Validation of molecular targets involved in inflammation and nociception using CRISPR/CAS9

• Grant number: 25/21171-5
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• Start date: November 01, 2025
• End date: September 30, 2026
• Field of knowledge: Biological Sciences - Biochemistry - Molecular Biology
• Agreement: GlaxoSmithKline
• Principal Investigator: Ana Marisa Chudzinski-Tavassi
• Grantee: Mariana Barbosa de Souza Rizzo
• Host Institution: Instituto Butantan. São Paulo , SP, Brazil
• Company: Secretaria da Saúde (São Paulo - Estado). Instituto Butantan
• Associated research grant: 20/13139-0 - Centre of Excellence in New Target Discovery, AP.PCPE

Abstract

INTRODUCTION: Pain is a global public health crisis, affecting approximately 20% of the adult population worldwide. Identifying and validating novel therapeutic targets is crucial for developing safer and more effective pain treatments. Venoms, rich in diverse bioactive compounds, are a promising source for this purpose. Our study investigates the antinociceptive potential of P4, a peptide derived from Lonomia obliqua caterpillar venom, using an in vitro model of human sensory-like neurons differentiated from SH-SY5Y cells. In this model, we stimulate pain-related responses using a glycated collagen matrix. We've found that P4 exerts its antinociceptive effect by decreasing the release of the pain-signaling neuropeptide substance P and increasing the release of the analgesic neuropeptide ¿-endorphin. Through chemoproteomics and bioinformatics, we've identified specific targets of P4 potentially involved in downregulating pain and inflammation. OBJECTIVES: To validate, through gene edition, one of the identified therapeutic targets potentially involved in pain and inflammation. METHODS: We used CRISPR/Cas9 to perform the target gene knockdown in SH-SY5Y cells. We will assess the functional impact of this gene editing using molecular and cellular biology approaches. EXPECTED RESULTS: The results of this research are expected to elucidate the role of our identified target in cellular nociceptive and inflammatory responses. This will not only advance our understanding of how antinociceptive peptides like P4 function but could also reveal a new, specific therapeutic target for the management of pain and inflammation, particularly in conditions like osteoarthritis.