New compounds with anti-cancer activity from microorganisms from MicroBioBank: cell panel screnn, mechanism of action and biosynthetic production

• Grant number: 24/22909-5
• Support Opportunities: Scholarships in Brazil - Doctorate (Direct)
• Start date: March 01, 2025
• End date: July 31, 2028
• Field of knowledge: Interdisciplinary Subjects
• Principal Investigator: Daniela Barretto Barbosa Trivella
• Grantee: Paola Ferrari
• Host Institution: Centro Nacional de Pesquisa em Energia e Materiais (CNPEM). Ministério da Ciência, Tecnologia e Inovação (Brasil). Campinas , SP, Brazil
• Associated research grant: 21/12590-3 - Microbial biobanks: promoting innovations for scientific research and technological development (MicroBioBank), AP.TEM

Abstract

Cancer is a group of diseases that constitute unmet medical needs, or whose treatments are suboptimal. The World Health Organization (WHO) reports that approximately 20 million people were diagnosed with cancer in 2020 and 10 million died in 2021. Natural products represent more than 65% of the source of molecules already developed with anticancer action, acting on the most diverse types of cancer. In this context, MicroBioBank researchers intend to explore the wealth of molecules in the biobanks associated with the Thematic project in order to advance the discovery of new bioactive natural products with anticancer action. The Direct Doctorate project presented here aims to screen the MicroBioBank library using a phenotypic assay in order to find and deepen knowledge about new natural products with selective cytotoxic action for cancer cells. The proposed activities also aim to support the bioguided isolation activities to be carried out in the context of the Thematic, evaluate the mechanism of action of the bioactive natural product in cells and its production by biotechnological route. Initially, HaCat cells (non-cancerous, used as a reference for normal cells) and HCT-116 cells (colon cancer, used as a reference for cancer cells) will be used to screen the MicroBioBank library. Cytotoxicity values (quantified through the IC50 parameter) and the selectivity index will be used to compare the cytotoxicity of the natural samples and their action on HCT-116 cells (target) versus HaCat cells (control). The selected hits will be evaluated in an expanded cancer cell panel, including breast cancer and leukemia. The hits will be dereplicated by the Thematic team (see mass spectrometry analysis and identification of bioactive natural products - a PhD scholarship in chemistry also requested in the project), whose activity will result in the selection of up to 8 prioritized fungi for bioguided isolation of the natural product with anticancer action. The biological evaluation of these purified samples is also the scope of this work plan. The natural products isolated by the Thematic team will be evaluated for their mechanisms of action in multiparametric cellular assays, which assess toxicity, mitochondrial activity, oxidative stress, membrane damage, apoptosis and reticulum stress, the latter two mechanisms being the most desired for selective anticancer action. In parallel with the bioguided isolation activities and investigation of the mechanisms of action, the genomes of the prioritized fungi will be sequenced and mined by the doctoral student, where it is expected to indicate biosynthetic gene clusters for the production of new bioactive natural products with anticancer action. Genome mining will also be able to indicate the mechanism of action of the bioactive natural products (e.g.: presence of resistance genes in the selected biosynthetic cluster), further supporting the evaluation of the mechanism of action of these new natural products with anticancer action. A biosynthetic gene cluster will be cloned in a heterologous system for the production of the bioactive natural product, confirmation of its chemical structure and evaluation of the viability of production by biosynthetic route. (AU)