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1. Results of a prospective observational study of autologous peripheral blood mononuclear cell therapy for no-option critical limb-threatening ischemia and severe diabetic foot ulcers

Author

Panunzi, Andrea, Madotto, Fabiana, Sangalli, Elena, Riccio, Federica, Sganzaroli, Adriana Barbara, Galenda, Paolo, Bertulessi, Amelia, Barmina, Maria Francesca, Ludovico, Ornella, Fortunato, Orazio, Setacci, Francesco, Airoldi, Flavio, Tavano, Davide, Giurato, Laura, Meloni, Marco, Uccioli, Luigi, Bruno, Antonino, Spinetti, Gaia, and Caravaggi, Carlo Maria Ferdinando

Published
2022
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4. The longevity-associated BPIFB4 gene supports cardiac function and vascularization in ageing cardiomyopathy

Author

Cattaneo, Monica, primary, Beltrami, Antonio P, additional, Thomas, Anita C, additional, Spinetti, Gaia, additional, Alvino, Valeria Vincenza, additional, Avolio, Elisa, additional, Veneziano, Claudia, additional, Rolle, Irene Giulia, additional, Sponga, Sandro, additional, Sangalli, Elena, additional, Maciag, Anna, additional, Dal Piaz, Fabrizio, additional, Vecchione, Carmine, additional, Alenezi, Aishah, additional, Paisey, Stephen, additional, Puca, Annibale A, additional, and Madeddu, Paolo, additional

Published
2023
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6. Sensory neuropathy hampers nociception-mediated bone marrow stem cell release in mice and patients with diabetes

Author

Dang, Zexu, Maselli, Davide, Spinetti, Gaia, Sangalli, Elena, Carnelli, Franco, Rosa, Francesco, Seganfreddo, Elena, Canal, Fabio, Furlan, Anna, Paccagnella, Agostino, Paiola, Emanuela, Lorusso, Bruno, Specchia, Claudia, Albiero, Mattia, Cappellari, Roberta, Avogaro, Angelo, Falco, Angela, Quaini, Federico, Ou, Kepeng, Rodriguez-Arabaolaza, Iker, Emanueli, Costanza, Sambataro, Maria, Fadini, Gian Paolo, and Madeddu, Paolo

Published
2015
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7. Additional file 1 of Results of a prospective observational study of autologous peripheral blood mononuclear cell therapy for no-option critical limb-threatening ischemia and severe diabetic foot ulcers

Author

Panunzi, Andrea, Madotto, Fabiana, Sangalli, Elena, Riccio, Federica, Sganzaroli, Adriana Barbara, Galenda, Paolo, Bertulessi, Amelia, Barmina, Maria Francesca, Ludovico, Ornella, Fortunato, Orazio, Setacci, Francesco, Airoldi, Flavio, Tavano, Davide, Giurato, Laura, Meloni, Marco, Uccioli, Luigi, Bruno, Antonino, Spinetti, Gaia, and Caravaggi, Carlo Maria Ferdinando

Abstract

Additional file 1: Figure S1 Panel A-B. Representative case of gangrene of the big toe. Panel C. PB-MNCs injection procedure was performed along the relevant axis below the knee, below the ankle, and closed to the wound area, at intervals of 1–2 cm and to a mean depth of 1.5–2 cm, using a 21G needle.Panel D. Patient submitted to big toe necrosectomy with bone exposure and dressing with Bioinductive Dermal Substitute based on Hyaluronic Acid. Figure S2 Representative dot plots for the gating strategy to detect the relative frequency (expressed as % of cells positive for the selected surface antigens and their combination) of the following PB-MNCs populations, as determined by multicolor flow cytometry. For rare events (CD34+ and CD34+CXCR4+ cells), the large dot option, by FlowJo software, was used. FSC: forward scatter, SSC: side scatter. Figure S3 Representative images of two clinical cases. Panel A. Representative pictures of a clinical case of PB-MNCs therapy and right Chopart Amputation. From left to right: (i) before PB-MNCs therapy; (ii) at the time of the 1st treatment; (iii) at the 2nd treatment; (iv) at the 3rd treatment; (v) ulcer healing; (vi) definitive orthosis employing Below the Knee rigid casting with semi-rigid sole with high energy release to allow correct residual walking capacity. Panel B. Forefoot-mid foot gangrene in severe CLTI with no distal vessels run-off submitted to PB-MNC therapy. (i) Dorsal and (ii) plantar aspect of the foot; (iii) the aspect of the foot with secondary bone coverage and new dermal tissues coming from dermal substitute HA-based dressing. (iv) Dorsal and (v) plantar aspect of the foot at the indicated time after treatment. (vi) Correct orthesis for transmetatarsal amputation: custom-made shoes with lateral and medial reinforcement, biomechanics rigid rocker bottom sole, and fitting multilayer insole. Figure S4 Probability of ulcer healing without a major amputation during the 1st year after treatment in the study population stratified by TcPO2 at the last treatment (panel A); during the whole follow-up period in the study population stratified by TcPO2 at the last treatment (panel B). Patients who died or drop out were not counted as “at-risk” (censored subjects). Figure S5 Relative abundance of CD34+ subpopulations during PB-MNCs therapy stratified by ulcer healing without a major amputation after 1 year. Panel A. CD34+. Panel B. CD34+CXCR4+ HSPCs. The grey box represents patients with healing and the white box represents patients without healing at 1-year follow-up. No statistical differences were observed. Table S1 Perfusion and cellular characterization at enrollement, stratifying study population according to ulcer healing without a major amputation at 1 year. Table S2 Clinical outcomes occurred during follow-up, stratifying the study population according to TcPO2 levels after the last PB-MNCs treatment (< 40 mmHg; ≥ 40 mmHg).

