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Article ; Online: Uncovering the bookshelves of CRISPR-based libraries: Advances and applications in cancer studies.

Quintero-Ruiz, Nathalia / Oliveira, Wesley de Lima / Esteca, Marcos Vinicius / Granato, Daniela Campos / Simabuco, Fernando Moreira

Critical reviews in oncology/hematology

2024 Volume 196, Page(s) 104287

Abstract: The advent of CRISPR/Cas9 technology has revolutionized the genome editing field. CRISPR-based libraries have become powerful tools for high-throughput functional genomics and genetic screening. CRISPR-based libraries can represent a powerful approach to ...

Abstract	The advent of CRISPR/Cas9 technology has revolutionized the genome editing field. CRISPR-based libraries have become powerful tools for high-throughput functional genomics and genetic screening. CRISPR-based libraries can represent a powerful approach to uncovering genes related to chemoresistance and therapy efficacy and to studying cancer cells' fitness. In this review, we conducted an extensive literature search and summarized multiple studies that utilized these libraries in both in vitro and in vivo research, emphasizing their key findings. We provide an overview of the design, construction, and applications of CRISPR-based libraries in different cancer-focused studies and discuss the different types of CRISPR-based libraries. We finally point out the challenges associated with library design, including guide RNA selection, off-target effects, and library complexity. This review provides an overview of the work conducted with CRISPR libraries in the search for new targets that could potentially assist in cancer therapy by contributing to functional approaches.
MeSH term(s)	Humans ; CRISPR-Cas Systems ; RNA, Guide, CRISPR-Cas Systems ; Gene Editing ; Gene Library ; Neoplasms/genetics ; Neoplasms/therapy
Chemical Substances	RNA, Guide, CRISPR-Cas Systems
Language	English
Publishing date	2024-02-10
Publishing country	Netherlands
Document type	Journal Article ; Review
ZDB-ID	605680-5
ISSN	1879-0461 ; 0737-9587 ; 1040-8428
ISSN (online)	1879-0461
ISSN	0737-9587 ; 1040-8428
DOI	10.1016/j.critrevonc.2024.104287
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Article ; Online: The respiratory syncytial virus M2-2 protein is targeted for proteasome degradation and inhibits translation and stress granules assembly.

Scudero, Orlando Bonito / Santiago, Verônica Feijoli / Palmisano, Giuseppe / Simabuco, Fernando Moreira / Ventura, Armando Morais

PloS one

2023 Volume 18, Issue 7, Page(s) e0289100

Abstract: The M2-2 protein from the respiratory syncytial virus (RSV) is a 10 kDa protein expressed by the second ORF of the viral gene M2. During infection, M2-2 has been described as the polymerase cofactor responsible for promoting genome replication, which ... ...

Abstract	The M2-2 protein from the respiratory syncytial virus (RSV) is a 10 kDa protein expressed by the second ORF of the viral gene M2. During infection, M2-2 has been described as the polymerase cofactor responsible for promoting genome replication, which occurs by the induction of changes in interactions between the polymerase and other viral proteins at early stages of infection. Despite its well-explored role in the regulation of the polymerase activity, little has been made to investigate the relationship of M2-2 with cellular proteins. A previous report showed poor recruitment of M2-2 to viral structures, with the protein being mainly localized to the nucleus and cytoplasmic granules. To unravel which other functions M2-2 exerts during infection, we performed proteomic analysis of co-immunoprecipitated cellular partners, identifying enrichment of proteins involved with regulation of translation, protein folding and mRNA splicing. In approaches based on these data, we found that M2-2 expression downregulates eiF2α phosphorylation and inhibits both translation and stress granules assembly. Finally, we also verified that M2-2 is targeted for proteasome degradation, being localized to granules composed of defective ribosomal products at the cytoplasm. These results suggest that besides its functions in the replicative complex, M2-2 may exert additional functions to contribute to successful RSV infection.
MeSH term(s)	Proteasome Endopeptidase Complex ; Proteomics ; Stress Granules ; Viral Proteins/genetics ; Respiratory Syncytial Virus, Human/genetics ; Virus Replication/physiology
Chemical Substances	Proteasome Endopeptidase Complex (EC 3.4.25.1) ; Viral Proteins
Language	English
Publishing date	2023-07-25
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2267670-3
ISSN	1932-6203 ; 1932-6203
ISSN (online)	1932-6203
ISSN	1932-6203
DOI	10.1371/journal.pone.0289100
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Article ; Online: Does a 217-km mountain ultramarathon affect the gut microbiota of a top 10 runner at the Brazil 135 Ultramarathon?

