Krizia Sagini

Exercise-induced extracellular vesicles delay tumor development by igniting inflammation in an immunologically cold triple-negative breast cancer mouse model Mlynska A, Dobrovolskiene N, Suveizde K, Lukaseviciute G, Sagini K, Gracia BM, Romero S, Llorente A, Line A, Butkute A, Gudaite B, Venckunas T, Matuseviciene N, Pasukoniene V.J Sport Health Sci. 2025 Apr 16:101041. doi: 10.1016/j.jshs.2025.101041.

siRNA screening reveals that SNAP29 contributes to exosome release Hessvik NP, Sagini K, Romero S, Ramirez-Garrastacho M, Rodriguez M, Tutturen AEV, Kvalvaag A, Stang E, Brech A, Sandvig K, Llorente A.Cell Mol Life Sci. 2023 Jun 7;80(7):177. doi: 10.1007/s00018-023-04822-8.

Lipid Biomarkers in Liquid Biopsies: Novel Opportunities for Cancer Diagnosis Sagini K, Urbanelli L, Buratta S, Emiliani C, Llorente A.Pharmaceutics. 2023 Jan 28;15(2):437. doi: 10.3390/pharmaceutics15020437.

Drug-Induced Lysosomal Impairment Is Associated with the Release of Extracellular Vesicles Carrying Autophagy Markers Sagini K, Buratta S, Delo F, Pellegrino RM, Giovagnoli S, Urbanelli L, Emiliani C.Int J Mol Sci. 2021 Nov 29;22(23):12922. doi: 10.3390/ijms222312922.

An emerging focus on lipids in extracellular vesicles Skotland T, Sagini K, Sandvig K, Llorente A.Adv Drug Deliv Rev. 2020;159:308-321. doi: 10.1016/j.addr.2020.03.002.

Oncogenic H-Ras Expression Induces Fatty Acid Profile Changes in Human Fibroblasts and Extracellular Vesicles Sagini K, Urbanelli L, Costanzi E, Mitro N, Caruso D, Emiliani C, Buratta S.Int J Mol Sci. 2018 Nov 8;19(11):3515. doi: 10.3390/ijms19113515.

• Urbanelli, Lorena; Sagini, Krizia; Delo, Federica; Buratta, Sandra; Lucci, Jacopo & Mercati, Valentino [Show all 7 contributors for this article] (2025). Targeting pH Inversion in Prostate Cancer Cells: A Role for Systems of Molecules of Vegetal Origin. International Journal of Molecular Sciences. ISSN 1661-6596. 26(16). doi: 10.3390/ijms26167700. Full text in Research Archive Show summary

  Intracellular alkalosis and extracellular acidosis are two pathological features associated with malignant cells. They offer advantages in terms of invasiveness and proliferation. Extracellular acidification is the consequence of intracellular metabolic changes associated with a higher metabolic rate of cancer cells, potentially inducing dangerous intracellular acidification. To overcome this menace, malignant cells adapt themselves to export hydrogen ions. Therefore, it is reasonable that targeting intracellular alkalinization and extracellular acidification to prompt the reversal of such a pH gradient towards a condition comparable to normal, untransformed cells may represent a strategy helping to contrast malignant behavior. In the present study, we investigated in vitro, in prostate cancer cell models, the biological activity towards intracellular, extracellular and organelle pH of systems of molecules of vegetal origin. A few of these systems were shown to promote intracellular acidification in vitro, whereas others were shown to prevent extracellular acidification and promote lysosomal alkalinization in a cell type-dependent manner. This result clearly indicates that these systems may function as agents interfering with malignant cells inverted pH gradient. Further analysis would be necessary to unravel the cell type specificity of their effects, as well as their mechanism of action. Nevertheless, our proof-of-principle study provides evidence that such systems of molecules can be considered interesting agents in co-adjuvating anti-cancer therapies

