High throughput genomic edition to investigate neurodevelopmental disorders using isogenic cellular models

• Grant number: 21/09089-0
• Support Opportunities: Research Grants - Young Investigators Grants
• Start date: February 01, 2023
• End date: January 31, 2028
• Field of knowledge: Biological Sciences - Genetics - Molecular Genetics and Genetics of Microorganisms
• Principal Investigator: Mariana Moysés Oliveira
• Grantee: Mariana Moysés Oliveira
• Host Institution: Associação Fundo de Incentivo à Pesquisa (AFIP). São Paulo , SP, Brazil
• Associated researchers: Andrea Parolin Jackowski ; Daniela Santoro Rosa ; Gabriel Natan de Souza Pires ; Gustavo José da Silva Pereira ; Lygia da Veiga Pereira ; Maria Isabel de Souza Aranha Melaragno ; Monica Levy Andersen ; Sandra Doria Xavier ; Sergio Tufik ; Síntia Iole Nogueira Belangero
• Associated research grant(s): 23/01515-6 - Identification of molecular profiles in the Rare Genetic Syndrome associated with SYNGAP1 in isogenic neuronal models, AP.R
• Associated scholarship(s): 24/13762-0 - Genetic editing in induced pluripotent stem cells for the production of isogenic neural models with variants in POGZ for the study of neurodevelopmental disorders, BP.MS; 24/13810-5 - Evaluation of the endocannabinoid system in isogenic neural models of neurodevelopmental disorders associated with sleep alterations, BP.DD; 24/00578-7 - The genes behind rarities: a collective construction about the bases of rare genetic diseases, BP.JC; + associated scholarships 24/00579-3 - The genes behind rarities: a collective construction about the bases of rare genetic diseases, BP.JC; 23/14935-3 - Analysis of genetic editing and expansion efficiency on isogenic cellular models subjected to CRISPR/Cas9 methodologies, BP.IC; 23/01662-9 - High throughput genomic edition to investigate neurodevelopmental disorders associated with CREBBP using isogenic cellular models, BP.IC - associated scholarships

Abstract

The genetic architecture of neurodevelopmental disorders (NDD) includes rare, highly penetrant mutations and common variants of smaller effect. De novo, rare loss-of-function (LoF) and missense mutations in evolutionarily constrained genes are the strongest known risk factors for NDD, and these genes are particularly enriched in two main pathways: (1) neuronal communication and (2) regulation of gene expression, i.e., chromatin regulators and transcription factors. Functional analyses suggest that these two pathways are interconnected during neurodevelopment; however, their mechanisms of convergence are unknown. Moreover, the relative impact of LoF variants compared to missense mutations is poorly understood. In this proposal, we will leverage a CRISPR-based high throughput genomic engineering platform for parallelized genome edition in human induced pluripotent stem cell (hiPSC) lines to introduce LoF and missense perturbations on NDD-associated genes related to transcription regulation. hiPSC isogenic cellular models will be generated and differentiated in neural stem cells and glutamatergic cortical neurons to interrogate the molecular consequences the genomic editions created. We will explore direct, or proximal, dysregulation of critical neuronal genes following the LoF of these transcription regulators (Aim 1), and then compare the relative impact of patient-specific missense variants vs. gene deletions (Aim 2). These hiPSC-derived neuronal lineages will have their transcriptome and chromatin accessibility profile defined by RNA-seq and Assay for Transposase-Accessible Chromatin (ATAC)-seq to infer the molecular consequences followed by the introduction of these genomic variants. This study will thus leverage new technologies, neuronal models, and complementary datasets to identify the nodes of coalescence in molecular signatures of NDD, which may correspond to druggable targets in future studies. (AU)

Scientific publications

(20)

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

GUERREIRO, P.; MOYSES-OLIVEIRA, M.; KLOSTER, A.; CUNHA, L.; DECONTO, T. BASSANI; MOSINI, A. CRISTINA; MARQUEZINI, B. PEREIRA; PASCHALIDIS, M.; GALLEGO ADAMI, L. NAYARA; ANDERSEN, M. L.; et al. SLEEP DISTURBANCES ASSOCIATED WITH DEAF1 PATHOGENIC VARIANTS. Sleep Medicine, v. 115, p. 2-pg., 2024-02-01. (20/13467-8, 21/09089-0)

KLOSTER, ANNA K.; ADAMI, LUANA N. G.; MOYSES-OLIVEIRA, MARIANA; HACHUL, HELENA; ANDERSEN, MONICA L.; TUFIK, SERGIO. Exploring the Link between Premature Ovarian Insufficiency, Insomnia and Circadian Pathways. JORNAL BRASILEIRO DE REPRODUCAO ASSISTIDA, v. 28, n. 4, p. 23-pg., 2024-10-01. (23/11995-5, 21/09089-0, 20/13467-8)