Saragiotto, Giulio Kai / de Oliveira, Luiz Felipe Valter / de Oliveira, Milena Merizzi / Simabuco, Fernando Moreira / Belli, Taisa / Antunes, Adriane Elisabete Costa

Scandinavian journal of medicine & science in sports

2023 Volume 33, Issue 12, Page(s) 2620–2622

MeSH term(s)	Humans ; Gastrointestinal Microbiome ; Brazil ; Running ; Athletes
Language	English
Publishing date	2023-10-06
Publishing country	Denmark
Document type	Letter
ZDB-ID	1077418-x
ISSN	1600-0838 ; 0905-7188
ISSN (online)	1600-0838
ISSN	0905-7188
DOI	10.1111/sms.14512
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Article: Sunflower and Palm Kernel Meal Present Bioaccessible Compounds after Digestion with Antioxidant Activity.

Bisinotto, Mariana Sisconeto / da Silva Napoli, Daniele Cristina / Simabuco, Fernando Moreira / Bezerra, Rosângela Maria Neves / Antunes, Adriane Elisabete Costa / Galland, Fabiana / Pacheco, Maria Teresa Bertoldo

Foods (Basel, Switzerland)

2023 Volume 12, Issue 17

Abstract: Sunflower ( ...

Abstract	Sunflower (
Language	English
Publishing date	2023-09-01
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2704223-6
ISSN	2304-8158
ISSN	2304-8158
DOI	10.3390/foods12173283
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Article ; Online: NEK6 Regulates Redox Balance and DNA Damage Response in DU-145 Prostate Cancer Cells.

Pavan, Isadora Carolina Betim / Basei, Fernanda Luisa / Severino, Matheus Brandemarte / Rosa E Silva, Ivan / Issayama, Luidy Kazuo / Mancini, Mariana Camargo Silva / Góis, Mariana Marcela / da Silva, Luiz Guilherme Salvino / Bezerra, Rosangela Maria Neves / Simabuco, Fernando Moreira / Kobarg, Jörg

Cells

2023 Volume 12, Issue 2

Abstract: NEK6 is a central kinase in developing castration-resistant prostate cancer (CRPC). However, the pathways regulated by NEK6 in CRPC are still unclear. Cancer cells have high reactive oxygen species (ROS) levels and easily adapt to this circumstance and ... ...

Abstract	NEK6 is a central kinase in developing castration-resistant prostate cancer (CRPC). However, the pathways regulated by NEK6 in CRPC are still unclear. Cancer cells have high reactive oxygen species (ROS) levels and easily adapt to this circumstance and avoid cell death by increasing antioxidant defenses. We knocked out the NEK6 gene and evaluated the redox state and DNA damage response in DU-145 cells. The knockout of NEK6 decreases the clonogenic capacity, proliferation, cell viability, and mitochondrial activity. Targeting the NEK6 gene increases the level of intracellular ROS; decreases the expression of antioxidant defenses (SOD1, SOD2, and PRDX3); increases JNK phosphorylation, a stress-responsive kinase; and increases DNA damage markers (p-ATM and γH2AX). The exogenous overexpression of NEK6 also increases the expression of these same antioxidant defenses and decreases γH2AX. The depletion of NEK6 also induces cell death by apoptosis and reduces the antiapoptotic Bcl-2 protein. NEK6-lacking cells have more sensitivity to cisplatin. Additionally, NEK6 regulates the nuclear localization of NF-κB2, suggesting NEK6 may regulate NF-κB2 activity. Therefore, NEK6 alters the redox balance, regulates the expression of antioxidant proteins and DNA damage, and its absence induces the death of DU-145 cells. NEK6 inhibition may be a new strategy for CRPC therapy.
MeSH term(s)	Male ; Humans ; Prostatic Neoplasms, Castration-Resistant/genetics ; Prostatic Neoplasms, Castration-Resistant/metabolism ; Reactive Oxygen Species/metabolism ; Cell Line, Tumor ; Antioxidants/metabolism ; NF-kappa B p52 Subunit/metabolism ; Oxidation-Reduction ; DNA Damage ; NIMA-Related Kinases/genetics ; NIMA-Related Kinases/metabolism
Chemical Substances	Reactive Oxygen Species ; Antioxidants ; NF-kappa B p52 Subunit ; NEK6 protein, human (EC 2.7.11.1) ; NIMA-Related Kinases (EC 2.7.11.1)
Language	English
Publishing date	2023-01-07
Publishing country	Switzerland
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2661518-6
ISSN	2073-4409 ; 2073-4409
ISSN (online)	2073-4409
ISSN	2073-4409
DOI	10.3390/cells12020256
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Article: Molecular mechanisms and pharmacological interventions in the replication cycle of human coronaviruses.

