Project description
Single-cell technology helps to define sources of nucleotide de novo synthesis
Nucleotide de novo synthesis (DNS) is essential for cell proliferation, and its disruption is detrimental to rapidly multiplying cancer cells. Anti-nucleotide therapy was one of the first approaches to treat cancer, but DNS can be bypassed by the uptake of extracellular nucleotides or through the recycling pathways, limiting therapy efficacy. The cellular sources of nucleotides in normal tissue and tumours in vivo have not yet been adequately characterised; this is the main goal of the EU-funded MetaCross project, in addition to understanding the adaptations to DNS blockade in cancer. The research will involve an integrative in vivo approach using single-cell technology combined with single-cell omics analysis, advanced bioinformatics and state-of-the-art mouse models.
Objective
DNA synthesis is essential for cell proliferation. Nucleotides, the basic building blocks of nucleic acids, are made by nucleotide de novo synthesis (DNS), and DNS disruption is detrimental to rapidly proliferating cancer cells. Established >70 years ago, anti-nucleotide therapy was one of the first approaches to treat cancer, but it suffers high rate of resistance and relapse. DNS can be bypassed by an uptake of extracellular nucleotides or by recycling in salvage pathways, possibly a reason for limited efficacy of anti-nucleotide therapy. To date, the cellular sources of nucleotides in normal tissue and in tumors in vivo remain poorly characterized. The central goal of this project is to define these nucleotide sources, understand the intercellular metabolic crosstalk of nucleotides in tumors, and characterize the adaptations to DNS blockade in cancer and stromal cells. To reach these goals, I will use the totally new perspective brought by the single cell technology and combine my expertise in single cell omics and metabolism with the state-of-the-art mouse models and advanced bioinformatics available at the host institute. I propose an integrative in vivo approach using single cell RNA-sequencing, which in combination with genetic interventions will allow me to resolve dynamic expression profiles of individual cell types. I will use inducible mouse models to selectively disable DNS in the stroma (lungs of a host animal) and in cancer cells (orthotopic tumors from syngeneic DNS-deficient lung cancer cells) to generate tumors fully relying on nucleotides from internal or external sources, respectively. In parallel, I will perform in vivo CRISPR screen to identify genes whose lack represents a targetable metabolic dependency of DNS-disabled cancer cells. This innovative approach will shed novel insights into organization of tumor metabolic homeostasis and identify new targets with the potential to make major breakthrough in anti-nucleotide intervention in cancer.
Fields of science
- natural sciencesbiological sciencesbiochemistrybiomoleculesnucleic acids
- medical and health sciencesclinical medicineoncologylung cancer
- natural sciencesbiological sciencesgeneticsnucleotides
- medical and health sciencesmedical biotechnologycells technologies
- medical and health sciencesbasic medicinephysiologyhomeostasis
Programme(s)
Funding Scheme
MSCA-IF - Marie Skłodowska-Curie Individual Fellowships (IF)Coordinator
252 50 Vestec
Czechia