• Sagini, Krizia; Selvarajah, Mughilan; Romero, Silvana Angelica Troncoso; Bassols, Eva Citores; Gracia, Beatriz Martín & Ramirez, Manuel [Show all 16 contributors for this article] (2025). The Redox Enzyme Thioredoxin Is Increased in Plasma Extracellular Vesicles From Endurance‐Trained Females in Response to Acute Exercise. The FASEB Journal. ISSN 0892-6638. 39(16). doi: 10.1096/fj.202500678r. Full text in Research Archive Show summary

  The effects of exercise are mediated by molecules secreted into the circulation in response to exercise. Extracellular vesicles (EVs) have recently appeared as possible vehicles for some of these molecules, and as mediators of tissue crosstalk during exercise. This study aimed to identify changes in the proteome of EVs released into circulation by endurance running‐trained females after a bout of 60‐min aerobic exercise consisting of incremental treadmill running. Blood samples were collected immediately before and after exercise, and EVs were separated from plasma and characterized by Western blot and electron microscopy. Mass spectrometry‐based proteomics analysis of EV preparations was then performed and showed that the levels of 9 proteins were differentially expressed in response to exercise. Thioredoxin, an enzyme that plays an essential role in the maintenance of redox homeostasis, showed the largest change. The increase in the levels of thioredoxin after exercise was also observed by Western blot analysis. Interestingly, post‐exercise plasma EVs were able to protect cells against oxidative stress. These results support the effect of physical exercise in modulating the levels of molecules involved in the maintenance of redox homeostasis, such as thioredoxin and other antioxidant enzymes, in plasma EVs. Although further studies are needed to fully understand the functional role of these molecules, it is intriguing to consider the protective health effects that exercise‐induced EVs may have against oxidative stressors.

• Mlynska, Agata; Dobrovolskiene, Neringa; Suveizde, Karolina; Lukaseviciute, Gabija; Sagini, Krizia & Martin-Gracia, Beatriz [Show all 14 contributors for this article] (2025). Exercise-induced extracellular vesicles delay tumor development by igniting inflammation in an immunologically cold triple-negative breast cancer mouse model. Journal of Sport and Health Science. ISSN 2095-2546. 14. doi: 10.1016/j.jshs.2025.101041. Full text in Research Archive Show summary

  Background Preclinical studies demonstrate that exercise reduces tumor incidence and growth. Rapid release of extracellular vesicles (EVs) during exercise suggests their potential role as mediators of exercise-induced systemic effects and physiological adaptation. This study investigated the impact of exercise-induced plasma EVs on tumor growth and immune tumor microenvironment in murine models of triple-negative breast cancer (TNBC): EO771 (a C57BL/6-derived TNBC cell line) and 4T1 (a BALB/c-derived TNBC cell line). Methods Size exclusion chromatography was used to isolate exercise-induced EVs from plasma of healthy female mice (BALB/c and C56BL/6, n = 30 per strain) that underwent ten 30-min moderate-intensity treadmill running sessions over 2 weeks. Nanoparticle tracking analysis, Western blot, and electron microscopy confirmed the presence of EVs in the samples. Tumor-bearing mice (n = 72 per strain) were administered with exercise-induced EVs before or/and after tumor implantation. Local and systemic immune responses were assessed using flow cytometry, enzyme-linked immunosorbent assay (ELISA), and quantitative polymerase chain reaction (qPCR). Results Administration of exercise-induced EVs, particularly before tumor implantation, significantly suppressed tumor growth and reduced tumor burden in both TNBC models. In EO771, endpoint tumor volumes were 278–330 mm³ in treated groups compared to 799 mm³ in untreated (p < 0.0001), while in 4T1, treated groups showed volumes of 287–564 mm³ vs. 696 mm³ in untreated (p = 0.0002). Notable differences in tumor-infiltrating lymphoid and myeloid cell subpopulations indicated immunomodulatory effects of exercise-induced EVs, particularly in the 4T1 model, where their continuous administration significantly increased intratumoral cluster of differentiation 8 (CD8) T lymphocyte proportion (5.77% vs. 0.90% in untreated, p < 0.0001). Similarly, in the EO771 model, exercise-induced EVs administered before tumor implantation led to a marked rise in intratumoral CD8 T lymphocytes (2.24% vs. 1.08% in untreated, p = 0.0181). Conclusion Our findings indicate that exercise-induced EV treatment elicits a pro-inflammatory antitumor immune response, suggesting a shift of immunologically cold TNBC tumors towards a more inflamed phenotype associated with better outcomes. Our study supports the further investigation of EVs as modulators of antitumor immunity and their potential utility in enhancing the efficacy of immunotherapy.

