Omega-3 fatty acid DHA induces ferroptosis in colorectal cancer patient-derived organoids and drug-tolerant cells Laura di Blasio, Marianela Vara-Messler, Barbara Peracino, Elena Masti, Vanesa Cepas-López, et al. Cell Death and Disease, 2026 Several epidemiological and preclinical studies suggest that omega-3 (n-3) polyunsaturated fatty acids (PUFAs) exert anticancer activity at multiple stages of colorectal cancer (CRC) progression. However, inconsistent clinical evidence and the lack of a clearly defined molecular mechanism underlying the antitumor effects of n-3 PUFAs have raised doubts about their efficacy as anticancer therapies. To address these issues, we investigated the effects of the n-3 PUFA docosahexaenoic acid (DHA) in a collection of CRC patient-derived tumor organoids (PDTOs), a powerful platform for functional analysis of patient-specific tumors. DHA treatment markedly reduced CRC cell viability in a time- and concentration-dependent manner without inducing apoptosis. CRC-derived PDTOs exhibited pronounced sensitivity to DHA, irrespective of KRAS or TP53 mutational status, whereas organoids from normal colon tissue were less affected. Mechanistically, DHA induced ferroptosis in both CRC cells and PDTOs, as evidenced by lipid peroxide accumulation and partial rescue by ferroptosis inhibitors. Fluorescently labeled DHA localized predominantly to the endoplasmic reticulum and mitochondria, where it promoted oxidative stress. Moreover, DHA impaired the regrowth of oxaliplatin-tolerant persister cells and enhanced oxaliplatin efficacy in sequential treatment models. Together, these findings indicate that exploiting the intrinsic oxidative vulnerability of cancer cells with DHA may represent a promising, low-toxicity strategy to enhance chemotherapy efficacy and target drug-tolerant persister cells in colorectal cancer.
Two-fluid dynamics and micron-thin boundary layers shape cytoplasmic flows in early Drosophila embryos Claudio Hernández-López, Alberto Puliafito, Yitong Xu, Ziqi Lu, Stefano Di Talia, et al. Proceedings of the National Academy of Sciences of the United States of America, 2023 Cytoplasmic flows are widely emerging as key functional players in development. In early Drosophila embryos, flows drive the spreading of nuclei across the embryo. Here, we combine hydrodynamic modeling with quantitative imaging to develop a two-fluid model that features an active actomyosin gel and a passive viscous cytosol. Gel contractility is controlled by the cell cycle oscillator, the two fluids being coupled by friction. In addition to recapitulating experimental flow patterns, our model explains observations that remained elusive and makes a series of predictions. First, the model captures the vorticity of cytosolic flows, which highlights deviations from Stokes’ flow that were observed experimentally but remained unexplained. Second, the model reveals strong differences in the gel and cytosol motion. In particular, a micron-sized boundary layer is predicted close to the cortex, where the gel slides tangentially while the cytosolic flow cannot slip. Third, the model unveils a mechanism that stabilizes the spreading of nuclei with respect to perturbations of their initial positions. This self-correcting mechanism is argued to be functionally important for proper nuclear spreading. Fourth, we use our model to analyze the effects of flows on the transport of the morphogen Bicoid and the establishment of its gradients. Finally, the model predicts that the flow strength should be reduced if the shape of the domain is more round, which is experimentally confirmed in Drosophila mutants. Thus, our two-fluid model explains flows and nuclear positioning in early Drosophila , while making predictions that suggest novel future experiments.
A modified fluctuation-test framework characterizes the population dynamics and mutation rate of colorectal cancer persister cells Mariangela Russo, Simone Pompei, Alberto Sogari, Mattia Corigliano, Giovanni Crisafulli, et al. Nature Genetics, 2022 Compelling evidence shows that cancer persister cells represent a major limit to the long-term efficacy of targeted therapies. However, the phenotype and population dynamics of cancer persister cells remain unclear. We developed a quantitative framework to study persisters by combining experimental characterization and mathematical modeling. We found that, in colorectal cancer, a fraction of persisters slowly replicates. Clinically approved targeted therapies induce a switch to drug-tolerant persisters and a temporary 7- to 50-fold increase of their mutation rate, thus increasing the number of persister-derived resistant cells. These findings reveal that treatment may influence persistence and mutability in cancer cells and pinpoint inhibition of error-prone DNA polymerases as a strategy to restrict tumor recurrence.
