Silvia Ghislanzoni

@istitutotumori.mi.it

PhD student, Molecular Mechanisms Department
Fondazione IRCCS Istituto Nazionale dei Tumori

Silvia Ghislanzoni


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https://researchid.co/ghislanzonisilvia
PhD student at Fondazione IRCCS Istituto Nazionale dei Tumori (INT)

EDUCATION

Master's Dedree, CTF (Medicinal Chemistry and Pharmaceutical Technology), Università degli Studi di Milano (MI, Italy)
15

Scopus Publications

Scopus Publications

  • Metabolic profiling of therapy-induced senescent cancer cells via TPEF, MALDI-MS, and RNA-sequencing
    Silvia Ghislanzoni, Federica Padelli, Matteo Niero, Alessia Bertolotti, Antonino Belfiore, Simone Torelli, Arianna Bresci, Andrea Masella, Silvia Betti, Dario Polli, Luca Agnelli, Italia Bongarzone
    Scientific Reports, 2026
    Despite advances in cancer therapies, treatment failure from resistance and recurrence remains a major clinical challenge. Therapy-induced senescence (TIS), a state of stable cell cycle arrest with sustained metabolic activity, has emerged as a driver of inflammation, tumor persistence, and relapse. However, the heterogeneity of TIS complicates its detection and targeting. Here, we applied a multi-modal strategy to characterize metabolic alterations in senescent cancer cells induced by doxorubicin or γ-irradiation across three tumor cell lines: MCF7, HeLa, and TPC-1. Mitochondrial dysfunction was assessed using MitoTracker and JC-1 staining, while two-photon excitation fluorescence (TPEF) microscopy enabled label-free visualization of intracellular NAD(P)H and FAD distribution. Lipid remodeling was evaluated by MALDI mass spectrometry imaging, and RNA sequencing was performed on control, senescent, and engulfing-senescent MCF7 cells to identify differentially expressed genes and enriched pathways. Senescent cells displayed mitochondrial dysfunction, with altered NAD(P)H/FAD distribution and decreased membrane potential. TPEF confirmed redistribution of coenzymes, reflecting redox changes. Lipidomics revealed consistent remodeling, notably involving cardiolipin precursors. Transcriptomic profiling showed engulfing-senescent MCF7 cells possess a distinct signature marked by increased lipid metabolism, endocrine signaling, and cell–cell communication. Overall, our findings reveal conserved and cell type–specific metabolic traits of TIS, highlighting metabolic vulnerabilities for senolytic intervention.
  • Estimation of biological variance in coherent Raman microscopy data of two cell lines using chemometrics
    Rajendhar Junjuri, Matteo Calvarese, MohammadSadegh Vafaeinezhad, Federico Vernuccio, Marco Ventura, Tobias Meyer-Zedler, Benedetta Gavazzoni, Dario Polli, Renzo Vanna, Italia Bongarzone, Silvia Ghislanzoni, Matteo Negro, Juergen Popp, Thomas Bocklitz
    Analyst, 2024
    Broadband Coherent Anti-Stokes Raman Scattering (BCARS) is a valuable spectroscopic imaging tool forestimating the biological variance of cells in biomedical applications.
  • Optical Diffraction Tomography and Raman Confocal Microscopy for the Investigation of Vacuoles Associated with Cancer Senescent Engulfing Cells
    Silvia Ghislanzoni, Jeon Woong Kang, Arianna Bresci, Andrea Masella, Koseki J. Kobayashi-Kirschvink, Dario Polli, Italia Bongarzone, Peter T. C. So
    Biosensors, 2023
    Wild-type p53 cancer therapy-induced senescent cells frequently engulf and degrade neighboring ones inside a massive vacuole in their cytoplasm. After clearance of the internalized cell, the vacuole persists, seemingly empty, for several hours. Despite large vacuoles being associated with cell death, this process is known to confer a survival advantage to cancer engulfing cells, leading to therapy resistance and tumor relapse. Previous attempts to resolve the vacuolar structure and visualize their content using dyes were unsatisfying for lack of known targets and ineffective dye penetration and/or retention. Here, we overcame this problem by applying optical diffraction tomography and Raman spectroscopy to MCF7 doxorubicin-induced engulfing cells. We demonstrated a real ability of cell tomography and Raman to phenotype complex microstructures, such as cell-in-cells and vacuoles, and detect chemical species in extremely low concentrations within live cells in a completely label-free fashion. We show that vacuoles had a density indistinguishable to the medium, but were not empty, instead contained diluted cell-derived macromolecules, and we could discern vacuoles from medium and cells using their Raman fingerprint. Our approach is useful for the noninvasive investigation of senescent engulfing (and other peculiar) cells in unperturbed conditions, crucial for a better understanding of complex biological processes.
