Understanding the role of dermcidin on melanoma survival and migration using seriniquinone as a pharmacological tool

• Grant number: 23/17281-4
• Support Opportunities: Scholarships abroad - Research Internship - Master's degree
• Start date: April 01, 2024
• End date: July 31, 2024
• Field of knowledge: Biological Sciences - Pharmacology - General Pharmacology
• Principal Investigator: Leticia Veras Costa Lotufo
• Grantee: Rodrigo dos Anjos Miguel
• Supervisor: Sharon Prince
• Host Institution: Instituto de Ciências Biomédicas (ICB). Universidade de São Paulo (USP). São Paulo , SP, Brazil
• Institution abroad: University of Cape Town (UCT), South Africa
• Associated to the scholarship: 22/15832-0 - Assessment of seriniquinone delivery to melanoma cells by poly (D,L-lactic-co-glycolic) acid (PLGA) nanoparticles, BP.MS

Abstract

Seriniquinone (SQ) and its derivatives are unique dermcidin (DCD) modulators. This protein seems to be involved in a range of cancer-related processes, including cell survival, migration, and cachexia. Despite its potential as a drug candidate, based on its promising in vitro efficacy data, little is known about the mechanism of action of SQ and the involvement of DCD in cancer biology. Moreover, SQ has poor water solubility and poly (lactic-co-glycolic acid) (PLGA) nanoparticles (NPs) (PLGA-NPs) are proving to be great allies in seriniquinone (SQ) encapsulation, improvement of solubility, dispersion in aqueous-based vehicles, prolonged release, and displaying interesting toxicity results in in vivo assays. However, little is known about the effect of nanoencapsulation on the mechanism of action of this drug candidate. Considering these features, our group aims to establish melanoma models in which DCD is either stably silenced or overexpressed and to verify whether its modulation will maintain the potent anticancer effects of SQ when encapsulated in PLGA-NPs versus free-molecule and, further, elucidate the role of DCD in the mechanism of action of SQ. Cells will be silenced by transfection with a plasmid coding for DCD-specific short hairpin RNA (shRNA), which will be transcribed, processed and target DCD mRNA leading to the knockdown of its expression. DCD overexpression will be achieved by transfection with a plasmid containing the DCD coding sequence tagged with a FLAG sequence that will be useful as an epitope for tracking the DCD protein. The generation of DCD-knockdown or -overexpressed stable cell lines will be confirmed by evaluating DCD mRNA expression by reverse transcriptase quantitative polymerase chain reaction (RT-qPCR) and by assessing DCD protein expression by Western blotting.