Published
2022
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8. Hematopoietic progenitor cell liabilities and alarmins S100A8/A9‐related inflammaging associate with frailty and predict poor cardiovascular outcomes in older adults

Author

Bonora, Benedetta Maria, primary, Palano, Maria Teresa, additional, Testa, Gianluca, additional, Fadini, Gian Paolo, additional, Sangalli, Elena, additional, Madotto, Fabiana, additional, Persico, Giuseppe, additional, Casciaro, Francesca, additional, Vono, Rosa, additional, Colpani, Ornella, additional, Scavello, Francesco, additional, Cappellari, Roberta, additional, Abete, Pasquale, additional, Orlando, Patrizia, additional, Carnelli, Franco, additional, Berardi, Andrea Giovanni, additional, De Servi, Stefano, additional, Raucci, Angela, additional, Giorgio, Marco, additional, Madeddu, Paolo, additional, and Spinetti, Gaia, additional

Published
2022
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9. Abstract P452: Stem Cells Defect And Alarmins S100A8/A9 Overexpression Associate With Early-stage Frailty And Poor Cardiovascular Health In Older Adults

Author

Bonora, Benedetta Maria, primary, Palano, Maria Teresa, additional, Testa, Gianluca, additional, Fadini, Gian Paolo, additional, Sangalli, Elena, additional, Madotto, Fabiana, additional, Persico, Giuseppe, additional, Casciaro, Francesca, additional, Vono, Rosa, additional, Colpani, Ornella, additional, Scavello, Francesco, additional, Cappellari, Roberta, additional, Orlando, Patrizia, additional, Carnelli, Franco, additional, Berardi, Andrea, additional, De Servi, Stefano, additional, Raucci, Angela, additional, ABETE, PASQUALE, additional, Giorgio, Marco, additional, Madeddu, Paolo, additional, and Spinetti, Gaia, additional

Published
2021
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14. MicroRNA-21/PDCD4 proapoptotic signaling from circulating CD34+ cells to vascular endothelial cells: a potential contributor to adverse cardiovascular outcomes in patients with critical limb ischemia

Author

Spinetti, Gaia, primary, Sangalli, Elena, primary, Tagliabue, Elena, primary, Maselli, Davide, primary, Colpani, Ornella, primary, Ferland-McCollough, David, primary, Carnelli, Franco, primary, Orlando, Patrizia, primary, Paccagnella, Agostino, primary, Furlan, Anna, primary, Stefani, Piero Maria, primary, Sambado, Luisa, primary, Sambataro, Maria, primary, and Madeddu, Paolo, primary

Published
2020
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15. Circulating MicroRNA-15a Associates With Retinal Damage in Patients With Early Stage Type 2 Diabetes

Author

Sangalli, Elena, primary, Tagliabue, Elena, additional, Sala, Lucia La, additional, Prattichizzo, Francesco, additional, Uccellatore, AnnaChiara, additional, Spada, Daniela, additional, Lorino, Fabrizio, additional, Candia, Paola de, additional, Lupini, Silvia, additional, Cantone, Laura, additional, Favero, Chiara, additional, Madeddu, Paolo, additional, Bollati, Valentina, additional, Genovese, Stefano, additional, and Spinetti, Gaia, additional

Published
2020
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16. Circulating microRNA-21 is an early predictor of ROS-mediated damage in subjects with high risk of developing diabetes and in drug-naïve T2D 11 Medical and Health Sciences 1103 Clinical Sciences

Author

La Sala, Lucia, Mrakic-Sposta, Simona, Tagliabue, Elena, Prattichizzo, Francesco, Micheloni, Stefano, Sangalli, Elena, Specchia, Claudia, Uccellatore, Anna Chiara, Lupini, Silvia, Spinetti, Gaia, De Candia, Paola, and Ceriello, Antonio

Subjects
Diabetes and Metabolism, Endocrinology, Antioxidant response, Diabetes, Internal Medicine, IGT, T2D, miR-21, Prediabetes, ROS homeostasis, SOD2, Endocrinology, Diabetes and Metabolism, Cardiology and Cardiovascular Medicine
Published
2019

17. MOESM1 of Circulating microRNA-21 is an early predictor of ROS-mediated damage in subjects with high risk of developing diabetes and in drug-naïve T2D

Author

Sala, Lucia La, Mrakic-Sposta, Simona, Tagliabue, Elena, Prattichizzo, Francesco, Micheloni, Stefano, Sangalli, Elena, Specchia, Claudia, Uccellatore, Anna, Lupini, Silvia, Spinetti, Gaia, Candia, Paola De, and Ceriello, Antonio

Abstract

Additional file 1. Absorbance (Abs) of plasma samples read at 375 nm and 414 nm wavelength using a spectrophotometer [26, 27]. The ratio between absorbance at 414 nm and 375 nm was calculated. The ratio major than 1.4 was considered sample hemolyzed [26]. Further, we read at 541 nm and 576 nm in order to verify the high levels of hemolysis [26, 27].