Recruitment, infiltration and cytotoxicity of hla-independent killer lymphocytes in three-dimensional melanoma models Ilenia Iaia, Loretta Gammaitoni, Giulia Cattaneo, Lidia Giraudo, Chiara Donini, et al. Cancers, 2021 Cancer adoptive cell therapy (ACT) with HLA-independent tumor killer lymphocytes is a promising approach, with intrinsic features potentially addressing crucial tumor-escape mechanisms of checkpoint inhibitors. Cytokine-induced Killer (CIK) and Natural Killer (NK) lymphocytes share similar tumor-killing mechanisms, with preclinical evidence of intense activity against multiple solid tumors and currently testing in clinical studies. To improve the effective clinical translation of such ACT approaches, several fundamental questions still need to be addressed within appropriate preclinical contexts, capable of overcoming limitations imposed by most traditional two-dimensional assays. Here, we developed a novel experimental approach to explore, dissect, and visualize the interactions of CIK and NK lymphocytes with melanoma tumors in vitro in 3D. Primary melanoma cells were assembled into small tumors that were dispersed in a 3D matrix and challenged with patient-derived CIK or the NK-92 cell line. By means of imaging-based methods, we reported, visualized, and quantitatively measured the recruitment of CIK and NK on the 3D targets, their infiltration, and cytotoxic activity. Our results support the effective tumor recruitment and tumor infiltration by CIK and NK. Such features appeared dependent on the specific geometric aspects of the environment but can be explained in terms of directional migration toward the tumor, without invoking major feedback components. Overall, our 3D platform allows us to monitor the processes of tumor recruitment, infiltration, and killing by means of live measurements, revealing important kinetic aspects of ACT with CIK and NK against melanoma.
Activation of RSK by phosphomimetic substitution in the activation loop is prevented by structural constraints Desiana Somale, Giovanna Di Nardo, Laura di Blasio, Alberto Puliafito, Marianela Vara-Messler, et al. Scientific Reports, 2020 The activation of the majority of AGC kinases is regulated by two phosphorylation events on two conserved serine/threonine residues located on the activation loop and on the hydrophobic motif, respectively. In AGC kinase family, phosphomimetic substitutions with aspartate or glutamate, leading to constitutive activation, have frequently occurred at the hydrophobic motif site. On the contrary, phosphomimetic substitutions in the activation loop are absent across the evolution of AGC kinases. This observation is explained by the failure of aspartate and glutamate to mimic phosphorylatable serine/threonine in this regulatory site. By detailed 3D structural simulations of RSK2 and further biochemical evaluation in cells, we show that the phosphomimetic residue on the activation loop fails to form a critical salt bridge with R114, necessary to reorient the αC-helix and to activate the protein. By a phylogenetic analysis, we point at a possible coevolution of a phosphorylatable activation loop and the presence of a conserved positively charged amino acid on the αC-helix. In sum, our analysis leads to the unfeasibility of phosphomimetic substitution in the activation loop of RSK and, at the same time, highlights the peculiar structural role of activation loop phosphorylation.
Omega-3 fatty acid DHA induces ferroptosis in colorectal cancer patient-derived organoids and drug-tolerant cells L Di Blasio, M Vara-Messler, B Peracino, E Masti, V Cepas-López, ... Cell Death & Disease , 2026 2026
A cancer cell intrinsic program for matrix remodeling and CAF recruitment in slow growing, poor prognosis colorectal cancers. C Isella, A Cassisa, C Petti, AA Ulla, J Zhou, C Leonardi, R Porporato, ... Cancer Research 86 (7_Supplement), 776-776 , 2026 2026
Omega-3 Fatty Acid DHA Induces Ferroptosis in Colorectal Cancer Patient-Derived Organoids and Drug-Tolerant Cells L Primo, L di Blasio, M Vara-Messler, B Peracino, E Masti, VC López, ... 2026