KLOSTER, A.; MOYSES-OLIVEIRA, M. M. -O.; ADAMI, L. N. G.; HACHUL, H.; ANDERSEN, M. L.; TUFIK, S.. EXPLORING THE LINK BETWEEN PREMATURE OVARIAN INSUFFICIENCY, INSOMNIA AND CIRCADIAN PATHWAYS. Sleep Medicine, v. 115, p. 1-pg., 2024-02-01. (21/09089-0, 20/13467-8)

MOSINI, AMANDA CRISTINA; ADAMI, LUANA NAYARA GALLEGO; VALLIM, JULIA RIBEIRO DA SILVA; MOYSES-OLIVEIRA, MARIANA; POYARES, DALVA; ANDERSEN, MONICA L.; TUFIK, SERGIO. Leptin moderates the relationship between sleep quality and memory function: A population-based study. Sleep Medicine, v. 117, p. 6-pg., 2024-03-26. (23/09760-0, 21/09089-0, 20/13467-8)

MOYSES-OLIVEIRA, MARIANA; ZAMARIOLLI, MALU; TEMPAKU, PRISCILA F.; GALDUROZ, JOSE CARLOS FERNANDES; ANDERSEN, MONICA L.; TUFIK, SERGIO. Shared genetic mechanisms underlying association between sleep disturbances and depressive symptoms. Sleep Medicine, v. 119, p. 9-pg., 2024-04-18. (21/09089-0, 20/13467-8)

MOYSES-OLIVEIRA, MARIANA; FAVILLA, BIANCA PEREIRA; MELARAGNO, MARIA ISABEL; TUFIK, SERGIO. X-Chromosome Dependent Differences in the Neuronal Molecular Signatures and Their Implications in Sleep Patterns. SLEEP MEDICINE CLINICS, v. 18, n. 4, p. 11-pg., 2023-10-27. (21/09089-0)

MOSINI, AMANDA CRISTINA; MOYSES-OLIVEIRA, MARIANA; DE ARAUJO, JESSICA NAYARA GOES; GUERREIRO, PEDRO; CUNHA, LAIS; ZAMARIOLLI, MALU; XAVIER, SANDRA DORIA; BALBUENO, BIANCA; DE MELLO, CLAUDIA BERLIM; MOREIRA, GUSTAVO ANTONIO; et al. Sleep complaints in individuals with SYNGAP1-associated syndrome. Sleep Medicine, v. 126, p. 8-pg., 2024-12-28. (21/09089-0, 20/13467-8, 24/05023-3, 23/14935-3, 23/09760-0)

CUNHA, L.; MARQUEZINI, B. P.; MOYSES-OLIVEIRA, M.; KLOSTER, A.; DECONTO, T. B.; MOSINI, A. C.; GUERREIRO, P.; PASCHALIDIS, M.; ADAMI, L. N. G.; ANDERSEN, M. L.; et al. THE RELATIONSHIP BETWEEN CREBBP VARIANTS AND INSOMNIA: FROM RUBINSTEIN-TAYBI SYNDROME INTO ENERGY METABOLISM. Sleep Medicine, v. 115, p. 2-pg., 2024-02-01. (21/09089-0, 20/13467-8)

CUNHA, LAIS; PASCHALIDIS, MAYARA; MOYSES-OLIVEIRA, MARIANA; MARQUEZINI, BRUNA PEREIRA; DECONTO, TAIS BASSANI; GUERREIRO, PEDRO; KLOSTER, ANNA; MOSINI, AMANDA CRISTINA; ADAMI, LUANA NAYARA GALLEGO; ANDERSEN, MONICA L.; et al. The relationship between neurodevelopmental transcriptional programs and insomnia: From Rubinstein-Taybi syndrome into energy metabolism. Sleep Medicine, v. 112, p. 3-pg., 2023-10-05. (21/09089-0, 20/13467-8)

PASCHALIDIS, M.; MOYSES-OLIVEIRA, M.; KLOSTER, A.; MARQUEZINI, B.; CUNHA, L.; ADAMI, L. N. G.; GUERREIRO, P.; MOSINI, A. C.; ANDERSEN, M. L.; TUFIK, S.. NEUROTRANSMITTER REGULATION AS COMMON PATHWAYS BETWEEN SLEEP PHENOTYPES, RESTLESS LEG SYNDROME AND TOURETTE SYNDROME. Sleep Medicine, v. 115, p. 2-pg., 2024-02-01. (21/09089-0, 20/13467-8)

MOYSES-OLIVEIRA, M.; ANDERSEN, M. L.; TUFIK, S.. X-CHROMOSOME FUNCTIONAL DOSAGE IMPACT NEURONAL MOLECULAR SIGNATURES AND CIRCADIAN REGULATION. Sleep Medicine, v. 115, p. 2-pg., 2024-02-01. (21/09089-0, 20/13467-8)

