The main interest of the laboratory is to study the control of gene expression in the parasite Leishmania employing global comparative approaches, which evaluate the genome expressed under various conditions and reverse genetics. Website: https://parasitomol.fmrp.usp.br/

We investigate processes and factors involved in the gene expression control in the pathogenic protozoan Leishmania and employ large-scale approaches and reverse genetics to understand parasite biology and its interaction with the host. From the analysis of transcriptomes at different stages of parasite development, we identify computationally differentially expressed transcripts and, among them, putative non-coding RNAs (ncRNAs). We are interested in functionally evaluating these differentially expressed ncRNAs, understanding the biogenesis of these transcripts and their possible involvement in the regulation of the expression of other genes. Moreover, we intend to understand the role of enzymes that act indirectly modulating gene expression. These proteins are arginine methyltransferases (PRMTs) that transfer a methyl group to arginine residues in specific domains (rich in arginine and glycine), modifying mainly histones and RNA binding proteins involved in the regulation of gene expression. A third line recently started in the laboratory investigates the potential non-canonical activity of ribosomal proteins.

The Laboratory was inaugurated in 2006 when Dario S. Zamboni was hired by the Ribeirão Preto Medical School of the University of São Paulo (FMRP/USP). We investigate the interactions between intracellular pathogens with host cells. It is an area of multiple possibilities among the disciplines of Immunology, Cell Biology, and Microbiology. The pathogens investigated include intracellular parasites (Leishmania and Trypanosoma cruzi), intracellular bacteria (Legionella and Coxiella), and some viruses such as Mayaro and SARS-CoV-2. Researchers linked to the laboratory investigate how the immune system detects microbial pathogens and operates to eliminate the infection. We also investigate how intracellular pathogens subvert host cell roles to replicate within cells, a process that often culminates in disease onset. To achieve these goals, we use modern tools of cellular and molecular biology, biochemistry, and genetics. The laboratory is located at the Department of Cellular and Molecular Biology, FMRP/USP, and is funded by FAPESP, CNPq, CAPES, WHO, FAEPA, and PEW. The laboratory is part of the Center for Research in Inflammatory Diseases (CRID/FAPESP) and the National Institute of Science and Technology in Vaccines (INCTV/CNPq). Webpage: http://lpm.fmrp.usp.br/

ROLE OF MYOSIN-VA AND ITS LIGHT CHAIN DLC2 IN CELL MIGRATION AND APOPTOSIS FUNCTIONS

One of the current goals of our laboratory is to define the role of V-myosins in the cascade of invasion and metastasis in melanoma and carcinomas. We are characterizing the effect of siRNAs against myosin-Va and its DLC2 light chain in tumor cell lineages, as well as the mechanisms of action, properties, and anti-tumor efficacy of an apoptosis-inducing peptide derived from myosin-Va. We employ multiple approaches, including recombinant DNA construction, mammalian cell lentiviral transfection and transduction, RNAi silencing, confocal fluorescence microscopy and electron microscopy, polyclonal antibody production, and protein-protein interaction assays. IDENTIFICATION AND CHARACTERIZATION OF NEW GENES INVOLVED IN MELANOMA PROGRESSION. Tumor progression is a process characterized by the acquisition of multiple genetic changes. Unveiling these changes is a necessary step to identify new therapeutic targets and develop targeted therapeutic strategies for greater effectiveness and specificity in the fight against cancer. Our projects aim to identify and functionally characterize new genes involved in tumorigenesis and metastasis, using, as an experimental model, melanoma lineages of different stages of tumor progression, in a multidisciplinary approach, which combines methodologies of Molecular Biology, Bioinformatics, Biochemistry, and Cell Biology. GRISCELLI SYNDROME: MOLECULAR DIAGNOSIS AND FUNCTIONAL STUDIES. Three subtypes of hereditary diseases are included under the name Griscelli syndrome, GS1, GS2, and GS3, involving the genes MYO5A, RAB27A, and MLPH. They are diseases marked by pigment alteration that can occur in association with neurological deficiency (GS1), fatal immunodeficiency (GS2), or in isolation (GS3). Defects in the release of lytic granules in T-cytotoxic and natural killer cells are the basis of immunodeficiency associated with Griscelli’s Syndrome and other primary hereditary immunodeficiencies, which have as their main manifestation, the hemophagocytic syndrome, a threatening condition of sudden onset and fatal outcome if untreated, with the possibility of cure only by bone marrow transplantation. Our work has contributed to clinical areas for cellular and molecular diagnosis, determining cellular changes and mutations associated with Griscelli Syndrome. Our proposal includes extending the investigation to other hemophagocytic syndromes and the development of strategies for treatment, based on gene therapy (demonstrated at the cellular level in cytotoxic T lymphocytes in the work of our group) or protein therapy. We are also interested in studying the molecular mechanisms involved in cytoskeleton polarization and exocytosis of lytic granules in cytotoxic response.