  • Reduced sulfatide content in deferoxamine-induced senescent HepG2 cells
    Silvia Ghislanzoni, Gaia Martina Sarcinelli, Arianna Bresci, Francesco Manetti, Dario Polli, Antonella Tomassetti, Maria Teresa Radice, Italia Bongarzone
    International Journal of Biochemistry and Cell Biology, 2023
    Iron chelators, such as deferoxamine, exert an anticancer effect by altering the activity of biomolecules critical for regulation of the cell cycle, cell metabolism, and apoptotic processes. Thus, iron chelators are sometimes used in combination with radio- and/or chemotherapy in the treatment of cancer. The possibility that deferoxamine could induce a program of senescence similar to radio- and/or chemotherapy, fostering adaptation in the treatment of cancer cells, is not fully understood. Using established biochemical techniques, biomarkers linked to lipid composition, and coherent anti-Stokes Raman scattering microscopy, we demonstrated that hepatocellular carcinoma-derived HepG2 cells survive after deferoxamine treatment, acquiring phenotypic traits and representative hallmarks of senescent cells. The results support the view that deferoxamine acts in HepG2 cells to produce oxidative stress-induced senescence by triggering sequential mitochondrial and lysosomal dysfunction accompanied by autophagy blockade. We also focused on the lipidome of senescent cells after deferoxamine treatment. Using mass spectrometry, we found that the deferoxamine-induced senescent cells presented marked remodeling of the phosphoinositol, sulfatide, and cardiolipin profiles, which all play a central role in cell signaling cascades, intracellular membrane trafficking, and mitochondria functions. Detection of alterations in glycosphingolipid sulfate species suggested modifications in ceramide generation, and turnover is frequently described in cancer cell survival and resistance to chemotherapy. Blockade of ceramide generation may explain autophagic default, resistance to apoptosis, and the onset of senescence.
  • Full-Spectrum CARS Microscopy of Cells and Tissues with Ultrashort White-Light Continuum Pulses
    Federico Vernuccio, Renzo Vanna, Chiara Ceconello, Arianna Bresci, Francesco Manetti, Salvatore Sorrentino, Silvia Ghislanzoni, Flavia Lambertucci, Omar Motiño, Isabelle Martins, Guido Kroemer, Italia Bongarzone, Giulio Cerullo, Dario Polli
    Journal of Physical Chemistry B, 2023
    Coherent anti-Stokes Raman scattering (CARS) microscopy is an emerging nonlinear vibrational imaging technique that delivers label-free chemical maps of cells and tissues. In narrowband CARS, two spatiotemporally superimposed picosecond pulses, pump and Stokes, illuminate the sample to interrogate a single vibrational mode. Broadband CARS (BCARS) combines narrowband pump pulses with broadband Stokes pulses to record broad vibrational spectra. Despite recent technological advancements, BCARS microscopes still struggle to image biological samples over the entire Raman-active region (400–3100 cm–1). Here, we demonstrate a robust BCARS platform that answers this need. Our system is based on a femtosecond ytterbium laser at a 1035 nm wavelength and a 2 MHz repetition rate, which delivers high-energy pulses used to produce broadband Stokes pulses by white-light continuum generation in a bulk YAG crystal. Combining such pulses, pre-compressed to sub-20 fs duration, with narrowband pump pulses, we generate a CARS signal with a high (<9 cm–1) spectral resolution in the whole Raman-active window, exploiting both the two-color and three-color excitation mechanisms. Aided by an innovative post-processing pipeline, our microscope allows us to perform high-speed (≈1 ms pixel dwell time) imaging over a large field of view, identifying the main chemical compounds in cancer cells and discriminating tumorous from healthy regions in liver slices of mouse models, paving the way for applications in histopathological settings.
  • Multimodal Coherent Raman and Multiphoton Nonlinear Optical Microscopy Reveals Early Risk of Tumour Recurrence after Anticancer Therapy in Human Cells
    Arianna Bresci, Francesco Manetti, Silvia Ghislanzoni, Federico Vernuccio, Salvatore Sorrentino, Chiara Ceconello, Renzo Vanna, Italia Bongarzone, Giulio Cerullo, Dario Polli
    2023 Conference on Lasers and Electro Optics Europe and European Quantum Electronics Conference CLEO Europe Eqec 2023, 2023
    Recent studies have shown that common anticancer treatments can induce cell senescence rather than death, which is a critical phenotype governing tumour recurrence, dormancy and treatment resistance [1]. This calls for the urgent development of safe, precise, and rapid tools to unveil early evidence of such therapy-induced senescence (TIS).
