This project is envisioned as a model for possible dissertations and theses encompassing glioblastomes, which are the most common brain tumors, corresponding to more than 40% of all neoplasms of the central nervous system (CNS). In particular, it will focus on those of astrocytic composition, astrocytomes. The mechanisms that culminate, in structural terms, in cytological atypia, altered mitoses, necrosis in the center of the tumor mass and in the palisade organization of its cells will be addressed. In a translational perspective, the project opens the perspective to investigate the correlation of these events with the rate and time of survival. It is intended to investigate the role of tumor mass stem cells in controlling tumor development and the mechanisms that make them potentially responsible for resistance to chemotherapy and radiotherapy procedures. The project thus intends to address the characterization and control of stem cells in the glioblastoma mass, proposing in vivo tests with a xenotransplantation model to, also, in a translational perspective, define the best procedure to use the cytolysin complex Echinotoxin II with each of the two drugs for clinical or experimental use against the tumor, TMZ or VP16. The search for an alternative to search for the therapeutic efficacy of the action of this drug complex on human glioblastoma may be the center of a thesis by silencing drug reflux pumps (MDRs), whose initial trials opened the prospect of proceeding with therapy with reflux blocking may be a strategy in future clinical studies. Therefore, the current project, which emerges from BIOTRANS ' three lines of research and permeates them horizontally, addresses the potential strands proposed for dissertations and theses: (i) the structural dynamics of tumor evolution by bioimage based on nuclear magnetic resonance; (ii) the cellular and molecular mechanisms of maintaining pluripotent status and inducing cell differentiation in stem cells; (iii) the control of the proliferation and differentiation of stem cells in vitro (expansion) and in situ (neoplastic transformation); (iv) the role of MDRs through a possible intersection with the project dedicated to transporting ATPases; (v) pre-clinical studies on the use of adult cells and isogenic, allogeneic or xenogenic stem cells in cell therapies.
The project, which permeates both lines of research, namely Biomarkers and Tissue Bioengineering, proposes to address the role of ion-carrying ATPases and investigate molecular mechanisms of their regulation in two major strands of study, which present ATPases as a common thematic link: (i) malnutrition and its late impact on renal and cardiovascular function; (ii) mechanisms of interaction and repair by stem cells in injured kidney tissue. The ATPases, which will be the object of study in possible dissertations and theses will be: Ca-ATPases of plasma membrane and sarco/endoplasmic reticulum, (Na + K) ATPase, Na-ATPase, multi-drug resistant ATPases (MDRs) and FoF1-ATPsintase (ATPase mitochondrial). In these dissertations and theses, the focus will be the investigation of signaling pathways involving kinases, such as PKA, PKC, MEK / ERK1 / 2, phospho-ERK1 / 2, PI3K / Akt and the salt inducible kinase (of potential relevance in a translational perspective), as well as the mechanisms of their regulatory coupling with the ATPases object of interest in each aspect of the project. The role of calcium and sodium as second messengers and not just as ionic transported species will also be investigated in the context of the two strands of the project mentioned above. In the second of these strands, we intend to investigate the role of mesenchymal cells in co-culture with renal cells in restoring the function of ion-carrying ATPases after anoxia injury followed by reoxygenation. Differential and phosphoproteomic proteomics studies are proposed to investigate the role of mesenchymal cells and induced pluripotency (iPS) in the expression of key proteins for the recovery of trans-epithelial transport after ischemia. In vivo and in vitro models involving rats, culture of human renal cells, culture of human mesenchymal cells and subcellular fractions of membranes, mitochondria and Golgi apparatus will be used, combining biochemical, cellular and proteomic techniques. With the development of the different strands in BIOTRANS 'dissertations and theses, the project aims to contribute to understanding at a new conceptual level integrated the regulatory networks of ATPases in relevant physiological and pathological situations, to explain the genesis of injuries and to stop in a translational perspective proposing new diagnostic and therapeutic approaches.
This project, designed for the development of dissertations and theses in the context of BIOTRANS, has two main aspects that are nurtured by shared ideas, objectives and methodologies and by different ongoing research projects. In the first one, it is proposed to investigate possible strategies for the investigation of the molecular bases that allow the treatment of cartilaginous lesions through the manufacture of equivalent cartilage tissue in agarose hydrogel micro-molds from mesenchymal cells and pluripotency stem cells. induced (iPS). It is intended to study how these cells interact with micro molds in cultures in three dimensions (3D) (spheroids), associated, in this way, with a methodology of printing organs and tissues with high cell density. It is proposed that possible dissertations and theses can study the mimicry resulting from the growth and interaction of two or more different tissues and the three-dimensional contact cell-cell and cell-matrix, similarly to what occurs in vivo. Therefore, it is crucial investigating factors involved in this interaction as an initial focus of the study regarding molecular dynamics of biocartilage. It is also intended, additionally, to investigate how potential complex tissue grafts are structurally organized by the fusion of hundreds of spheroids. In the second part of the project, BIOTRANS´ dissertations and theses will be able to investigate, also in a translational perspective of revascularization, the structural dynamics and the physical and physiological characteristics of small-caliber vascular bioprostheses, given the high rates of thrombosis of synthetic grafts and the exhaustibility autologous grafts. In additional subprojects, we intend to study the characteristics of the decellularization process of small-caliber arteries of medium-sized animals (the model currently required in translational biomedicine), investigating the components of the remaining extracellular matrix and its physical, biochemical and physiological behavior. It is also proposed to investigate physiological processes of biocompatibility in vivo and to develop in vitro toxicity assessments in parallel studies, according to the ISO IEC 10993 standard. In a subsequent step and in a more advanced translational perspective, the possibility of rejection, dilation, ruptures or occlusions and strategies to minimize them, making these bioprostheses a structure comparable to a healthy native artery. The two strands of the project will involve methodologies ranging from cell culture, going through the investigation of the interactions of cells with engineered platforms and the physical and structural analysis of the investigated biomaterials. Thus, it culminates in testing of medium-sized animals.
The project, which emerges and permeates BIOTRANS´ three lines of research, has two strands that converge in the dynamics of injured tissues and in the molecular mechanisms associated with possible recruitment, signaling and tissue repair in treatments with adult, embryonic stem cells, or induced pluripotency (iPS). Thus, possible dissertations and theses might aim at studying the processes of both intercellular communication and cellular mimicry, which takes place in the microenvironment in which the presence of the stem cell and the injured tissue exists, in order to understand what are the mechanisms underlying this possible interaction. In the prospect models, it is intended to evaluate environments in which the lesion was established by infectious processes of parasitic origin (Trypanosoma cruzi, Leishmania amazonensis, Toxoplasma gondii) as representations of neglected diseases. It is intended to investigate apoptotic mimicry as a strategy for a successful infection by these agents and the molecular mechanisms underlying a possible control of this process by stem cells, with an initial focus on the study of potentially cytokines, interleukins, trophic factors and microRNAs involved and related to structural modifications. Among possible dissertations and theses to be part of this project, methodological resources will be used encompassing tools for: (i) cell culture; (ii) microscopy (conventional, confocal and transmission and scanning electronics, with modern approaches); (iii) biochemistry and (iv) molecular biology. The analyzes will allow a better understanding of the migration and differentiation of stem cells, as well as a more precise analysis of the biological and ultrastructural consequences of the behavior of these cells, such as changes in cellular organelles and possible modulation of apoptotic mimicry in tissues injured by parasites, within a potential translational perspective.