The main areas of activity of the laboratory are pathogenesis studies, cellular and molecular biology of rhinovirus, enterovirus (Coxsackie B5 virus) and Oropouche virus, molecular biology of picornavirus, the pathogenesis of respiratory syncytial virus, and production of recombinant inputs to evaluate the immune response against Oropouche virus and enterovirus. Recently, we developed research on the biology of Leishmania viruses.

1) Patogênese e persistência de vírus respiratórios humanos Focamos na patogênese de vírus respiratórios humanos, inclusive dos vírus pandêmicos influenza e coronavírus, bem como em mecanismos de sua persistência em tecidos linfo- hematopoiéticos. Usamos análises teciduais, métodos moleculares, bioquímicos e de virologia clássica, aplicados a modelos de infecção in vitro, in vivo e ex vivo. São usadas também análises de (meta)genomas e transcriptomas em single cells. 2) Patogênese do vírus Oropouche Pesquisamos in vitro e in vivo a relação vírus-célula e a patogênese de infecções pelo arbovírus Oropouche. Nesta linha de pesquisa utilizamos infecções in vitro para investigar funções de proteínas virais não-estruturais, genética reversa, técnicas bioquímicas e de microscopia para investigar apoptose, bem como modelos animais para investigar a genética de atenuação do vírus. 3) Transmissão vertical de vírus Investigamos a transmissão vertical dos arbovírus Chikungunya e Oropouche, bem como dos vírus respiratórios influenza e sincicial respiratório. Usamos modelos experimentais em camundongos, além de testagem de amostras humanas e infecções em culturas primárias de células humanas. 4) Relação vírus-hospedeiro de vírus de Leishmania Pesquisamos os efeitos celulares de vírus que persistem no parasita Leishmania e como esta infecção altera a entrada e a replicação do parasita com macrófagos in vitro. 1) Pathogenesis and persistence of human respiratory viruses We focus on the pathogenesis of human respiratory viruses, including pandemic viruses, influenza, and coronavirus, as well as mechanisms of their persistence in lymphoid and hematopoietic tissues. We use tissue analysis, molecular, biochemical, and virology methods in vitro, in vivo, and ex vivo infection models. We also use analyses of (meta)genomes and transcriptomes in single cells. 2) Oropouche pathogenesis We investigate in vivo and in vitro the virus-cell relationship and the pathogenesis of infections by the arbovirus Oropouche. In this line of research, we use in vitro infections to investigate functions of non-structural viral proteins, reverse genetics, biochemical, and microscopy techniques to investigate apoptosis as well as animal models to investigate the attenuation genetics of the virus. 3) Vertical transmission of viruses We investigate the vertical transmission of Chikungunya and Oropouche arboviruses, as well as respiratory influenza and respiratory syncytial viruses. We use experimental models in mice, as well as testing human samples and infections in primary cultures of human cells. 4) Leishmania virus-host relationship We investigate the cellular effects of viruses that persist in the Leishmania parasite and how this infection alters the entry and replication of the parasite with macrophages in vitro.