• Jayakumar, Nikhil; Villegas-Hernández, Luis Enrique; Zhao, Weisong; Mao, Hong; Dullo, Firehun Tsige & Tinguely, Jean-Claude [Show all 9 contributors for this article] (2025). Chip-based label-free incoherent super-resolution optical microscopy. Light: Science & Applications (LSA). ISSN 2095-5545. 14(1). doi: 10.1038/s41377-025-01914-x. Full text in Research Archive Show summary

  The photo-kinetics of fluorescent molecules have enabled the circumvention of the far-field optical diffraction limit. Despite its enormous potential, the necessity to label the sample may adversely influence the delicate biology under investigation. Thus, continued development efforts are needed to surpass the far-field label-free diffraction barrier. The statistical similarity or finite coherence of the scattered light off the sample in label-free mode hinders the application of existing super-resolution methods based on incoherent fluorescence imaging. In this article, we present physics and propose a methodology to circumvent this challenge by exploiting the photoluminescence (PL) of silicon nitride waveguides for near-field illumination of unlabeled samples. The technique is abbreviated EPSLON, Evanescently decaying Photoluminescence Scattering enables Label-free Optical Nanoscopy. We demonstrate that such an illumination has properties that mimic the photo-kinetics of nano-sized fluorescent molecules, i.e., such an illumination permits incoherence between the scattered fields from various locations on the sample plane. Thus, the illumination scheme enables the development of a far-field label-free incoherent imaging system that is linear in intensity and stable over time, thereby permitting the application of techniques like structured illumination microscopy (SIM) and intensity-fluctuation-based optical nanoscopy (IFON) in label-free mode to circumvent the diffraction limit. In this proof-of-concept work, we observed a two-point resolution of $$\sim$$ ~ 180 nm on super-resolved nanobeads and resolution improvements between 1.9× to 2.8× over the diffraction limit, as quantified using Fourier Ring Correlation (FRC), on various biological samples. We believe EPSLON is a step forward within the field of incoherent far-field label-free super-resolution microscopy that holds a key to investigating biological systems in their natural state without the need for exogenous labels.

• Zoofaghari, Mohammad; Sagini, Krizia; Damrath, Martin; Zargarnia, Azar; Flaten, Håkon & Veletic, Mladen [Show all 8 contributors for this article] (2025). In silico Study of Bloodstream Penetrating Extracellular Vesicles. IEEE Transactions on Molecular, Biological, and Multi-Scale Communications. 11(2), p. 166–175. doi: 10.1109/TMBMC.2025.3550323. Full text in Research Archive
• Llorente, Alicia; Brokāne, Agnese; Mlynska, Agata; Puurand, Marju; Sagini, Krizia & Folkmane, Signe [Show all 16 contributors for this article] (2024). From sweat to hope: The role of exercise-induced extracellular vesicles in cancer prevention and treatment. Journal of Extracellular Vesicles. 13(8). doi: 10.1002/jev2.12500. Full text in Research Archive Show summary

  The benefits of regular physical exercise on cancer prevention, as well as reducing fatigue, treatment side effects and recurrence, and improving quality of life and over-all survival of cancer patients, are increasingly recognised. Initial studies showed that the concentration of extracellular vesicles (EVs) increases during physical activity and that EVs carry biologically active cargo. These EVs are released by blood cells, skeletal muscle and other organs involved in exercise, thus suggesting that EVs may mediate tissue crosstalk during exercise. This possibility triggered a great interest in the study of the roles of EVs in systemic adaptation to exercise and in their potential applications in the prevention and treatment of various diseases, including cancer. This review presents studies exploring the concentration and molecular cargo of EVs released during exercise. Furthermore, we discuss putative stimuli that may trigger EV release from various cell types, the biological functions and the impact of exercise-induced EVs on cancer development and progression. Understanding the interplay between exercise, EVs, and cancer biology may offer insights into novel therapeutic strategies and preventive measures for cancer.

