PhD Student

PhD program:: XXXVII
phone: 0649920963
building: Dipartimento di Scienze Biochimiche "A. Rossi Fanelli"
room: S32

supervisor: Alessio Paone
advisor: Francesca Cutruzzolà
co-supervisor: Francesca Cutruzzolà

Research: Influence of the tumor microenvironment on the formation of brain metastases by breast cancer cells

Title of the research project: Influence of the tumor microenvironment on the formation of brain metastases by breast cancer cells

Thematic area: Molecular Biology and Biochmestry


Brief resume:
Master’s degree in Medical and Pharmaceutical Biotecnology (LM-9), at University of Pavia , 28/04/2021, final mark: 110 cum laude /110.
Thesis title: “Characterization of C. elegans transngenic strain overexpressing b2-microglobulin's gene ".
- Bachelor’s degree in Biological Sciences (L-13), at University of Tuscia, 28/02/2019, final mark: 108/110.
Thesis title: “ Panoramic of prion protein Prpc and its role in prion desease"
- Classic High School, July 2015, liceo classico “Vitruvio Pollione" Formia (LT), final mark: 80/100

Brief description of your project:
Background: Breast cancer, a devastating disease affecting millions of women worldwide, often leads to unfortunate outcomes due to the formation of brain metastases, posing a significant challenge. Although the association between cytokines produced by cancer cells and metastases formation has been proposed long time ago, no specific cytokine has been identified as responsible for allowing tumor cells to enter the brain parenchyma. In addition, the metabolic remodeling induced by cytokines can result in brain accumulation of glutamate, which may act as a propellant to help tumor cells infiltrate the organ.

Aim: we plan to understand the role of neurotransmitter amino acids, with focus on glutamate, in the mechanism of extravasation of breast cancer cells and we wish to uncover any possible connection and synergistic effects between the mechanisms that regulate cytokine-induced effects and neurotransmitter amino acid function within the unique microenvironment of the brain's blood capillaries to further clarify their role in the mechanism of extravasation.

Methodologies: I acquired different techniques in cell biology and analysis of cancer metabolism, using several approaches such as DNA Cell transfection, viability test through Trypan blue assays, qPCR for quantify gene expression, Migration test using Boyden chamber, Permeability test and Seahorse analysis. Moreover, I am also able to perform molecular biology experiments as PCR, DNA/RNA extraction, Western Blot, Electrophoresis.

Results: Our research utilizes two triple-negative breast cancer cell types: MDA MB 231 and MDA MB 231 BR. The latter one has a metastatic profile compared to 231 cells. The results demonstrated that in 231 BR cells there is an increase in NF-KB activity compared to the parental cell lines. Therefore, we investigated how the most common proteins of NF-KB pathway, as for example IKKa, IKKβ, IKBa, are modulated in our cellular model. We demonstrated that in 231 BR cells the higher IKKβ protein expression leads to an IKBa degradation causing an increase in NF-KB activity, otherwise in 231 BR cells the lower IKKa protein expression leads to an increase in NF-KB activity compared to 231 cells in which we observed an opposite trend. Moreover, we investigated the presence of cytokines in the conditioned medium of 231 and 231 Br and we observed significantly different pattern of cytokines in the conditioned medium of 231 Br cells compared to parental cells. To study the effect of these cytokines, we focus on four cytokines that are upregulated in 231 BR conditioned medium that have the same receptor CXCR1 and CXCR2. We performed a permeability test: at the top of the porous filter we seeded the blood brain barrier cells (BBB cells), upon the BBB cells we added the conditioned medium of 231 and 231 Br with and without the cytokines receptor inhibitor. We have observed that the conditioned medium from MDA-MB-231Br cells is more efficient in increasing the permeability compared to the conditioned medium from parental MDA- MB-231 cells. Instead, with the cytokine receptor inhibitor there is a decrease in the permeability in the presence of both conditioned medium of 231 and 231 Br, suggesting that these cytokines have a necessary role in facilitating the opening of the blood-brain barrier. Moreover, we measured the metabolism of 231 and 231 Br by performing Seahorse experiments, treating both cells, 231 and 231 Br, with the cytokine receptor inhibitor (CXCR1 and CXCR2) and only 231 cells with the upregulated cytokines found in the 231 Br conditioned medium: we observed that there is a decrease in the oxygen consumption rate (OCR) and in the extracellular acidification rate (ECAR) in cells treated with the cytokine receptor inhibitor and a small increase of both OCR and ECAR value in cells treated with cytokines. This suggests that the cytokines could sustain the metabolism of 231 and 231 Br cells. In this context, the role of glutamate is fundamental because is a significant metabolite on the brain side of the blood brain barrier and we have observed that metastatic breast cancer cells detect and import glutamate via EAAT1 transporter. Our results suggest that glutamate enhances the migratory ability of metastatic breast cancer cells when they attempt to cross the BBB and we have preliminary data demonstrated that probably the presence of cytokines released by breast cancer cells that metastasize in the brain, reduced the glutamate uptake by astrocytes, determining an accumulation of glutamate in the brain that can then pass from brain to blood where it could be used by cancer cells.