He has a Bachelor’s degree in Chemistry (2005), a Bachelor’s degree in Biotechnology (2009), and a postgraduate degree in Biological Sciences (2014) from the Facultad de Ciencias Bioquímicas y Farmacéuticas of the National University of Rosario, Argentina. . He worked as a postdoctoral fellow at the Chemistry Institute of the University of São Paulo (2014-2019) under the guidance of Prof. Shaker Chuck Farah. He worked as Professor of the course ‘A proposal to read, write, and learn better when entering the University’ (2008-2010) and as Assistant at the Department of Organic Chemistry (2010-2012) at the Faculty of Biochemical and Pharmaceutical Sciences of the University National of Rosario, Argentina. He also served as Assistant to the Chair of Cellular Biology and Bromatology Laboratory (2012-2013) of the University Technician in Chemistry course at the Instituto Politécnico Superior ‘General San Martin’, dependent on the National University of Rosario, Argentina. He completed an internship (2016-2017) at the Department of Biological Sciences at Birkbeck College in London, United Kingdom, where he held the position of Associate Research Fellow. He currently works as a Professor at the Department of BioMolecular Sciences at the Faculty of Pharmaceutical Sciences of Ribeirão Preto at the University of São Paulo. He has experience in the areas of Molecular Biology and Structural Biology, Biochemistry and Genetics of microorganisms, focusing on complexes associated with membranes and the processes in which they are involved.

Molecular Evolution- Development Genes- Evolution and Development (Evo-Devo)

Our laboratory studies mechanisms of signal transduction and regulation of gene expression in bacteria and how these pathways regulate the physiology and virulence of the opportunistic pathogen Chromobacterium violaceum.

The study of signal transduction pathways and transcription factors has a major impact on understanding how bacteria control various cellular functions such as biofilm production, virulence, antibiotic resistance, and host recognition. More specifically, we use Chromobacterium violaceum, a free-living beta-proteobacteria capable of acting as an opportunistic pathogen in humans, as a model to study the role of transcription factors in virulence. We employ several experimental approaches, such as transposon mutant library scanning, specific mutagenesis, DNA microarrays, and virulence assays in animal models to understand the various aspects of the biology of this organism, including its relationship with the host. Currently, the main lines of research include (I) identification of transcription factors, with a focus on redox regulatory that affect the virulence of C. violaceum and identification of their regulons; (ii) determination of the role of eukaryotic-type kinases/phosphatases (Ser/THR/Tyr kinases) in the regulation of the activity of these transcription factors to understand whether they can integrate different signals (redox and phosphorylation regulation) in virulence control.

The aim of Dr. Sales’ Laboratory is to understand the cellular and genetic mechanisms involved in cancer initiation in the mouth and uterine cervix, with special focus on serine protease and its inhibitors.

The high incidence of epidermoid carcinoma, its infiltrative character, possibility of causing metastases and low survival justify the investment in understanding the proteolytic pathways involved in this neoplastic process. Serine proteases and their inhibitors, crucial for the maintenance of healthy tissues, act as pivots of tissue destruction in many pathologies, which include malignant neoplasms. In this sense, serine proteases are useful as prognostic markers in different types of cancers, where the proteolytic activity of these enzymes is unbalanced. Through translational research using animal models (transgenic mice), cell and tissue biology, we search for information to support the design of appropriate clinical interventions and the development of resolution therapies.

We study molecular and cellular processes involved in the differentiation of queens and workers in social bees to understand plasticity issues in development.

Honey bees have attracted the attention of several research groups not only for their economic and ecological importance (pollination) but also for having a high potential for genetic studies. Due to its social organization, broad behavioral plasticity, a large number of individuals by colonies, and diversity of molecular techniques, it is possible to study the regulation of gene expression of genes involved in metabolic cascades important for the maintenance of physiological homeostasis and post-embryonic development, and also analyze other molecular mechanisms absent in model organisms, such as DNA methylation, which provide evidence that helps to understand the molecular processes in the social organization of these insects. Our goal is to research processes involved in the control of the differential development of honey bee varieties. Currently, the main focus of our research is on the regulation of gene expression from the differential feeding that occurs between the larvae of queens and workers. The complete sequencing of the Apis mellifera genome allowed functional genomic studies on social organization, phenotypic plasticity, and longevity. Through gene expression analysis with emphasis on oxidative metabolism, epigenetics, and ovarian development, we aim to elucidate key processes of larval development and metamorphosis, describing gene networks that control the development of target tissues. Phenotypic plasticity and the division of labor among adult workers are directly linked to the synthesis of vitellogenin, juvenile hormone titers, and the insulin/TOR signaling module. Using approaches such as RNAi, transcriptome, real-time PCR, Western blot, and microscopy, we aim to understand the interaction between these controlling components of the bee life cycle.

