Biochemistry, Genetics and Molecular Biology, Genetics, Cellular and Molecular Neuroscience
34
Scopus Publications
Scopus Publications
“SHANK3 deficiency alters early progenitor dynamics and reveals shared pathways with neurodegeneration” Elisa Varella-Branco, Elizabeth Shephard, Victor H. C. Toledo, Igor C. Ramos, Ellen C. M. Lacerda, Laura L. M. Carvalho, Marcella A. Fiuza, Mayara Paschalidis, Claudia I. S. Costa, Ana C. S. Girardi, Ana C. V. Krepischi, Erasmo B. Casella, Guilherme Polanczyk, Karina Griesi-Oliveira, Fabio Papes, Lucas Alvizi, Gerson S. Kobayashi, Maria Rita dos Santos e Passos Bueno Molecular Psychiatry, 2026 Phelan-McDermid Syndrome (PMS), primarily linked to SHANK3 haploinsufficiency, presents with complex neurodevelopmental features, including developmental regression, whose underlying mechanisms are poorly understood. This study investigated the impact of SHANK3 disruption across multiple levels, from gene expression in patient-derived iPSC neurons to in vivo brain network activity. RNA-sequencing of iPSC-derived neurons from PMS patients with SHANK3 disruption only (n = 9) and controls (n = 7) revealed dysregulation in differential gene expression and co-expression modules linked to cell cycle, RNA metabolism, and metabolic pathways in SHANK3 -mutated neurons. All modules were correlated with PMS regression and enriched for genes implicated in neurodevelopmental or neurodegenerative disorders, such as autism, ADHD, and Alzheimer’s disease. At the cellular level, SHANK3 -mutated cultures exhibited increased proliferation of neural progenitors and intermediate progenitor markers. Differentiated neurons showed reduced morphological complexity, specific changes in postsynaptic marker density and puncta size, and electrophysiological characteristics suggestive of neuronal hyperexcitability. Electroencephalography (EEG) in a PMS patient cohort (n = 20) compared to controls (n = 30) demonstrated hyperconnectivity and excessive high-frequency oscillations, suggesting altered neural network dynamics. In summary, the use of different analytical approaches suggested that SHANK3 haploinsufficiency disrupts neurodevelopmental trajectories and revealed that regression in PMS may share common genes and pathways with neurodegeneration. We also characterized molecular and neurophysiological markers that can be useful in therapeutic protocols for PMS.
Chemical transdifferentiation of somatic cells to neural cells: a systematic review Paulo Victor Visintin, Bruna Lancia Zampieri, Karina Griesi-Oliveira Einstein Sao Paulo Brazil, 2024 INTRODUCTION: Transdifferentiation is the conversion of a specific somatic cell into another cell type, bypassing a transient pluripotent state. This implies a faster method to generate cells of interest with the additional benefit of reduced tumorigenic risk for clinical use. OBJECTIVE: We describe protocols that use small molecules as direct conversion inducers, without the need for exogenous factors, to evaluate the potential of cell transdifferentiation for pharmacological and clinical applications. METHODS: In this systematic review, using PRISMA guidelines, we conducted a personalized search strategy in four databases (PubMed, Scopus, Embase, and Web Of Science), looking for experimental works that used exclusively small molecules for transdifferentiation of non-neural cell types into neural lineage cells. RESULTS: We explored the main biological mechanisms involved in direct cell conversion induced by different small molecules used in 33 experimental in vitro and in vitro transdifferentiation protocols. We also summarize the main characteristics of these protocols, such as the chemical cocktails used, time for transdifferentiation, and conversion efficiency. CONCLUSION: Small molecules-based protocols for neuronal transdifferentiation are reasonably safe, economical, accessible, and are a promising alternative for future use in regenerative medicine and pharmacology.
Omics profile of iPSC-derived astrocytes from Progressive Supranuclear Palsy (PSP) patients Felipe G. Ravagnani, Hellen P. Valerio, Jersey H.S. Maués, Arthur N. de Oliveira, Renato D. Puga, Karina Griesi-Oliveira, Fabíola R. Picosse, Henrique B. Ferraz, Rodrigo R. Catharino, Graziella E. Ronsein, Patrícia de Carvalho Aguiar Parkinsonism and Related Disorders, 2023
Rare CACNA1H and RELN variants interact through mTORC1 pathway in oligogenic autism spectrum disorder André Luíz Teles e Silva, Talita Glaser, Karina Griesi-Oliveira, Juliana Corrêa-Velloso, Jaqueline Yu Ting Wang, Gabriele da Silva Campos, Henning Ulrich, Andrea Balan, Mehdi Zarrei, Edward J. Higginbotham, Stephen W. Scherer, Maria Rita Passos-Bueno, Andrea Laurato Sertié Translational Psychiatry, 2022 Oligogenic inheritance of autism spectrum disorder (ASD) has been supported by several studies. However, little is known about how the risk variants interact and converge on causative neurobiological pathways. We identified in an ASD proband deleterious compound heterozygous missense variants in the Reelin (RELN) gene, and a de novo splicing variant in the Cav3.2 calcium channel (CACNA1H) gene. Here, by using iPSC-derived neural progenitor cells (NPCs) and a heterologous expression system, we show that the variant in Cav3.2 leads to increased calcium influx into cells, which overactivates mTORC1 pathway and, consequently, further exacerbates the impairment of Reelin signaling. Also, we show that Cav3.2/mTORC1 overactivation induces proliferation of NPCs and that both mutant Cav3.2 and Reelin cause abnormal migration of these cells. Finally, analysis of the sequencing data from two ASD cohorts—a Brazilian cohort of 861 samples, 291 with ASD; the MSSNG cohort of 11,181 samples, 5,102 with ASD—revealed that the co-occurrence of risk variants in both alleles of Reelin pathway genes and in one allele of calcium channel genes confer significant liability for ASD. Our results support the notion that genes with co-occurring deleterious variants tend to have interconnected pathways underlying oligogenic forms of ASD.
