The NEUROSTAD project conducted under this MSCA Research Fellowship, has made significant progress in unravelling the role of Stau2 in neurogenesis and its implications in AD. The project was organized into six work packages (WPs), each contributing to the overall objectives.
WP1 included deleting STAU2 from hiPSC lines and investigating the impact of Stau2 depletion on neuron's development. The genome editing CRISPR/Cas9 system was employed for precise STAU2 knockout, resulting in a stable hiPSC cell line. The differentiation potential of these cells was assessed, revealing accelerated neurogenesis in STAU2 knockout conditions. This work represents a scientific milestone, providing a robust platform for exploring Stau2's role in neurogenesis.
WP2 comprised the single-cell library formation and sequencing, using Drop-seq for cell separation into oil vesicles. Despite a maternity leave pause, the deliverables for scRNA-seq libraries were successfully achieved. This package represents a crucial step, due that the rest of the project depended on the successful sequencing and on the quality of the data. The datasets obtained will be deposited for broader accessibility.
WP3 focused on analyzing scRNA-seq data from WP2, identifying mRNA candidates pivotal in neurogenesis and AD. The analysis revealed significant changes in STAU2 content at earlier stages of neurogenesis than previously described. Deliverables involving the analysis of RNA-seq datasets were achieved, providing a list of candidate genes associated with neurogenesis upon STAU2 knockout and in AD cells.
WP4 aimed to perform all the the experimental and functional analysis of selected targets identified in WP3. We optimized the transwell experiments to analyze changes in mRNA localization upon STAU2 KO and the APEX system to analyze the RNA-STAU2 protein complexes formed in each stage of neurogenesis. This work will continue even after the MSCA fellowship (ongoing collaboration between the Fellow and the Host lab).
WP5 and WP6 were about management, training and development. During the work on research projects, the Fellow also supervised a Erasmus+ Exchange Technician from the Gouti Lab at MDC (Berlin, Germany) and a Erasmus+ master student from Gratz University; and co-tutorized a Master Student from the University of Barcelona (UB). She has received training in single-cell library preparation and bioinformatics, hiPSC generation, maintenance, and characterization, and successful knock-outs by using CRISPR-Cas techniques (via collaboration during project in the Host lab). Furthermore, the Fellow got additional training in optical microscopy, flow cytometry analysis and how to give powerful presentations through IDIBELL Areté Program and Leadership and managerial skills through Marie Curie Alumni Association (MCAA) courses through Coursera. Management skills were further promoted collaborating closely with the post-award unit at IDIBELL, with constant communication with relevant departments, and utilization of the intranet for project details. Results of our research have been reflected in a scientific review (already accepted at Transcription Journal) and two forecoming papers (manuscripts in preparation). This work has been presented at three conferences (two via oral communication and one via poster communication). Moreover, this work was presented regularly at Department seminars at P-CMRC Department in IDIBELL and in the institutional fellows seminar. Moreover, the Fellow actively participated in science dissemination programs like INSPIRASTEAM and contributed to the MCAA newsletter, enhancing visibility in the scientific community Supervision and Training.