Investigation of extra and intracellular signalling mechanisms and their role in the regulation of the innate immunity activation.

In our lab, we use a multidisciplinary approach working towards understanding how signaling transduction cascades initiate, progress and ultimately regulate the function of innate immune cells upon activation of a primary function such as phagocytosis. This knowledge is used to gain insight into the pathogenesis of inflammatory disorders (such as autoimmune diseases and cancer), as well as in the mechanisms driving the balance between resistance and tolerance to infections. We currently aim to use cellular and molecular biology, protein biochemistry and immunoassays to investigate the role of the autophagy machinery, a bonafide mechanism of coping with metabolic stresses, in the regulation of phagocytosis by innate immune cells. Phagocytosis of invading microorganisms and dead cells is a primordial function of the immune system, and the phagosome has emerged as an autonomous signaling compartment within the cells. During phagocytosis, diverse components of the autophagy machinery can be recruited to the phagosome membrane, ultimately impacting cell function. In this context, our main current research projects include:1. To determine the mechanisms of LC3-associated phagocytosis (LAP) induction and LAPosome maturation.2. To investigate the molecular mechanisms for the regulation of macrophage function by LAP.3. To investigate the consequences of LAP activation in innate immune cells in the pathogenesis of inflammatory diseases and cancer.

The focus of our laboratory is to functionally characterize proteins that may be involved in leukemogenesis. Our main challenge at the moment is to implement the methodology of bone marrow transplantation in a murine model. This model is an excellent tool to study leukemogenesis since it mimics in vivo the neoplastic transformation that occurs in the bone marrow in humans, and thus allows to test the transforming ability of several proteins in the hematopoietic system. The technique is based on the use of retrovirus for ex vivo gene transfer in hematopoietic progenitor cells and its inoculation in irradiated mice with sublethal doses. The transplanted cells reconstitute the bone marrow and hematopoiesis of the mouse and depending on the transformative potential of the oncogene or the gene lesion introduced into the progenitor cells, the development of malignant clones and triggering of the leukemic process in the animal will occur. Webpage: https://sites.usp.br/limca/

1-Investigation of the participation of the regulatory Splicing Factor Kinases (KIS; UHMK1) in leukemogenesis: Changes in the splicing process constitute a new and important mechanism in leukemogenesis. Preliminary data from our group suggest the participation of KIS in the differentiation of hematopoietic cells. We hypothesize that through the phosphorylation of splicing factors, KIS affects the splicing of genes important for the differentiation of hematopoietic cells. To evaluate this issue we will use a murine model of bone marrow transplant and in vitro assays. 2-Characterization of the function of splicing factors in the normal and abnormal hematopoietic system: The splicing factor SF1 is a fundamental component in the assembly of the prespliceosome, mutated in some patients with hematological neoplasms. Without function described in the hematopoietic system, our group aims to evaluate the functional consequence of inhibition and overexpression of SF1, as well as its mutated variants in leukemic cell line models. 3-Investigation of cooperative oncogenes to IL7R mutations in acute leukemias. Mutations in the interleukin-7 receptor (IL7R) gene result in the constitutive activation of this receptor and its signaling pathway. Our group is interested in identifying genes that cooperate with mutations in IL7R to accentuate or inhibit the malignant phenotype. For this, Ba/F3 lineages carrying wild and mutated IL7R will be transduced with lentiviral particles expressing an shRNA library. Cells that exhibit accelerated or decreased growth will be sequenced to identify oncogenic partners or IL7R oncogene suppressors, respectively. Additionally, mutated IL7R will be introduced in conjunction with other genetic lesions of interest in hematopoietic progenitor cells, which will be used in a murine bone marrow transplant model and in vitro assays.

The laboratory studies the molecular mechanisms involved in addressing proteins to intracellular compartments of the secretory and endocytic pathways, and the subversion of these processes by viruses.