Molecular Characterization of a First-in-Human Clinical Response to Nimesulide in Acute Myeloid Leukemia Victória Tomaz, Karina Griesi-Oliveira, Renato D. Puga, Bruno J. Conti, Fabio P. S. Santos, Nelson Hamerschlak, Paulo V. Campregher Frontiers in Oncology, 2022 Acute myeloid leukemia (AML) is a hematologic malignancy associated with high morbidity and mortality. Here we describe a case of a patient with AML who presented a partial response after utilization of the non-steroidal anti-inflammatory drug nimesulide. The response was characterized by complete clearance of peripheral blood blasts and an 82% decrease of bone marrow blasts associated with myeloblast differentiation. We have then shown that nimesulide induces in vitro cell death and cell cycle arrest in all AML cell lines (HL-60, THP-1, OCI-AML2, and OCI-AML3). Weighted Correlation Network Analysis (WGCNA) of serial whole-transcriptome data of cell lines treated with nimesulide revealed that the sets of genes upregulated after treatment with nimesulide were enriched for genes associated with autophagy and apoptosis, and on the other hand, the sets of downregulated genes were associated with cell cycle and RNA splicing. Serial transcriptome of bone marrow patient sample confirmed the upregulation of genes associated with autophagy after the response to nimesulide. Lastly, we demonstrated that nimesulide potentiates the cytotoxic in vitro effect of several Food and Drug Administration (FDA)-approved chemotherapy drugs used in AML, including cytarabine.
Interleukin-17a Induces Neuronal Differentiation of Induced-Pluripotent Stem Cell-Derived Neural Progenitors From Autistic and Control Subjects Ana Karolyne Santos Gomes, Rafaelly Mayara Dantas, Bruno Yukio Yokota, André Luiz Teles e Silva, Karina Griesi-Oliveira, Maria Rita Passos-Bueno, Andréa Laurato Sertié Frontiers in Neuroscience, 2022 Prenatal exposure to maternal immune activation (MIA) has been suggested to increase the probability of autism spectrum disorder (ASD). Recent evidence from animal studies indicates a key role for interleukin-17a (IL-17a) in promoting MIA-induced behavioral and brain abnormalities reminiscent of ASD. However, it is still unclear how IL-17a acts on the human developing brain and the cell types directly affected by IL-17a signaling. In this study, we used iPSC-derived neural progenitor cells (NPCs) from individuals with ASD of known and unknown genetic cause as well as from neurotypical controls to examine the effects of exogenous IL-17a on NPC proliferation, migration and neuronal differentiation, and whether IL-17a and genetic risk factors for ASD interact exacerbating alterations in NPC function. We observed that ASD and control NPCs endogenously express IL-17a receptor (IL17RA), and that IL-17a/IL17RA activation modulates downstream ERK1/2 and mTORC1 signaling pathways. Exogenous IL-17a did not induce abnormal proliferation and migration of ASD and control NPCs but, on the other hand, it significantly increased the expression of synaptic (Synaptophysin-1, Synapsin-1) and neuronal polarity (MAP2) proteins in these cells. Also, as we observed that ASD and control NPCs exhibited similar responses to exogenous IL-17a, it is possible that a more inflammatory environment containing other immune molecules besides IL-17a may be needed to trigger gene-environment interactions during neurodevelopment. In conclusion, our results suggest that exogenous IL-17a positively regulates the neuronal differentiation of human NPCs, which may disturb normal neuronal and synaptic development and contribute to MIA-related changes in brain function and behavior.
