of cyclase-associated protein (CAP): Lessons from gene-targeted mice and human genetic studies. European Journal of Cell Biology, 101(2), 151207.
  Rust, Marco B. & Marcello, Elena
  
• Extracellular vesicle-mediated trafficking of developmental cues is altered during human brain disease. Cold Spring Harbor Laboratory.
  Pipicelli, Fabrizia; Forero, Andrea; Moser, Sylvain; Matteo, Francesco Di; Baumann, Natalia; Grätz, Christian; Pisfil, Mariano Gonzalez; Bekjarova, Zagorka; Pfaffl, Michael W.; Canafoglia, Laura; Pütz, Benno; Kielkowski, Pavel; Cernilogar, Filippo M.; Maccarrone, Giuseppina; Jabaudon, Denis; Giaimo, Rossella Di & Cappello, Silvia
  
• Non–cell-autonomous regulation of interneuron specification mediated by extracellular vesicles. Science Advances, 9(20).
  Pipicelli, Fabrizia; Baumann, Natalia; Di Giaimo, Rossella; Forero-Echeverry, Andrea; Kyrousi, Christina; Bonrath, Rebecca; Maccarrone, Giuseppina; Jabaudon, Denis & Cappello, Silvia