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Investigation of degradation mechanisms of transcription factor 4 (TCF4), associated with autism spectrum disorders and schizophrenia

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Author(s):
Carolina Nunes Santo
Total Authors: 1
Document type: Master's Dissertation
Press: Campinas, SP.
Institution: Universidade Estadual de Campinas (UNICAMP). Instituto de Biologia
Defense date:
Examining board members:
Fabio Papes; Guilherme Oliveira Barbosa; Janaína Sena de Souza
Advisor: Mário Henrique Bengtson; Fabio Papes
Abstract

Transcription factor 4 (TCF4) is a basic helix-loop-helix protein highly expressed during neurodevelopment with an important role in neural progenitor cell maturation, neurogenesis and neuronal migration. Polymorphisms in TCF4 are associated with numerous neuropsychiatric disorders including bipolar disorder, depression, schizophrenia and autism. In addition, TCF4 haploinsufficiency leads to Pitt-Hopkins Syndrome (PTHS) development, a rare genetic disorder belonging to the autistic spectrum characterized by intellectual disability, severe motor delay, enteric nervous system dysfunction and specific facial dysmorphisms. The insufficient knowledge about the basic biology of TCF4 added to the poor understanding of the molecular mechanisms of the disease collaborate to the lack of specific therapies so far. Since TCF4 mutations in PTHS carriers are autosomal loss-of-function mutations, increasing the amount and/or activity of TCF4 in nervous system cells would be a possible therapeutic strategy for the correction or attenuation of this pathology. Thus, the identification of key enzymes of the TCF4 ubiquitin-proteasome-mediated protein degradation pathway represents a potential avenue for the identification of new therapeutic targets for PTHS and the future development of specific therapies. As E3 ligase enzymes dictate the specificity of target protein recognition and its tagging for degradation by the ubiquitin-proteasome pathway, we devised a strategy where we used a commercial CRISPR knockout library containing sgRNAs for all E3 ligases encoded in the human genome to infect neural cells stably expressing the accessory enzyme Cas9 and reporters for TCF4 quantification detectable by flow cytometry, with the aim to identify cells with altered TCF4 metabolism after CRISPR screening from reporter fluorescence and consequently identify the E3 ligase involved in TCF4 degradation (AU)

FAPESP's process: 21/10189-0 - Investigation of degradation mechanisms of Transcription Factor 4 (TCF4), associated with autism spectrum disorders and schizophrenia
Grantee:Carolina Nunes Santo
Support Opportunities: Scholarships in Brazil - Master