We are interested in understanding molecular mechanisms involved in addressing membrane proteins to intracellular compartments of the biosynthetic/secretory and endocytic pathways in different cell types. These processes have a fundamental role in the biogenesis and maintenance of the functioning of organelles that make up the endomembrane system. Changes in load recognition in these intracellular transport pathways are implicated in various genetic, degenerative diseases, and also in virus infections. Our current work is focused on the following topics: 1) Identification of mechanisms involved in the negative regulation of cell surface proteins by HIV-1 accessory proteins Nef and Vpu. 2) Identification of cellular factors and targeting signs involved in intracellular glycoprotein traffic of the HIV envelope. 3) Elucidation of the mechanisms involved in the assembly and budding of disease-causing Bunyaviruses.

Our laboratory studies the genomic plasticity and its relationship with response mechanisms to DNA damage in the protozoan Leishmania.

The line of research developed in our laboratory aims at understanding the pathways of detection and signaling of damage in the DNA of the protozoan parasite Leishmania. We hypothesize that the peculiarities of these pathways are determinants of the plasticity of the genome of this organism. More specifically, we investigated the parasite proteins that could compose a homologous complex to the 9-1-1 clamp, found in higher eukaryotes. In mammalian and yeast cells, the 9-1-1 clamp is central to the response to replicative stress because it signals the shutdown of the cell cycle and participates in the recruitment of DNA repair machinery. We found that the proteins LmHus1 and LmRad9 may compose a possible 9-1-1 homolog and participate in the response to DNA damage.

Biotechnology plays a crucial role in the development of biocatalysts or genes that can expand the limits of life for use in industry, agriculture, medicine, and power generation. Currently, there has been a growing demand for enzymes and microorganisms with better catalytic performance or tolerance to specific parameters of industrial processes. Metagenomics takes advantage of the richness of the genetic and biochemical diversity present in the genomes of microorganisms found in nature and provides a set of new technologies aimed at screening new catalytic activities with potential biotechnological applications. However, the biased and low level of heterologous protein expression in Escherichia coli, along with the use of non-optimal metagenomic screening strategies, often results in a low success rate in identifying new enzymes or other genes of industrial interest. In this sense, our research group aims to develop new tools to improve the recovery of target genes. Fundamentally, we carry out gene prospecting with biotechnological potential by screening metagenomic libraries using synthetic biology approaches.

1. Novel approaches to improve functional screening of biocatalysts in metagenomic libraries

2. Screening of new regulatory elements for use in Synthetic Biology

3. Screening of new genes expanding the limits of life in bacteria

4. Engineering yeast strains to improve stress-tolerance

5. Systems microbiology approaches for mapping the antibiotic resistance in Brazil

Studies on genetic-molecular changes of malignant neoplasms, and search for potential therapeutic targets for their treatment. This line of research investigates the in vitro and in vivo effects of inhibition of different oncoproteins (such as NF-kB, PLK1, AP-1, ROCKs, etc.), and microRNAs that may assist in the development of alternative therapies for cancer.

The approaches are multiple involving treatments with ionizing radiation and innovative antitumor compounds focusing on cellular responses at the level of cell cycle control, mechanisms of death, changes in gene expression, clastogenicity, and the evaluation of other cellular parameters such as viability, clonogenic, and invasive capacity of tumor cells.

Investigation of RANK-RANKL signaling as an endocrine mediator of bone in different organs, especially in mitochondrial biogenesis and its impact on the differentiation of beige adipocytes, immunometabolism with focus on the polarization of macrophages between pro and anti-inflammatory states, and the differentiation of skeletal striated muscle.