Formal participation to funded research projects:
Title: Dissecting Serine Hydroxymethyltransferase functions to target lung cancer metabolic reprogramming
Years: 2020-2024
Funding agency: Progetto Associazione Italiana Ricerca sul Cancro (AIRC) IG 23125
PI: Francesca Cutruzzolà

Title: RNA as a new player in the allosteric control of cellular metabolism: role of riboregulation in cancer
Years: 2023-2025
Funding agency: Progetto PRIN (MUR) prot. 20228395KW
PI: Francesca Cutruzzolà

Title: A two-pronged approach to target the Aurora-A/N-Myc complex in MYCN-amplified Neuroblastoma
Years: 2022-2024
Funding agency: Progetto Ateneo 2022 RM1221815D52AB32
PI: Alessandro Paiardini

-From Ramon y cajal to cryoem course for adavanced microscopy (Speaker: Silvia D’Angelantonio, Egle De Stefano, Giorgio Giardina)
-Biotecnologie e Sicurezza in Laboratorio (Speaker: Elena Sturchio, Miriam Zanellato)

-Engineering Proteins To Boost LTP And Memory (Speaker: Cristian Ripoli )
-Application of deep learning to the protein structure prediction: the tale of a “gigantic leap”
(Speaker: Anna Marabotti)
-Deciphering self-renewal traits in epidermal stem cells (Speaker: Elena Enzo)
-Insulin Signaling in Alzheimer’s Disease Brain and Models Thereof (Speaker: Eugenio Barone)
-Educating tutor-associate macrophages under metabolic stress (Speaker: Ping-Chih Ho)
-Synthetic Biology: what, why and how (Speaker: Velia Siciliano)
-Protecting our genome: mechanisms to maintain DNA repeats stability in human cells (Speaker: Simona Giunta)
-Regulation of enzymatic activity mediated by liquid-liquid phase separation (Speaker: Mirco Dindo)
-The chaperonin GroEL nano-machine: allostery and function (Speaker: Amnon Horowitz)
The axonal transport machinery and its dysfunctions in neurodegenerative diseases (Speaker: Giampietro Schiavo)
-Riboregulation from bacteria to eukaryotes: mechanisms and challenges (Speaker: Matthias Hentze, Ina Huppertz, Ben Luisi, Gian Gaetano Tartaglia, Francesca Cutruzzolà)

-The 47th FEBS Congress- Flask Talk and Poster Presentation "Influence of the tumor microenvironment on the formation of brain metastases by breast cancer cells"
-3rd Workshop SIB "Understanding cancer metabolism: exploring tumor heterogeneity to advance cancer therapy"- Oral Presentation " Role of amino acids metabolism in the metastatic potential of breast and lung cancer cells"

Research products

11573/1702420 - 2024 - Beyond the barrier: the immune-inspired pathways of tumor extravasation
Di Russo, S.; Liberati, F. R.; Riva, A.; Di Fonzo, F.; Macone, A.; Giardina, G.; Arese, M.; Rinaldo, S.; Cutruzzola, F.; Paone, A. - 01g Articolo di rassegna (Review)
paper: CELL COMMUNICATION AND SIGNALING (BioMed Central, London) pp. 104- - issn: 1478-811X - wos: WOS:001160608800001 (0) - scopus: 2-s2.0-85184788152 (0)

11573/1707844 - 2024 - Combined Delivery of miR-15/16 through Humanized Ferritin Nanocages for the Treatment of Chronic Lymphocytic Leukemia
Liberati, Francesca Romana; Di Russo, Sara; Barolo, Lorenzo; Peruzzi, Giovanna; Farina, Maria Vittoria; Spizzichino, Sharon; Di Fonzo, Federica; Quaglio, Deborah; Pisano, Luca; Botta, Bruno; Giorgi, Alessandra; Boffi, Alberto; Cutruzzola, Francesca; Paone, Alessio; Baiocco, Paola - 01a Articolo in rivista
paper: PHARMACEUTICS (Basel: MDPI, 2009-) pp. - - issn: 1999-4923 - wos: (0) - scopus: (0)

© Università degli Studi di Roma "La Sapienza" - Piazzale Aldo Moro 5, 00185 Roma