• Zoofaghari, Mohammad; Damrath, Martin; Veletic, Mladen; Sagini, Krizia; Bergsland, Jacob & Martinez, Alicia Llorente [Show all 7 contributors for this article] (2023). In Silico Model for Tumor Diagnosis based on Bloodstream Penetrating Extracellular Vesicles. In Eckford, Andrew & Noel, Adam (Ed.), The 10th Annual ACM International Conference on Nanoscale Computing and Communication. Association for Computing Machinery (ACM). ISSN 9798400700347. p. 129–135. doi: 10.1145/3576781.3608719. Full text in Research Archive Show summary

  During the last decade, extracellular vesicles (EVs) have emerged in the scientific community because of their potential as natural drug nanocarriers and biomarkers in disease diagnosis, especially for cancer. While a few studies have focused on the propagation and biodistribution of tumorderived EVs within the extracellular space or the bloodstream, there is a lack of simultaneous consideration of the transport of EVs within the blood vessels, surrounding tissue and tumor microenvironment. Here, we introduce an in silico model that simulates the release of EVs from cancer cells, their transport within the surrounding tissues, uptake into the bloodstream, and circulation within the segment of the blood vessels network comprising the arterial, capillary and venous vessels. By examining the obtained simulation results, we also propose a novel diagnostic procedure for sub-millimeter tumors based on the EV penetration rate into the circulatory system. Our simulations demonstrate the sensitivity of the approach to tumor size, whereas our findings further demonstrate that EVs offer a promising avenue for non-invasive disease diagnosis.

• Sagini, Krizia; Urbanelli, Lorena; Buratta, Sandra; Emiliani, Carla & Llorente, Alicia (2023). Lipid Biomarkers in Liquid Biopsies: Novel Opportunities for Cancer Diagnosis. Pharmaceutics. 15(2), p. 1–25. doi: 10.3390/pharmaceutics15020437. Full text in Research Archive Show summary

  Altered cellular metabolism is a well-established hallmark of cancer. Although most studies have focused on the metabolism of glucose and glutamine, the upregulation of lipid metabolism is also frequent in cells undergoing oncogenic transformation. In fact, cancer cells need to meet the enhanced demand of plasma membrane synthesis and energy production to support their proliferation. Moreover, lipids are precursors of signaling molecules, termed lipid mediators, which play a role in shaping the tumor microenvironment. Recent methodological advances in lipid analysis have prompted studies aimed at investigating the whole lipid content of a sample (lipidome) to unravel the complexity of lipid changes in cancer patient biofluids. This review focuses on the application of mass spectrometry-based lipidomics for the discovery of cancer biomarkers. Here, we have summarized the main lipid alteration in cancer patients’ biofluids and uncovered their potential use for the early detection of the disease and treatment selection. We also discuss the advantages of using biofluid-derived extracellular vesicles as a platform for lipid biomarker discovery. These vesicles have a molecular signature that is a fingerprint of their originating cells. Hence, the analysis of their molecular cargo has emerged as a promising strategy for the identification of sensitive and specific biomarkers compared to the analysis of the unprocessed biofluid.

• Hessvik, Nina Pettersen; Sagini, Krizia; Romero, Silvana Angelica Troncoso; Ramirez, Manuel; Rodriguez, Marta & Tutturen, Astrid Elisabeth Voorham [Show all 11 contributors for this article] (2023). siRNA screening reveals that SNAP29 contributes to exosome release. Cellular and Molecular Life Sciences (CMLS). ISSN 1420-682X. 80. doi: 10.1007/s00018-023-04822-8. Full text in Research Archive Show summary

