Development of HaCaT keratinocyte cell line knockouts for PKM2 enzyme

• Grant number: 16/10867-0
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date until: August 01, 2016
• End date until: December 31, 2017
• Field of knowledge: Biological Sciences - Immunology - Cellular Immunology
• Principal Investigator: José Carlos Farias Alves Filho
• Grantee: Gabriel Azevedo Públio
• Host Institution: Faculdade de Medicina de Ribeirão Preto (FMRP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil
• Associated research grant: 13/08216-2 - CRID - Center for Research in Inflammatory Diseases, AP.CEPID

Abstract

Keratinocytes are the cells in the epidermis that play a central role in inflammatory responses in the skin, actively participating in the development and maintenance of certain diseases such as psoriasis. In recent years, it has been shown that under inflammatory stimuli activated cells undergo a metabolic reprogramming culminating in increased expression of genes involved in glycolytic pathway, a phenomenon known as aerobic glycolysis or Warburg effect. The HIF-1± transcription factor is essential for development of the Warburg effect, it promotes the transcription of many enzymes of aerobic glycolysis, among them Pyruvate Kinase enzyme M2 (PKM2). The PKM2 shows two structural conformations, tetrameric or dimeric, having pyruvate kinase function and of a co-transcriptional factor for HIF-1± respectively. Recent studies have shown that PKM2 can be activated by inflammatory stimuli and is important for cytokine secretion and cellular proliferation. In this sense, results of a project underway in our laboratory show that PKM2 is highly expressed in the skin of patients with psoriasis and animals with this disease. However, no studies in the literature show the participation of PKM2 in keratinocyte activation. Thus, the aim of this research project is to generate a cell line of keratinocytes (HaCaT) knockout for PKM2 enzyme, which will later be used as a tool for detailed investigations of the participation of this enzyme in keratinocytes activation and of the development and maintenance of psoriasis.

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)

VERAS, FLAVIO P.; PUBLIO, GABRIEL A.; MELO, BRUNO M.; PRADO, DOUGLAS S.; NORBIATO, THAINA; CECILIO, NERRY T.; HIROKI, CARLOS; DAMASCENO, LUIS EDUARDO A.; JUNG, REBECCA; TOLLER-KAWAHISA, JULIANA E.; et al. Pyruvate kinase M2 mediates IL-17 signaling in keratinocytes driving psoriatic skin inflammation. CELL REPORTS, v. 41, n. 13, p. 23-pg., 2022-12-27. (13/08216-2, 16/10867-0)