Simabuco, Fernando Moreira / Tamura, Rodrigo Esaki / Pavan, Isadora Carolina Betim / Morale, Mirian Galliote / Ventura, Armando Morais

Genetics and molecular biology

2020 Volume 44, Issue 1 Suppl 1, Page(s) e20200212

Abstract: SARS-CoV-2 (Severe Acute Respiratory Syndrome Coronavirus 2), as well as SARS-CoV from 2003 along with MERS-CoV from 2012, is a member of the Betacoronavirus genus of the Nidovirales order and is currently the cause of the pandemic called COVID-19 (or ... ...

Abstract	SARS-CoV-2 (Severe Acute Respiratory Syndrome Coronavirus 2), as well as SARS-CoV from 2003 along with MERS-CoV from 2012, is a member of the Betacoronavirus genus of the Nidovirales order and is currently the cause of the pandemic called COVID-19 (or Coronavirus disease 2019). COVID-19, which is characterized by cough, fever, fatigue, and severe cases of pneumonia, has affected more than 23 million people worldwide until August 25th, 2020. Here, we present a review of the cellular mechanisms associated with human coronavirus replication, including the unique molecular events related to the replication transcription complex (RTC) of coronaviruses. We also present information regarding the interactions between each viral protein and cellular proteins associated to known host-pathogen implications for the coronavirus biology. Finally, a specific topic addresses the current attempts for pharmacological interventions against COVID-19, highlighting the possible effects of each drug on the molecular events of viral replication. This review intends to aid future studies for a better understanding of the SARS-CoV-2 replication cycle and the development of pharmacological approaches targeting COVID-19.
Language	English
Publishing date	2020-11-23
Publishing country	Brazil
Document type	Journal Article
ZDB-ID	1445712-x
ISSN	1678-4685 ; 1415-4757
ISSN (online)	1678-4685
ISSN	1415-4757
DOI	10.1590/1678-4685-GMB-2020-0212
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Article ; Online: Effects of short-term endurance and strength exercise in the molecular regulation of skeletal muscle in hyperinsulinemic and hyperglycemic Slc2a4

Muñoz, Vitor Rosetto / Botezelli, José Diego / Gaspar, Rafael Calais / da Rocha, Alisson L / Vieira, Renan Fudoli Lins / Crisol, Barbara Moreira / Braga, Renata Rosseto / Severino, Matheus Brandemarte / Nakandakari, Susana Castelo Branco Ramos / Antunes, Gabriel Calheiros / Brunetto, Sérgio Q / Ramos, Celso D / Velloso, Lício Augusto / Simabuco, Fernando Moreira / de Moura, Leandro Pereira / da Silva, Adelino Sanchez Ramos / Ropelle, Eduardo Rochete / Cintra, Dennys Esper / Pauli, José Rodrigo

Cellular and molecular life sciences : CMLS

2023 Volume 80, Issue 5, Page(s) 122

Abstract: Objective: Intriguingly, hyperinsulinemia, and hyperglycemia can predispose insulin resistance, obesity, and type 2 diabetes, leading to metabolic disturbances. Conversely, physical exercise stimulates skeletal muscle glucose uptake, improving whole- ... ...