Neuroprogenitor Cells From Patients With TBCK Encephalopathy Suggest Deregulation of Early Secretory Vesicle Transport Danielle de Paula Moreira, Angela May Suzuki, André Luiz Teles e Silva, Elisa Varella-Branco, Maria Cecília Zorél Meneghetti, Gerson Shigeru Kobayashi, Mariana Fogo, Merari de Fátima Ramires Ferrari, Rafaela Regina Cardoso, Naila Cristina Vilaça Lourenço, Karina Griesi-Oliveira, Elaine Cristina Zachi, Débora Romeo Bertola, Karina de Souza Weinmann, Marcelo Andrade de Lima, Helena Bonciani Nader, Andrea Laurato Sertié, Maria Rita Passos-Bueno Frontiers in Cellular Neuroscience, 2022 Biallelic pathogenic variants in TBCK cause encephaloneuropathy, infantile hypotonia with psychomotor retardation, and characteristic facies 3 (IHPRF3). The molecular mechanisms underlying its neuronal phenotype are largely unexplored. In this study, we reported two sisters, who harbored biallelic variants in TBCK and met diagnostic criteria for IHPRF3. We provided evidence that TBCK may play an important role in the early secretory pathway in neuroprogenitor cells (iNPC) differentiated from induced pluripotent stem cells (iPSC). Lack of functional TBCK protein in iNPC is associated with impaired endoplasmic reticulum-to-Golgi vesicle transport and autophagosome biogenesis, as well as altered cell cycle progression and severe impairment in the capacity of migration. Alteration in these processes, which are crucial for neurogenesis, neuronal migration, and cytoarchitecture organization, may represent an important causative mechanism of both neurodevelopmental and neurodegenerative phenotypes observed in IHPRF3. Whether reduced mechanistic target of rapamycin (mTOR) signaling is secondary to impaired TBCK function over other secretory transport regulators still needs further investigation.
Complement c4 is reduced in ipsc-derived astrocytes of autism spectrum disorder subjects Fernanda Mansur, André Luiz Teles e Silva, Ana Karolyne Santos Gomes, Juliana Magdalon, Janaina Sena de Souza, Karina Griesi-Oliveira, Maria Rita Passos-Bueno, Andréa Laurato Sertié International Journal of Molecular Sciences, 2021 In recent years, accumulating evidence has shown that the innate immune complement system is involved in several aspects of normal brain development and in neurodevelopmental disorders, including autism spectrum disorder (ASD). Although abnormal expression of complement components was observed in post-mortem brain samples from individuals with ASD, little is known about the expression patterns of complement molecules in distinct cell types in the developing autistic brain. In the present study, we characterized the mRNA and protein expression profiles of a wide range of complement system components, receptors and regulators in induced pluripotent stem cell (iPSC)-derived neural progenitor cells, neurons and astrocytes of individuals with ASD and neurotypical controls, which constitute in vitro cellular models that recapitulate certain features of both human brain development and ASD pathophysiology. We observed that all the analyzed cell lines constitutively express several key complement molecules. Interestingly, using different quantification strategies, we found that complement C4 mRNA and protein are expressed in significantly lower levels by astrocytes derived from ASD individuals compared to control astrocytes. As astrocytes participate in synapse elimination, and diminished C4 levels have been linked to defective synaptic pruning, our findings may contribute to an increased understanding of the atypically enhanced brain connectivity in ASD.
TRPC channels and mental disorders Karina Griesi-Oliveira, Angela May Suzuki, Alysson Renato Muotri Advances in Experimental Medicine and Biology, 2017
Modeling non-syndromic autism and the impact of TRPC6 disruption in human neurons K Griesi-Oliveira, A Acab, A R Gupta, D Y Sunaga, T Chailangkarn, X Nicol, Y Nunez, M F Walker, J D Murdoch, S J Sanders, T V Fernandez, W Ji, R P Lifton, E Vadasz, A Dietrich, D Pradhan, H Song, G-l Ming, X Gu, G Haddad, M C N Marchetto, N Spitzer, M R Passos-Bueno, M W State, A R Muotri Molecular Psychiatry, 2015
Overexpression of KLC2 due to a homozygous deletion in the non-coding region causes SPOAN syndrome Uirá S. Melo, Lucia I. Macedo-Souza, Thalita Figueiredo, Alysson R. Muotri, Joseph G. Gleeson, Gabriela Coux, Pablo Armas, Nora B. Calcaterra, João P. Kitajima, Simone Amorim, Thiago R. Olávio, Karina Griesi-Oliveira, Giuliana C. Coatti, Clarissa R.R. Rocha, Marinalva Martins-Pinheiro, Carlos F.M. Menck, Maha S. Zaki, Fernando Kok, Mayana Zatz, Silvana Santos Human Molecular Genetics, 2015
HTR1B and HTR2C in autism spectrum disorders in Brazilian families G.M. Orabona, K. Griesi-Oliveira, E. Vadasz, V.L.S. Bulcão, V.N.V.O. Takahashi, E.S. Moreira, M. Furia-Silva, A.M.S. Ros-Melo, F. Dourado, R. Matioli, P. Otto, M.R. Passos-Bueno Brain Research, 2009