  • Multimodal vibrational and multiphoton nonlinear optical microscopy as a non-invasive tool to prevent human tumor recurrence
    Arianna Bresci, Francesco Manetti, Silvia Ghislanzoni, Federico Vernuccio, Salvatore Sorrentino, Chiara Ceconello, Renzo Vanna, Italia Bongarzone, Giulio Cerullo, Dario Polli
    Proceedings of SPIE the International Society for Optical Engineering, 2023
    Recently, anticancer treatments were discovered to induce cell senescence other than death, a critical phenotype driving tumor recurrence. This calls for the development of safe, precise, and rapid tools to reveal critical therapy-induced senescence (TIS). Here, we present label-free multimodal nonlinear optical (NLO) microscopy as a powerful technique to spot early TIS. We home-built a microscope including different NLO modalities: Stimulated Raman Scattering (SRS), forward and epi-detected Coherent Anti-Stokes Raman Scattering (CARS and E-CARS), and Two-Photon Excited Fluorescence (TPEF). The infrared laser source outputs synchronized narrowband 780 nm pump pulses and 950-1050 nm tunable Stokes pulses, so to match the CH-stretching region of the Raman spectrum. Thanks to the co-registration of these NLO signals from label-free TIS cells and controls, we unveiled quantitative all-optical traits of early-stage TIS, monitored over 72 hours of treatment. TPEF from metabolic coenzymes combined with E-CARS from cardiolipin and cytochrome C indicated an shrinking of mitochondrial networks. CARS and SRS revealed lipid vesicles accumulation in cytoplasms. Nuclei enlarged irregularly, visualized via subtraction of SRS signals of proteins and lipids, and CARS from deoxyribose. We believe our results will strongly influence anticancer pre-clinical studies and translated clinical applications, constituting a quick, non-invasive, and accurate aid to expose TIS manifestation in tumors.
  • Multimodal vibrational and multiphoton nonlinear optical microscopy as a non-invasive tool to prevent human tumor recurrence
    Arianna Bresci, Francesco Manetti, Silvia Ghislanzoni, Federico Vernuccio, Salvatore Sorrentino, Chiara Ceconello, Renzo Vanna, Italia Bongarzone, Giulio Cerullo, Dario Polli
    European Conference on Biomedical Optics Ecbo 2023, 2023
    Recently, anticancer treatments were discovered to induce cell senescence other than death, a critical phenotype driving tumor recurrence. This calls for the development of safe, precise, and rapid tools to reveal critical therapy-induced senescence (TIS). Here, we present label-free multimodal nonlinear optical (NLO) microscopy as a powerful technique to spot early TIS. We home-built a microscope including different NLO modalities: Stimulated Raman Scattering (SRS), forward and epi-detected Coherent Anti-Stokes Raman Scattering (CARS and E-CARS), and Two-Photon Excited Fluorescence (TPEF). The infrared laser source outputs synchronized narrowband 780 nm pump pulses and 950-1050 nm tunable Stokes pulses, so to match the CH-stretching region of the Raman spectrum. Thanks to the co-registration of these NLO signals from label-free TIS cells and controls, we unveiled quantitative all-optical traits of early-stage TIS, monitored over 72 hours of treatment. TPEF from metabolic coenzymes combined with E-CARS from cardiolipin and cytochrome C indicated an shrinking of mitochondrial networks. CARS and SRS revealed lipid vesicles accumulation in cytoplasms. Nuclei enlarged irregularly, visualized via subtraction of SRS signals of proteins and lipids, and CARS from deoxyribose. We believe our results will strongly influence anticancer pre-clinical studies and translated clinical applications, constituting a quick, non-invasive, and accurate aid to expose TIS manifestation in tumors.
  • Deep ensemble learning and transfer learning methods for classification of senescent cells from nonlinear optical microscopy images
    Salvatore Sorrentino, Francesco Manetti, Arianna Bresci, Federico Vernuccio, Chiara Ceconello, Silvia Ghislanzoni, Italia Bongarzone, Renzo Vanna, Giulio Cerullo, Dario Polli
    Frontiers in Chemistry, 2023