The RANKL system consists of a triad of proteins composed of two receptors: RANK and Osteoprotegerin (OPG), and a ligand: RANKL. This system is commonly related to bone metabolism; however, it is present in several tissues and regulates communication between immune cells, development of mammary glands, and, recently, we have demonstrated its role in calcification of arteries, and prevention of neuronal death after cerebral ischemia. OPG has been considered a biomarker in obesity, type 2 diabetes, cardiovascular diseases, and cerebral ischemia, but the biological mechanism that explains the increase in the level of OPG in these different pathologies remains unclear. Our aim is to evaluate the RANK-RANKL pathway in macrophages, adipose tissue, and skeletal muscle in the physiological context and insulin resistance. 1) Phenotypic change of macrophages in adipose tissue inflammation: The adipose tissue of obese individuals has a large infiltrate of pro-inflammatory macrophages (M1), which contribute to insulin resistance in this tissue. Our goal is to elucidate the molecular mechanism of RANKL signaling responsible for interfering with the TLR4 pathway and inhibiting the M1 profile and evaluate its contribution in adipose tissue inflammation in murine models of diet-induced type 2 diabetes. 2) Differentiation of beige adipose tissue and its signaling pathways. The differentiation from white adipocyte to beige adipocyte in the process known as browning is considered a therapeutic target to treat obesity and related diseases since beige adipocytes have a low accumulation of lipid droplets, thermogenic capacity, and increased energy expenditure since they dissipate energy from cellular respiration in the form of heat. We investigate the adipogenesis and differentiation of white and beige adipocytes, and mitochondrial biogenesis pathways regulated by RANKL and elucidate possible interaction between bone and adipose tissue. 3) Differentiation and determination of the type of skeletal striated muscle fiber. We investigate the biogenesis and mitochondrial degradation pathways to determine the type of skeletal striated muscle fiber in OPG knockout or heterozygous mice to understand the molecular mechanism of interaction between bone and muscle metabolism, and the influence of the RANK-RANKL axis on insulin response in these cells, since the problem in glucose uptake by skeletal muscle, in addition to adipose tissue, is a target for the treatment of Type 2 diabetes.

The laboratory focuses on the functional characterization of cytoskeleton proteins of relevant pathogens in our environment such as Leishmania, Trypanosoma cruzi, and Trypanosoma brucei and in cellular and molecular interactions with the host cell.

1) Project characterization of cell compartments during infection by Trypanosoma cruzi. Trypanosomatids have developed a range of strategies to subvert and circumvent the innate host immunity and we are interested in characterizing the cellular and molecular changes that the host cell responds to due to the presence of the parasite in the cytoplasm and nucleus. We employ numerous cell biology techniques such as fluorescence and confocal microscopy in fixed and living cells using cell compartment markers, western blotting, RNAseq, and image analysis in specific software. Also, recombinant protein construction and silencing by RNAi and gene editing by Crispr/Cas9 for cell interaction studies. 2) Project: characterize new proteins and cytoskeleton genes of relevant pathogens in our environment as Leishmania major, Trypanosoma cruzi, and Trypanosoma brucei. Trypanosomatids represent a magnificent model for analysis of basic issues about the cytoskeleton, allowing us to obtain better knowledge about the evolution of higher eukaryotes. We recently characterized the FAZ10 protein in T. brucei. This protein has a role in the positioning of the cleavage furrow during cell division and its absence leads to errors generating multinucleated or nuclei-free named zoids during the cell cycle (Moreira et al., 2017).  We are interested in identifying the molecular partners that interact with FAZ10 in the cytoskeleton. Related projects involve polyclonal antibody production, immunofluorescence, confocal, electron microscopy, western blotting, cloning, recombinant protein constructions, transfections, interference RNA (RNAi), gene editing through Crispr/Cas9, protein-protein interaction, bioinformatic analysis, among others. Contact us to learn more about the projects in the lab.

Genomic studies in Diptera. Gene expression and amplification regulated in development. Regulatory mechanisms of transcription in higher eukaryotes, with emphasis on the regulation by steroid hormones. Characterization of the expression pattern of Hox genes in the early development of Bradysia hygida. Bioprospection of glycosyl hydrolases in the dipter B. hygida.