Abstract	Objective: Intriguingly, hyperinsulinemia, and hyperglycemia can predispose insulin resistance, obesity, and type 2 diabetes, leading to metabolic disturbances. Conversely, physical exercise stimulates skeletal muscle glucose uptake, improving whole-body glucose homeostasis. Therefore, we investigated the impact of short-term physical activity in a mouse model (Slc2a4 Methods: Slc2a4 Results: When Slc2a4 Conclusions: Both short-term exercise protocols were efficient in whole-body glucose homeostasis and insulin resistance. While endurance exercise plays an important role in transcriptome and mitochondrial activity, strength exercise mostly affects post-translational mechanisms and protein synthesis in skeletal muscle. Thus, the performance of both types of physical exercise proved to be a very effective way to mitigate the impacts of hyperglycemia and hyperinsulinemia in the Slc2a4
MeSH term(s)	Mice ; Animals ; Insulin Resistance ; Diabetes Mellitus, Type 2/genetics ; Diabetes Mellitus, Type 2/metabolism ; Muscle, Skeletal/metabolism ; Hyperglycemia/genetics ; Hyperglycemia/metabolism ; Glucose/metabolism ; Glucose Transporter Type 4/metabolism
Chemical Substances	Glucose (IY9XDZ35W2) ; Slc2a4 protein, mouse ; Glucose Transporter Type 4
Language	English
Publishing date	2023-04-13
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	1358415-7
ISSN	1420-9071 ; 1420-682X
ISSN (online)	1420-9071
ISSN	1420-682X
DOI	10.1007/s00018-023-04771-2
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Article ; Online: Antioxidant and Neuroprotective Effects of the First Tryptophyllin Found in Snake Venom (

Dematei, Anderson / Costa, Samuel Ribeiro / Moreira, Daniel C / Barbosa, Eder Alves / Friaça Albuquerque, Lucas F / Vasconcelos, Andreanne G / Nascimento, Tiago / Silva, Pedro Costa / Silva-Carvalho, Amandda É / Saldanha-Araújo, Felipe / Silva Mancini, Mariana Camargo / Saboia Ponte, Luis Gustavo / Neves Bezerra, Rosangela Maria / Simabuco, Fernando Moreira / Batagin-Neto, Augusto / Brand, Guilherme / Borges, Tatiana Karla S / Eaton, Peter / Leite, José Roberto S A

Journal of natural products

2022

Abstract: In this study, we report the isolation, characterization, and synthesis of the peptide BmT-2 belonging to the tryptophyllins family, isolated from the venom of the ... ...

Abstract	In this study, we report the isolation, characterization, and synthesis of the peptide BmT-2 belonging to the tryptophyllins family, isolated from the venom of the snake
Language	English
Publishing date	2022-12-12
Publishing country	United States
Document type	Journal Article
ZDB-ID	304325-3
ISSN	1520-6025 ; 0163-3864
ISSN (online)	1520-6025
ISSN	0163-3864
DOI	10.1021/acs.jnatprod.2c00304
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Article ; Online: Short-term strength exercise reduces the macrophage M1/M2 ratio in white adipose tissue of obese animals.

da Costa Fernandes, Célio Junior / da Cruz Rodrigues, Kellen Cristina / de Melo, Diego Gomes / de Campos, Thais Dantis Pereira / Dos Santos Canciglieri, Raphael / Simabuco, Fernando Moreira / da Silva, Adelino Sanchez Ramos / Cintra, Dennys Esper / Ropelle, Eduardo Rochete / Pauli, José Rodrigo / de Moura, Leandro Pereira

Life sciences

2023 Volume 329, Page(s) 121916

Abstract: Obesity can exacerbate the systemic inflammatory process, leading to increased infiltration of monocytes in white adipose tissue (WAT) and polarization of these cells into pro-inflammatory M1 macrophages, while reducing the population of anti- ... ...