Exploring redox dynamics in distinct cell populations through a facs-based approach using cancer patient-derived organoids and compartment-specific genetically-encoded redox … V Cepas-López, F Galvagno, M Miglio, L di Blasio, LP Roma, A Puliafito, ... Free Radical Biology and Medicine 233, S65 , 2025 2025
Single-cell analysis and cytokines modulation of intra-tumor subpopulations in colorectal cancer tumoroids SJ Fletcher, I Catalano, E Grassi, S Borgato, B Peracino, L Primo, ... Cancer Research 85 (8_Supplement_1), 2665-2665 , 2025 2025
Inference of lineage hierarchies, growth and drug response mechanisms in cancer cell populations without tracking A Piras, F Galvagno, L Pizzini, E Grassi, A Bertotti, L Primo, A Celani, ... bioRxiv, 2025.08. 14.669184 , 2025 2025
Three-dimensional dynamics of mesothelin-targeted CAR. CIK lymphocytes against ovarian cancer peritoneal carcinomatosis F Galvagno, V Leuci, A Massa, C Donini, R Rotolo, S Capellero, ... Cancer Immunology, Immunotherapy 74 (1), 6 , 2024 2024 Citations: 4
XENTURION is a population-level multidimensional resource of xenografts and tumoroids from metastatic colorectal cancer patients SM Leto, E Grassi, M Avolio, V Vurchio, F Cottino, M Ferri, ER Zanella, ... Nature Communications 15 (1), 7495 , 2024 2024 Citations: 17
Studying redox biology of cancer by using patient-derived organoids (PDOs) and compartment-specific genetically-encoded redox biosensors V Cepas-Lopez, M Miglio, F Galvagno, L Di Blasio, A Puliafito, LP Roma, ... FEBS OPEN BIO 14, 208-209 , 2024 2024
Defining single cell redox dynamics by using breast cancer patient-derived organoids (PDOs) and compartment-specific genetically-encoded redox biosensors V Cepas-López, M Miglio, L di Blasio, LP Roma, A Puliafito, L Primo Free Radical Biology and Medicine 218, 7 , 2024 2024
Molecular and therapeutic characterization of a large-scale collection of metastatic colorectal cancer patient-derived xenografts and matched organoids for translational oncology SM Leto, E Grassi, M Avolio, V Vurchio, F Cottino, M Ferri, ER Zanella, ... Cancer Research 84 (6_Supplement), 219-219 , 2024 2024
Unravelling intra-tumor phenotypic heterogeneity in colorectal cancer organoids SJ Fletcher, I Catalano, E Grassi, S Borgato, B Peracino, E Piretto, ... Cancer Research 84 (6_Supplement), 6946-6946 , 2024 2024
Two-fluid dynamics and micron-thin boundary layers shape cytoplasmic flows in early Drosophila embryos C Hernández-López, A Puliafito, Y Xu, Z Lu, S Di Talia, M Vergassola Proceedings of the National Academy of Sciences 120 (44), e2302879120 , 2023 2023 Citations: 17
XENTURION, a multidimensional resource of xenografts and tumoroids from metastatic colorectal cancer patients for population-level translational oncology SM Leto, E Grassi, M Avolio, V Vurchio, F Cottino, M Ferri, ER Zanella, ... bioRxiv, 2023.07. 10.548375 , 2023 2023 Citations: 3
Patient-derived organoids (PDOs) and compartment-specific genetically-encoded redox sensors as a novel tool to study the redox biology of human diseases V Cepas-López, LP Roma, A Puliafito, L Primo Free Radical Biology and Medicine 201, 42 , 2023 2023
Anti-mesothelin CAR. CIK lymphocytes are effective against ovarian cancer in 3D models of peritoneal carcinomatosis F Galvagno, V Leuci, C Donini, A Massa, R Rotolo, S Capellero, ... Cancer Research 83 (7_Supplement), 3194-3194 , 2023 2023
Link to this full text: http://hdl. handle. net/2318/1559469 B Lupo, J Vialard, F Sassi, P Angibaud, A Puliafito, E Pupo, L Lanzetti, ... 2023
Collective directional migration drives the formation of heteroclonal cancer cell clusters M Palmiero, I Cantarosso, L di Blasio, V Monica, B Peracino, L Primo, ... Molecular Oncology , 2023 2023 Citations: 9
A modified fluctuation-test framework characterizes the population dynamics and mutation rate of colorectal cancer persister cells M Russo, S Pompei, A Sogari, M Corigliano, G Crisafulli, A Puliafito, ... Nature genetics 54 (7), 976-984 , 2022 2022 Citations: 69
Cullin-5 mutants reveal collective sensing of the nucleocytoplasmic ratio in Drosophila embryogenesis L Hayden, A Chao, VE Deneke, M Vergassola, A Puliafito, S Di Talia Current Biology 32 (9), 2084-2092. e4 , 2022 2022 Citations: 19
MOST CITED SCHOLAR PUBLICATIONS