    The success of chemotherapy and radiotherapy anti-cancer treatments can result in tumor suppression or senescence induction. Senescence was previously considered a favorable therapeutic outcome, until recent advancements in oncology research evidenced senescence as one of the culprits of cancer recurrence. Its detection requires multiple assays, and nonlinear optical (NLO) microscopy provides a solution for fast, non-invasive, and label-free detection of therapy-induced senescent cells. Here, we develop several deep learning architectures to perform binary classification between senescent and proliferating human cancer cells using NLO microscopy images and we compare their performances. As a result of our work, we demonstrate that the most performing approach is the one based on an ensemble classifier, that uses seven different pre-trained classification networks, taken from literature, with the addition of fully connected layers on top of their architectures. This approach achieves a classification accuracy of over 90%, showing the possibility of building an automatic, unbiased senescent cells image classifier starting from multimodal NLO microscopy data. Our results open the way to a deeper investigation of senescence classification via deep learning techniques with a potential application in clinical diagnosis.
  • Multimodal multiphoton and vibrational microscopy reveals early therapy-induced senescence in human tumors: a non-invasive tool to prevent the risk of cancer relapse
    Arianna Bresci, Francesco Manetti, Silvia Ghislanzoni, Federico Vernuccio, Salvatore Sorrentino, Chiara Ceconello, Renzo Vanna, Italia Bongarzone, Giulio Cerullo, Dario Polli
    Progress in Biomedical Optics and Imaging Proceedings of SPIE, 2023
    Recent studies have shown that common anticancer treatments can induce cell senescence rather than death, a critical phenotype governing tumor recurrence. This calls for the urgent development of safe, precise, and quick tools to unveil critical Therapy-Induced Senescence (TIS). Merging different coherent Raman and multiphoton techniques, we present label-free multimodal nonlinear optical (NLO) microscopy as a powerful tool to spot early TIS. We home-built a microscope including different NLO modalities: Stimulated Raman Scattering (SRS), forward and epi-detected Coherent Anti-Stokes Raman Scattering (CARS and E-CARS), and Two-Photon Excited Fluorescence (TPEF). The infrared laser source outputs synchronized narrowband 780 nm pump pulses and 950-1050 nm tunable Stokes pulses, so to match the CH-stretching region of the Raman spectrum. Thanks to the co-registration of these diverse techniques applied on label-free TIS cells and controls, we exposed quantitative hallmarks of early TIS, confirmed by comparing different optical signals monitored over 72 hours of treatment. TPEF from metabolic coenzymes combined with E-CARS from cardiolipin and cytochrome C indicated an early shrinking of mitochondria. CARS and SRS revealed lipid vesicles overproduction and accumulation. Nuclei enlarged irregularly, visualized via subtraction of SRS signals of proteins and lipids, and CARS from deoxyribose. We consider our results will strongly influence anticancer pre-clinical studies and translated clinical applications, helping to identify quickly, non-invasively, and quantitatively TIS in human tumors.
  • Deep Ensemble Learning and Transfer Learning Methods for Classification of Senescent Cells in Nonlinear Optical Microscopy Images
    Francesco Manetti, Salvatore Sorrentino, Arianna Bresci, Federico Vernuccio, Chiara Ceconello, Silvia Ghislanzoni, Italia Bongarzone, Renzo Vanna, Giulio Cerullo, Dario Polli
    2023 Conference on Lasers and Electro Optics Europe and European Quantum Electronics Conference CLEO Europe Eqec 2023, 2023
  • Noninvasive morpho-molecular imaging reveals early therapy-induced senescence in human cancer cells
    Arianna Bresci, Jeong Hee Kim, Silvia Ghislanzoni, Francesco Manetti, Lintong Wu, Federico Vernuccio, Chiara Ceconello, Salvatore Sorrentino, Ishan Barman, Italia Bongarzone, Giulio Cerullo, Renzo Vanna, Dario Polli
    Science Advances, 2023
  • Plug-and-play stimulated Raman microscopy system for broadband coherent vibrational imaging
    Francesco Crisafi, Benedetta Talone, Andrea Ragni, Gabriele Di Noia, Mujeeb Rahman, Jing He, Jeremiah Marcellino, Goutam Kar, Yarjan Samad, Boyang Mao, Renzo Vanna, Franziska Hoffmann, Orlando Guntinas-Lichius, Silvia Ghislanzoni, Italia Bongarzone, Sze Yun Set, Andrea C. Ferrari, Giulio Cerullo, Matteo Negro
    2023 Conference on Lasers and Electro Optics Europe and European Quantum Electronics Conference CLEO Europe Eqec 2023, 2023
  • Multimodal vibrational and multi-photon nonlinear optical microscopy to assess chemotherapy-induced senescence in human cancer cells
    Optics Infobase Conference Papers, 2022
  • Verbascoside protects pancreatic β-cells against er-stress
    Alessandra Galli, Paola Marciani, Algerta Marku, Silvia Ghislanzoni, Federico Bertuzzi, Raffaella Rossi, Alessia Di Giancamillo, Michela Castagna, Carla Perego
    Biomedicines, 2020