In our laboratory, we use as a model the basal Dipterus Bradysia hygida, a sciarid isolated at the Ribeirão Preto campus in 1965, and that has been continuously kept in culture in the laboratory. In Diptera phylogeny, the infraorder Biobionomorpha, which includes the sciarids, occupies an intermediate position between the infraorders Culicomorpha (including the mosquitoes vectors of diseases) and Brachycera (including the genera Musca and Drosophila), which makes the study of sciarids relevant in the context of Diptera evolution. Sciarids are used as models to investigate molecular mechanisms that occur only in the family Sciaridae (gene amplification in regions forming DNA puffs, elimination of chromosomes in the process of sexual determination), as well as to characterize processes in Metazoa (programmed cell death, nucleolar organization, telomere structure, among others). The family Sciaridae comprises about 20,000 species in which only 2,000 have been described, and 201 species distributed in 29 genera have been described in Brazil. The most recent phylogeny of the family Sciaridae revealed the evolutionary history of the larval habitats of sciarids, which include everything from decaying organic matter to fungi and living plants. Also, as numerous species of sciarids cause damage to agriculture, the study of sciarids has the potential to contribute to the establishment of control strategies of sciarid species that are agricultural pests. The genome of B. hygida has been recently sequenced and is in the final annotation phase. The availability of quality genomic data, such as those obtained by our laboratory, allows us to investigate different issues in this model. Genomic data associated with salivary gland transcriptome data are beingused to characterize DNA puff-forming regions. This line of research will allow us to expand the molecular characterization of amplified domains in DNA puffs and contribute to the understanding of the mechanisms that promote the additional replication of these regions of the genome. Another line of research of the laboratory aims to characterize the transduction pathway induced by the hormone ecdysone at the entrance to metamorphosis. Unlike what occurs in different model organisms (e.g., D. melanogaster, A. mellifera, B. germanica, B. mori), where the role of ecdysone as a metamorphosis regulator has been widely characterized. In sciarids, the ecdysone also acts as a factor that promotes transcription and amplification of DNA puff genes at the end of the fourth larval stage. The genomic data are being used as a starting point for the annotation of genes that are part of the transduction pathway regulated by ecdysone and design experiments to investigate the temporal and tissue patterns of expression of these genes in B. hygida. Together, the data obtained will expand the characterization of the role played by ecdysone in the sciarid. B. hygida is also a relevant model for studies in the area of Evolutionary Developmental Biology. The description of the embryonic development, which lasts 9 days at 22 °C, which is associated with the genomic data, and the transcriptome of each of the first six days of embryonic development to be used as a starting point for the characterization of the Hox genes, which are transcription factors that determine the identity of the segments of the insect, and occupy apical positions in networks regulating gene expression, and promotes the development of a metazoan. Preliminary data suggest that the organization of Hox genes in B. hygida is distinct from that found in other Diptera. On the other hand, initial immunolocalization experiments indicate that the expression pattern of bithorax complex genes during embryonic development is conserved in the sciarid. Data from the characterization of Hox genes in B. hygida will contribute to new information about the evolution of the genomic organization and Hox expression patterns in insects, a topic that remains current and relevant in the area of Evolutionary Developmental Biology. Bioprospection is a strategy that allows the discovery of hydrolytic enzymes in environmental samples that can be used to improve efficiency and decrease costs in different processes, including the production of second-generation ethanol. In nature, B. hygida acts as a decomposer, and throughout larval development, the larvae of B. hygida feed on partially decomposed organic matter. Thus, the last line of research underway in the laboratory is aimed at bioprospection of hydrolytic enzymes in larvae of B. hygida. Biochemical assays using as substrates synthetic compounds and purified polysaccharides revealed a set of catalytic activities in the larvae of B. hygida. This characterization is being expanded to identify which larval tissues present the activities initially identified, and in a subsequent step, the data obtained will be correlated with analyses designed to identify coding sequences of hydrolytic enzymes in the genome of B. hygida. Thedevelopment of this project, therefore, has the potential to contribute to the identification and characterization of new hydrolytic enzymes capable of converting lignocellulosic material into reducing sugars more efficiently to be used in different applications.

The Computational Chemical Biology Laboratory (CCBL) applies and develops computational statistical methods to understand the chemical composition of a variety of biological systems. Site: http://ccbl.fcfrp.usp.br/ccbl/index.html

The Laboratory of Computational Chemical Biology develops research on data sciences to integrate different sources of information and use computational statistics to elucidate metabolic pathways altered under different sampling conditions in different organisms, in particular, plants and microorganisms.

Chronic pain is an important clinical condition since there are no effective treatments. Also, the drugs available on the market have a number of side effects that limit their use. In this sense, the study of the cellular and molecular mechanisms involved in the development of chronic pain can certainly point to new targets for the development of more effective drugs with fewer side effects. In this sense, our research group has been developing projects that involve the characterization of these mechanisms through approaches in vivo and in vitro with pharmacological and genetic/molecular tools and the most modern methodologies RNA-scope, single-cell RNA-seq, CRISPR/CAS9, electrophysiology, among others.

1. Neuro-immune-glial interactions in the development of pathological pain.
2. Role of cell metabolism in neuroinflammation.
3. Molecular basis of peripheral analgesia.