Collective and single cell behavior in epithelial contact inhibition A Puliafito, L Hufnagel, P Neveu, S Streichan, A Sigal, DK Fygenson, ... Proceedings of the National Academy of Sciences 109 (3), 739-744 , 2012 2012 Citations: 552
PDK1: At the crossroad of cancer signaling pathways PA Gagliardi, A Puliafito, L Primo Seminars in cancer biology 48, 27-35 , 2018 2018 Citations: 214
Self-Organized Nuclear Positioning Synchronizes the Cell Cycle in Drosophila Embryos VE Deneke, A Puliafito, D Krueger, AV Narla, A De Simone, L Primo, ... Cell 177 (4), 925-941. e17 , 2019 2019 Citations: 160
Endothelial podosome rosettes regulate vascular branching in tumour angiogenesis G Seano, G Chiaverina, PA Gagliardi, L di Blasio, A Puliafito, C Bouvard, ... Nature cell biology 16 (10), 931-941 , 2014 2014 Citations: 141
Heterogeneities in Nanog expression drive stable commitment to pluripotency in the mouse blastocyst P Xenopoulos, M Kang, A Puliafito, S Di Talia, AK Hadjantonakis Cell reports 10 (9), 1508-1520 , 2015 2015 Citations: 140
TFEB controls vascular development by regulating the proliferation of endothelial cells G Doronzo, E Astanina, D Corà, G Chiabotto, V Comunanza, A Noghero, ... The EMBO journal 38 (3), e98250 , 2019 2019 Citations: 104
PI3K/mTOR inhibition promotes the regression of experimental vascular malformations driven by PIK3CA -activating mutations L Di Blasio, A Puliafito, PA Gagliardi, V Comunanza, D Somale, ... Cell death & disease 9 (2), 45 , 2018 2018 Citations: 103
Multicolor Cell Barcoding Technology for Long-Term Surveillance of Epithelial Regeneration in Zebrafish CH Chen, A Puliafito, BD Cox, L Primo, Y Fang, S Di Talia, KD Poss Developmental Cell 36 (6), 668-680 , 2016 2016 Citations: 97
Polymers in linear shear flow: a numerical study A Celani, A Puliafito, K Turitsyn EPL (Europhysics Letters) 70 (4), 464-470 , 2005 2005 Citations: 83
Dynamic interplay between pericytes and endothelial cells during sprouting angiogenesis G Chiaverina, L di Blasio, V Monica, M Accardo, M Palmiero, B Peracino, ... Cells 8 (9), 1109 , 2019 2019 Citations: 81
Dynamic Interplay between Pericytes and Endothelial Cells during Sprouting Angiogenesis G Chiaverina, L di Blasio, V Monica, M Accardo, M Palmiero, B Peracino, ... Cells 8 (9), 1109 , 2019 2019 Citations: 81
Serine/Threonine Kinase 3-Phosphoinositide-Dependent Protein Kinase-1 (PDK1) as a Key Regulator of Cell Migration and Cancer Dissemination L Di Blasio, PA Gagliardi, A Puliafito, L Primo Cancers 9 (3), 25 , 2017 2017 Citations: 79
A modified fluctuation-test framework characterizes the population dynamics and mutation rate of colorectal cancer persister cells M Russo, S Pompei, A Sogari, M Corigliano, G Crisafulli, A Puliafito, ... Nature genetics 54 (7), 976-984 , 2022 2022 Citations: 69
Colorectal cancer residual disease at maximal response to EGFR blockade displays a druggable Paneth cell–like phenotype B Lupo, F Sassi, M Pinnelli, F Galimi, ER Zanella, V Vurchio, G Migliardi, ... Science translational medicine 12 (555), eaax8313 , 2020 2020 Citations: 69
Polarity, cell division, and out-of-equilibrium dynamics control the growth of epithelial structures B Cerruti, A Puliafito, AM Shewan, W Yu, AN Combes, MH Little, ... Journal of Cell Biology 203 (2), 359-372 , 2013 2013 Citations: 69
Numerical study of polymer tumbling in linear shear flows A Puliafito, K Turitsyn Physica D: Nonlinear Phenomena 211 (1-2), 9-22 , 2005 2005 Citations: 67
The viscoelastic Kolmogorov flow: eddy viscosity and linear stability G Boffetta, A Celani, A Mazzino, A Puliafito, M Vergassola Journal of Fluid Mechanics 523, 161-170 , 2005 2005 Citations: 60
Live Monitoring of Blastemal Cell Contributions during Appendage Regeneration VA Tornini, A Puliafito, LA Slota, JD Thompson, G Nachtrab, AL Kaushik, ... Current Biology 26 (22), 2981-2991 , 2016 2016 Citations: 57
Three-dimensional chemotaxis-driven aggregation of tumor cells. A Puliafito, A De Simone, G Seano, PA Gagliardi, L Di Blasio, F Chianale, ... Scientific reports 5, 15205-15205 , 2015 2015 Citations: 56
PDK1-mediated activation of MRCKα regulates directional cell migration and lamellipodia retraction PA Gagliardi, L Di Blasio, A Puliafito, G Seano, R Sessa, F Chianale, ... Journal of Cell Biology 206 (3), 415-434 , 2014 2014 Citations: 51