  Cells release extracellular vesicles (EVs) of different sizes. Small EVs (< 200 nm) can originate from the fusion of multive- sicular bodies with the plasma membrane, i.e. exosomes, and from budding of the plasma membrane, i.e. small ectosomes. To investigate the molecular machinery required for the release of small EVs, we developed a sensitive assay based on incor- poration of radioactive cholesterol in EV membranes and used it in a siRNA screening. The screening showed that depletion of several SNARE proteins affected the release of small EVs. We focused on SNAP29, VAMP8, syntaxin 2, syntaxin 3 and syntaxin 18, the depletion of which reduced the release of small EVs. Importantly, this result was verified using gold standard techniques. SNAP29 depletion resulted in the largest effect and was further investigated. Immunoblotting analysis of small EVs showed that the release of several proteins considered to be associated with exosomes like syntenin, CD63 and Tsg101 was reduced, while the level of several proteins that have been shown to be released in ectosomes (annexins) or by secretory autophagy (LC3B and p62) was not affected by SNAP29 depletion. Moreover, these proteins appeared in different fractions when the EV samples were further separated by a density gradient. These results suggest that SNAP29 depletion mainly affects the secretion of exosomes. To investigate how SNAP29 affects exosome release, we used microscopy to study the distribution of MBVs using CD63 labelling and CD63-pHluorin to detect fusion events of MVBs with the plasma membrane. SNAP29 depletion caused a redistribution of CD63-labelled compartments but did not change the number of fusion events. Further experiments are therefore needed to fully understand the function of SNAP29. To conclude, we have developed a novel screening assay that has allowed us to identify several SNAREs involved in the release of small EVs.

• Nunzi, Emilia; Mezzasoma, Letizia; Bellezza, Ilaria; Zelante, Teresa; Orvietani, Pierluigi & Coata, Giuliana [Show all 15 contributors for this article] (2023). Microbiota-Associated HAF-EVs Regulate Monocytes by Triggering or Inhibiting Inflammasome Activation. International Journal of Molecular Sciences. ISSN 1661-6596. 24(3). doi: 10.3390/ijms24032527. Full text in Research Archive
• Zendrini, Andrea; Guerra, Giorgia; Sagini, Krizia; Vagner, Tatyana; Vizio, Dolores Di & Bergese, Paolo (2022). On the surface-to-bulk partition of proteins in extracellular vesicles. Colloids and Surfaces B: Biointerfaces. ISSN 0927-7765. 218. doi: 10.1016/j.colsurfb.2022.112728. Full text in Research Archive
• Mezzasoma, Letizia; Bellezza, Ilaria; Orvietani, Pierluigi; Manni, Giorgia; Gargaro, Marco & Sagini, Krizia [Show all 13 contributors for this article] (2022). Amniotic fluid stem cell-derived extracellular vesicles are independent metabolic units capable of modulating inflammasome activation in THP-1 cells. The FASEB Journal. ISSN 0892-6638. 36(4). doi: 10.1096/fj.202101657R. Full text in Research Archive
• Buratta, Sandra; Shimanaka, Y.; Costanzi, E.; Ni, S.; Urbanelli, Lorena & Kono, N. [Show all 13 contributors for this article] (2021). Lipotoxic stress alters the membrane lipid profile of extracellular vesicles released by Huh-7 hepatocarcinoma cells. Scientific Reports. 11(1). doi: 10.1038/s41598-021-84268-9. Full text in Research Archive
• Guerriero, Ilaria; Ramberg, Håkon Andre; Sagini, Krizia; Ramirez, Manuel; Tasken, Kristin Austlid & Llorente, Alicia (2021). Implication of β2-adrenergic receptor and miR-196a correlation in neurite outgrowth of LNCaP prostate cancer cells. PLOS ONE. 16:e025828(6), p. 1–11. doi: 10.1371/journal.pone.0253828. Full text in Research Archive Show summary

  The β2-adrenergic receptor has been shown to be involved in neuroendocrine differentiation and to contribute to the development of aggressive prostate cancer. In this study we have investigated whether miR-196a plays a role in the regulation of the β2-adrenergic receptor in the LNCaP prostate cancer cell line. Our results show that the expression of miR-196a is elevated in LNCaP prostate cancer cells with reduced levels of β2-adrenergic receptor after stably transfection with three different shRNAs. Furthermore, treatment with β-blockers showed that this upregulation is strictly related to the low levels of β2-adrenergic receptor and not to the inhibition of the receptor signaling activity. Finally, we found that the reduced ability of LNCaP cells with low levels of β2-adrenergic receptor to initiate neuroendocrine differentiation under androgen depletion conditions is mediated by miR-196a. In conclusion, this study provides the rational for a role of miR-196a in the β2-adrenergic receptor mediated neuroendocrine differentiation of LNCaP prostate cancer cells.