Abstract	Obesity can exacerbate the systemic inflammatory process, leading to increased infiltration of monocytes in white adipose tissue (WAT) and polarization of these cells into pro-inflammatory M1 macrophages, while reducing the population of anti-inflammatory M2 macrophages. Aerobic exercise has been shown to be effective in reducing the pro-inflammatory profile. However, the impact of strength training and the duration of training on macrophage polarization in the WAT of obese individuals have not been widely studied. Therefore, our aim was to investigate the effects of resistance exercise on macrophage infiltration and polarization in the epididymal and subcutaneous adipose tissue of obese mice. We compared the following groups: Control (CT), Obese (OB), Obese 7-day strength training (STO7d), and Obese 15-day strength training (STO15d). Macrophage populations were evaluated by flow cytometry: total macrophages (F4/80+), M1 (CD11c), and M2 (CD206) macrophages. Our results demonstrated that both training protocols improved peripheral insulin sensitivity by increasing AKT phosphorylation (Ser473). Specifically, the 7-day training regimen reduced total macrophage infiltration and M2 macrophage levels without altering M1 levels. In the STO15d group, significant differences were observed in total macrophage levels, M1 macrophages, and the M1/M2 ratio compared to the OB group. In the epididymal tissue, a reduction in the M1/M2 ratio was observed in the STO7d group. Overall, our data demonstrate that 15 days of strength exercise can reduce the M1/M2 ratio of macrophages in white adipose tissue.
MeSH term(s)	Mice ; Animals ; Adipose Tissue ; Inflammation ; Adipose Tissue, White ; Obesity/therapy ; Macrophages ; Insulin Resistance ; Mice, Inbred C57BL ; Mice, Obese
Language	English
Publishing date	2023-07-05
Publishing country	Netherlands
Document type	Journal Article
ZDB-ID	3378-9
ISSN	1879-0631 ; 0024-3205
ISSN (online)	1879-0631
ISSN	0024-3205
DOI	10.1016/j.lfs.2023.121916
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Article ; Online: Activation of the α7 Nicotinic Acetylcholine Receptor Prevents against Microglial-Induced Inflammation and Insulin Resistance in Hypothalamic Neuronal Cells.

Amaral, Camila Libardi do / Martins, Ísis de Cássia Alves / Veras, Alana Carolina Costa / Simabuco, Fernando Moreira / Ross, Michael Glenn / Desai, Mina / Ignácio-Souza, Leticia Martins / Milanski, Marciane / Torsoni, Adriana Souza / Torsoni, Marcio Alberto

Cells

2022 Volume 11, Issue 14

Abstract: Neuronal hypothalamic insulin resistance is implicated in energy balance dysregulation and contributes to the pathogenesis of several neurodegenerative diseases. Its development has been intimately associated with a neuroinflammatory process mainly ... ...

Abstract	Neuronal hypothalamic insulin resistance is implicated in energy balance dysregulation and contributes to the pathogenesis of several neurodegenerative diseases. Its development has been intimately associated with a neuroinflammatory process mainly orchestrated by activated microglial cells. In this regard, our study aimed to investigate a target that is highly expressed in the hypothalamus and involved in the regulation of the inflammatory process, but still poorly investigated within the context of neuronal insulin resistance: the α7 nicotinic acetylcholine receptor (α7nAchR). Herein, we show that mHypoA-2/29 neurons exposed to pro-inflammatory microglial conditioned medium (MCM) showed higher expression of the pro-inflammatory cytokines IL-6, IL-1β, and TNF-α, in addition to developing insulin resistance. Activation of α7nAchR with the selective agonist PNU-282987 prevented microglial-induced inflammation by inhibiting NF-κB nuclear translocation and increasing IL-10 and tristetraprolin (TTP) gene expression. The anti-inflammatory role of α7nAchR was also accompanied by an improvement in insulin sensitivity and lower activation of neurodegeneration-related markers, such as GSK3 and tau. In conclusion, we show that activation of α7nAchR anti-inflammatory signaling in hypothalamic neurons exerts neuroprotective effects and prevents the development of insulin resistance induced by pro-inflammatory mediators secreted by microglial cells.
MeSH term(s)	Animals ; Benzamides ; Bridged Bicyclo Compounds ; Glycogen Synthase Kinase 3/metabolism ; Hypothalamus/metabolism ; Inflammation/pathology ; Insulin Resistance ; Mice ; Microglia/metabolism ; Neurons/metabolism ; alpha7 Nicotinic Acetylcholine Receptor/metabolism
Chemical Substances	Benzamides ; Bridged Bicyclo Compounds ; PNU-282987 ; alpha7 Nicotinic Acetylcholine Receptor ; Glycogen Synthase Kinase 3 (EC 2.7.11.26)
Language	English
Publishing date	2022-07-14
Publishing country	Switzerland
Document type	Journal Article ; Research Support, Non-U.S. Gov't ; Research Support, N.I.H., Extramural
ZDB-ID	2661518-6
ISSN	2073-4409 ; 2073-4409
ISSN (online)	2073-4409
ISSN	2073-4409
DOI	10.3390/cells11142195
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