MOYSES-OLIVEIRA, MARIANA; PASCHALIDIS, MAYARA; SOUZA-CUNHA, LAIS A. A.; ESTEVES-GUERREIRO, PEDRO A. A.; ADAMI, LUANA NAYARA GALLEGO; KLOSTER, ANNA K. K.; MOSINI, AMANDA C. C.; MOREIRA, GUSTAVO A. A.; DORIA, SANDRA; TEMPAKU, PRISCILA F. F.; et al. Genetic basis of sleep phenotypes and rare neurodevelopmental syndromes reveal shared molecular pathways. Journal of Neuroscience Research, v. N/A, p. 10-pg., 2023-02-15. (21/09089-0)

ADAMI, L.; OLIVEIRA, M.; CUNHA, L.; TUFIK, S.; ANDERSEN, M.. LIPID METABOLISM AND NEUROMUSCULAR JUNCTION AS COMMON PATHWAYS UNDERLYING THE GENETIC BASIS OF ERECTILE DYSFUNCTION AND OBSTRUCTIVE SLEEP APNEA. Sleep Medicine, v. 115, p. 1-pg., 2024-02-01. (21/09089-0, 20/13467-8)

ADAMI, LUANA N. G.; MOYSES-OLIVEIRA, MARIANA; SOUZA-CUNHA, LAIS AMANDA; VASCO, MATHEUS BRANDAO; TUFIK, SERGIO; ANDERSEN, MONICA L.. Lipid metabolism and neuromuscular junction as common pathways underlying the genetic basis of erectile dysfunction and obstructive sleep apnea. INTERNATIONAL JOURNAL OF IMPOTENCE RESEARCH, v. 36, n. 6, p. 7-pg., 2023-11-21. (23/11995-5, 21/09089-0, 20/13467-8)

MARQUEZINI, BRUNA PEREIRA; MOYSES-OLIVEIRA, MARIANA; KLOSTER, ANNA; CUNHA, LAIS; DECONTO, TAIS BASSANI; MOSINI, AMANDA CRISTINA; GUERREIRO, PEDRO; PASCHALIDIS, MAYARA; ADAMI, LUANA NAYARA GALLEGO; ANDERSEN, MONICA LEVY; et al. Exploring the molecular pathways linking sleep phenotypes and POGZ-associated neurodevelopmental disorder. JOURNAL OF MEDICAL GENETICS, v. 61, n. 6, p. 4-pg., 2024-02-13. (21/09089-0, 20/13467-8)

XAVIER, SANDRA D.; MOYSES-OLIVEIRA, MARIANA; KLOSTER, ANNA KAROLINY; BALBUENO, BIANCA; TUFIK, SERGIO; ANDERSEN, MONICA L.. Evaluation of the Sleep Profile in Children With Cri du Chat Syndrome: A Pilot Study. PEDIATRIC NEUROLOGY, v. 156, p. 2-pg., 2024-04-30. (21/09089-0, 20/13467-8)

MARQUEZINI, B. P.; MOYSES-OLIVEIRA, M.; KLOSTER, A.; CUNHA, L.; DECONTO, T. B.; MOSINI, A. C.; GUERREIRO, P.; PASCHALIDIS, M.; ADAMI, L. N. G.; ANDERSEN, M. L.; et al. EXPLORING THE MOLECULAR PATHWAYS LINKING SLEEP PHENOTYPES AND POGZ-ASSOCIATED NEURODEVELOPMENTAL DISORDERS. Sleep Medicine, v. 115, p. 2-pg., 2024-02-01. (21/09089-0, 20/13467-8)

MOSINI, A. C.; MOYSES-OLIVEIRA, M.; DECONTO, T. B.; PEREIRA MARQUEZINI, B.; GUERREIRO, P.; CUNHA, L.; KLOSTER, A.; PASCHALIDIS, M.; GALLEGO ADAMI, L. N.; ANDERSEN, M. L.; et al. MUTATIONAL MECHANISMS RELATED TO SLEEP DISTURBANCE IN PATIENTS WITH SYNGAP1-ASSOCIATED SYNDROME. Sleep Medicine, v. 115, p. 1-pg., 2024-02-01. (21/09089-0, 20/13467-8)

ADAMI, LUANA N. G.; MOYSES-OLIVEIRA, MARIANA; TUFIK, SERGIO; ANDERSEN, MONICA L.. Kinesin binding as a shared pathway underlying the genetic basis of male factor infertility and insomnia. F&S SCIENCE, v. 5, n. 3, p. 7-pg., 2024-08-01. (23/11995-5, 21/09089-0, 20/13467-8)

MOYSES-OLIVEIRA, MARIANA; ADAMI, LUANA N. G.; GUERREIRO, PEDRO; MOSINI, AMANDA CRISTINA; KLOSTER, ANNA; CUNHA, LAIS; PASCHALIDIS, MAYARA; MARQUEZINI, BRUNA PEREIRA; PIRES, GABRIEL N.; TEMPAKU, PRISCILA F.; et al. Endocrine and Epigenetic Regulation as Common Pathways Underlying the Genetic Basis of Sleep Traits and Longevity. REJUVENATION RESEARCH, v. 26, n. 5, p. 8-pg., 2023-10-01. (21/09089-0)