• Skotland, Tore; Sagini, Krizia; Sandvig, Kirsten & Llorente, Alicia (2020). An emerging focus on lipids in extracellular vesicles. Advanced Drug Delivery Reviews. ISSN 0169-409X. 159, p. 308–321. doi: 10.1016/j.addr.2020.03.002. Full text in Research Archive Show summary

  Extracellular vesicles contain a lipid bilayer membrane that protects the encapsulated material, such as proteins, nucleic acids, lipids and metabolites, from the extracellular environment. These vesicles are released from cells via different mechanisms. During recent years extracellular vesicles have been studied as possible biomarkers for different diseases, as biological nanoparticles for drug delivery, and in basic studies as a tool to understand the structure of biological membranes and the mechanisms involved in vesicular trafficking. Lipids are essential molecular components of extracellular vesicles, but at the moment our knowledge about the lipid composition and the function of lipids in these vesicles is limited. However, the interest of the research community in these molecules is increasing as their role in extracellular vesicles is starting to be acknowledged. In this review, we will present the status of the field and describe what is needed to bring it forward.

• Buratta, Sandra; Tancini, Brunella; Sagini, Krizia; Delo, Federica; Chiaradia, Elisabetta & Urbanelli, Lorena [Show all 7 contributors for this article] (2020). Lysosomal exocytosis, exosome release and secretory autophagy: The autophagic- and endo-lysosomal systems go extracellular. International Journal of Molecular Sciences. ISSN 1661-6596. 21(7). doi: 10.3390/ijms21072576. Full text in Research Archive
• Parfejevs, Vadims; Sagini, Krizia; Buss, Arturs; Sobolevska, Kristlne; Martinez, Alicia Llorente & Riekstiņa, Una [Show all 7 contributors for this article] (2020). Adult Stem Cell-Derived Extracellular Vesicles in Cancer Treatment: Opportunities and Challenges. Cells. 9(5), p. 1–26. doi: 10.3390/cells9051171. Full text in Research Archive

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• Telumyan, Elen; Gracia, Beatriz Martín; Sagini, Krizia; Andersen, Maria K.; Tessem, May-Britt & Llorente, Alicia [Show all 7 contributors for this article] (2026). NOR-EV annual meeting 2026. doi: https:/www.norev.org/. Full text in Research Archive
• Telumyan, Elen; Gracia, Beatriz Martín; Sagini, Krizia; Andersen, Maria K.; Tessem, May-Britt & Rye, Morten Beck [Show all 7 contributors for this article] (2025). Optimizing Protocols for Seminal Extracellular Vesicle Isolation in Prostate Cancer Research. Full text in Research Archive
• Telumyan, Elen; Gracia, Beatriz Martín; Sagini, Krizia; Andersen, Maria K.; Llorente, Alicia & Tessem, May-Britt [Show all 7 contributors for this article] (2024). Transcriptomic Biomarker for Prostate Cancer Active Surveillance in Extracellular Vesicles from Liquid Biopsies. Full text in Research Archive
• Telumyan, Elen; Gracia, Beatriz Martín; Sagini, Krizia; Andersen, Maria K.; Llorente, Alicia & Tessem, May-Britt [Show all 7 contributors for this article] (2024). A Transcriptomic Biomarker for Prostate Cancer Active Surveillance in Extracellular Vesicles from Liquid Biopsies. Full text in Research Archive
• Telumyan, Elen; Gracia, Beatriz Martín; Sagini, Krizia; Andersen, Maria K.; Llorente, Alicia & Tessem, May-Britt [Show all 7 contributors for this article] (2024). Transcriptomic Biomarker for Prostate Cancer Active Surveillance in Extracellular Vesicles from Liquid Biopsies. Full text in Research Archive

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