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Single cell analyses and non-linear optics approaches of prostate biology


The prostate is susceptible to different diseases including the prostatitis, benign prostate hyperplasia and prostate cancer. Prostate cancer will affect 12.5% of men in all ages and 50% of men at the age 85. Benign prostate hyperplasia will affect a larger number of younger men. Our interest is to study prostate physiology at the cell and molecular biology levels, to better understand this organ's and its different cell types' behavior in diseases.This project is divided in four subprojects including (a)Prostate development, focusing on the cell-cell interactions taking place at the epithelial growth tip, in particular in the relationship between the differentiation of smooth muscle cells and the settlement of p63+ cells along the epithelial structure, considering the MMPs. Aspects of the mechanics of the development will be obtained from the analysis of smooth muscle cells differentiation/contractility and the deposition of collagen fibers.(b)The organ's response to castration/orchiectomy, specially in the characterization of gene networks capable to orchestrate the tissue modification taking place in the hypoandrogenic environment at the level of single cells, focusing on the expression of collagenases (MMP-1, MMP-8 and MMP-11.(c)The inflammation affecting the gland in experimental autoimmune prostatitis and in the response to diets containing different types of fatty acids, with emphasis on the macrophages and, among them, the M1142 macrophages we have previously described; this subproject will include an analysis of the microbiota, seeking for possible association with systemic and tissue inflammatory markers and(d)The structure and function of the protein ps20, expressed by smooth muscle cells, with attention to the possible activity as serine-proteinase inhibitor and other physiological functions, such as the relaxation of smooth muscle cells. In each of these subprojects, we will set the role of different cell types and their interactions, as well as identification of molecules involved, seeking to define the molecular mechanisms involved. Beyond the epithelial and stromal subtypes, we will be engaged with the identification of immune system's cells, particularly the macrophages, performing functions in the different phenomena associated with the response to castration and in the experimental autoimmune prostatitis. Our approach to these themes will be centered in two major technologies. The first is the analysis of single cell transcriptomes (scRNAseq). The second is the use of non-linear optics-based microscopies (SHG, FLIM, CARS). Implementing scRNAseq is the major methodological challenge to this project. The second is part of our expertise. In some cases, we will associate with other laboratories to perform proteomics, metabolomics and metagenomics. We will be engaged in the training of personnel and in science diffusion, considering the obtained results/data, as well technological and therapeutical progresses in the scope of the project. (AU)

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