Transcriptional regulation of PKM zeta kinase and its association in the establishment of chronic pain

• Grant number: 20/16204-8
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• Start date: February 01, 2021
• End date: January 31, 2023
• Field of knowledge: Biological Sciences - Morphology - Anatomy
• Principal Investigator: Camila Squarzoni Dale
• Grantee: Carolina Purcell Goes
• Host Institution: Instituto de Ciências Biomédicas (ICB). Universidade de São Paulo (USP). São Paulo , SP, Brazil
• Associated research grant: 19/06982-6 - Characterization and development of new modulators of the TrkA and PKMzeta pathways in inflammatory and chronic pain, AP.TEM

Abstract

The long-term potentiation (LTP) is one of the main mechanisms responsible for the plasticity of the nervous system, involving the strengthening and formation of lasting synapses, and an increase in the intensity of synapses. LTP also participates in brain remodeling at the level of the anterior cingulate cortex (ACC) to establish chronic pain. In this process, Protein Kinase C M zeta (PKMz) has been implicated as having a key role in the establishment of LTP. However, the exact role of this kinase in this process is still quite controversial. The locus of the gene encoding Protein kinase C zeta (PKCz) has two promoters, with different nucleotide composition. While PKCz has a ubiquitous expression pattern, being expressed in several vertebrate tissues, PKMz is an isoform restricted to the nervous system. Little is known about the mechanisms that regulate the expression of PKMz at the transcriptional level. Investigating the mechanisms of PKMz regulation may help to understand the development of chronic pain and possible future therapies. Thus, we will verify in several models of chronic pain whether there is an alteration in the expression of this kinase, since laboratory data suggest that the regulation of PKMz expression involves epigenetic mechanisms and has already been described as having an alteration in its expression in chronic pain, in addition to depression and Alzheimer's. PKMz expression will be evaluated both in cell culture and in ACC and other brain regions of chronic pain models. For this, we will assess whether epigenetic mechanisms, such as methylation and histone modifications, are involved in the regulation of the expression of this kinase through CRISPR/Cas9 and demethylating drugs. The subsequent validation will be through the manipulation of these mechanisms and the consequent observation of the modulation of PKMz expression and its effect on chronic pain. Changes in the profile of histone marks and their relationship to methylation will also be verified. The contribution of transcription factors to PKMz expression will be examined with gene silencing tools and drugs in cell culture. Finally, we will transpose the results obtained in vitro to in vivo using a murine experimental model of chronic sciatic nerve constriction. (AU)

Scientific publications

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

PRAMIO, DIMITRIUS TANSINI; VIECELI, FELIPE MONTELEONE; VARELLA-BRANCO, ELISA; GOES, CAROLINA PURCELL; KOBAYASHI, GERSON SHIGERU; PELEGRINA, DIOGO VIEIRA DA SILVA; MORAES, BEATRIZ CAROLINE DE; EL ALLAM, AICHA; DE KUMAR, BONY; JARA, GABRIEL; et al. DNA methylation of the promoter region at the CREB1 binding site is a mechanism for the epigenetic regulation of brain-specific PKM zeta. BIOCHIMICA ET BIOPHYSICA ACTA-GENE REGULATORY MECHANISMS, v. 1866, n. 1, p. 13-pg., 2023-01-25. (19/06982-6, 15/24046-5, 20/13929-1, 20/16204-8, 15/17812-3)