Effects of PTEN inhibition over neuroinflammation in glial cells.

Author(s)

Amanda Galvão da Paixão

• Document type: Master's Dissertation
• Press: São Paulo.
• Institution: Universidade de São Paulo (USP). Instituto de Ciências Biomédicas (ICB/SDI)
• Defense date: 2018-11-09

Examining board members

Elisa Mitiko Kawamoto Iwashe; Pedro Augusto Carlos Magno Fernandes; William Tadeu Lara Festuccia; Tatiana Rosado Rosenstock

• Advisor: Elisa Mitiko Kawamoto Iwashe

Abstract

Formerly exclusively proposed as cytoplasmic phosphatase, it is now clear that PTEN function is beyond its phosphatase activity and it may be cell compartment-dependent. Growing evidences point to the modulatory potential of PTEN over inflammation. This study aimed to investigate the modulatory effects of PTEN over neuroinflammation and the role of its subcellular compartmentalization (nucleus/cytoplasm) in glial cells. Primary rat glial cells received BpV(pic) and LPS co-treatment for periods of 4h and 24h. U87MG cells (PTENnull) were challenged with LPS and transfected with plasmids for PTEN subcellular localization. The effects were analyzed by cellular viability, Western Blotting, immunofluorescence, qPCR, ELISA and Multiplex assays. The inhibition eficiency of PTEN by BpV(pic) was confirmed by increased AKT phosphorylation. Twenty-four hours post co-treatment, a reduced glial response to LPS was observed to IL-1<font face = \"symbol\">b and TNF-<font face = \"symbol\">a cytokines both at mRNA and protein levels as well as induction of enzyme Arginase-1 and attenuated expression of CD206 receptor. In addition, at the time of 24h, an induction of the antioxidant enzymes SOD2 e GSR was observed in co-treated cells. At 4h of co- treatment, PTEN inhibition alone induces IL-6 secretion. Both BpV(pic) alone and LPS co-treatment did not interfere on IL-10 signalization and in NO levels neither. Interestingly, LPS challenged U87MG (PTENnull) cells did not respond by p65 translocation or TNF- <font face = \"symbol\">a secretion, but it reduced cellular viability in high doses (100 <font face = \"symbol\">mg/mL). Cell transfection with plasmids for PTEN subcellular localization suggests that U87MG lineage are not easily transfectable by protocols tested in this study. Pharmacologic PTEN inhibition proved to be an interesting target to LPS glial response attenuation. Complementary analysis by other BpV(pic)-treatment regimens and using a model of PTENnull cells more easily transfectable may take to a better understading of the data we got so far. (AU)

• FAPESP's process: 16/07376-4 - Evaluation of the effects of PTEN compartmentalization in human cells of glioblastoma (U87MG) submitted to a inflammatory stimuli
• Grantee: Amanda Galvão da Paixão
• Support Opportunities: Scholarships in Brazil - Master