Published
2019
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18. Corrigendum: 3D hydrogel environment rejuvenates aged pericytes for skeletal muscle tissue engineering

Author

Fuoco, Claudia, primary, Sangalli, Elena, additional, Vono, Rosa, additional, Testa, Stefano, additional, Sacchetti, Benedetto, additional, Latronico, Michael V. G., additional, Bernardini, Sergio, additional, Madeddu, Paolo, additional, Cesareni, Gianni, additional, Seliktar, Dror, additional, Rizzi, Roberto, additional, Bearzi, Claudia, additional, Cannata, Stefano M., additional, Spinetti, Gaia, additional, and Gargioli, Cesare, additional

Published
2019
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20. MicroRNA-21/PDCD4 Proapoptotic Signaling From Circulating CD34+ Cells to Vascular Endothelial Cells: A Potential Contributor to Adverse Cardiovascular Outcomes in Patients With Critical Limb Ischemia.

Author

Spinetti, Gaia, Sangalli, Elena, Tagliabue, Elena, Maselli, Davide, Colpani, Ornella, Ferland-McCollough, David, Carnelli, Franco, Orlando, Patrizia, Paccagnella, Agostino, Furlan, Anna, Stefani, Piero Maria, Sambado, Luisa, Sambataro, Maria, and Madeddu, Paolo

Subjects
*VASCULAR endothelial cells, *BONE marrow cells, *CELL migration, *ENDOTHELIAL cells, *TYPE 2 diabetes
Abstract

OBJECTIVE: In patients with type 2 diabetes (T2D) and critical limb ischemia (CLI), migration of circulating CD34+ cells predicted cardiovascular mortality at 18 months after revascularization. This study aimed to provide long-term validation and mechanistic understanding of the biomarker. RESEARCH DESIGN AND METHODS: The association between CD34+ cell migration and cardiovascular mortality was reassessed at 6 years after revascularization. In a new series of T2D-CLI and control subjects, immuno-sorted bone marrow CD34+ cells were profiled for miRNA expression and assessed for apoptosis and angiogenesis activity. The differentially regulated miRNA-21 and its proapoptotic target, PDCD4, were titrated to verify their contribution in transferring damaging signals from CD34+ cells to endothelial cells. RESULTS: Multivariable regression analysis confirmed that CD34+ cell migration forecasts long-term cardiovascular mortality. CD34+ cells from T2D-CLI patients were more apoptotic and less proangiogenic than those from control subjects and featured miRNA-21 downregulation, modulation of several long noncoding RNAs acting as miRNA-21 sponges, and upregulation of the miRNA-21 proapoptotic target PDCD4. Silencing miR-21 in control CD34+ cells phenocopied the T2D-CLI cell behavior. In coculture, T2D-CLI CD34+ cells imprinted naive endothelial cells, increasing apoptosis, reducing network formation, and modulating the TUG1 sponge/miRNA-21/PDCD4 axis. Silencing PDCD4 or scavenging reactive oxygen species protected endothelial cells from the negative influence of T2D-CLI CD34+ cells. CONCLUSIONS: Migration of CD34+ cells predicts long-term cardiovascular mortality in T2D-CLI patients. An altered paracrine signaling conveys antiangiogenic and proapoptotic features from CD34+ cells to the endothelium. This damaging interaction may increase the risk for life-threatening complications. [ABSTRACT FROM AUTHOR]

Published
2020
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21. Activation of the Pro-Oxidant PKCβII-p66Shc Signaling Pathway Contributes to Pericyte Dysfunction in Skeletal Muscles of Patients With Diabetes With Critical Limb Ischemia

Author

Vono, Rosa, Fuoco, Claudia, Testa, Stefano, Pirro, Stefano, Maselli, David, Ferl, David, McCollough, Sangalli, Elena, Pintus, Gianfranco, Giordo, Roberta, Finzi, Giovanna, Sessa, Fausto, Cardani, Rosanna, Gotti, Ambra, Losa, Sergio, Cesareni, Gianni, Rizzi, Roberto, Bearzi, Claudia, Cannata, Stefano, Spinetti, Gaia, Gargioli, Cesare, and Madeddu, Paolo

Subjects
Male, 0301 basic medicine, Endocrinology, Diabetes and Metabolism, medicine.disease_cause, Endocrinology, Ischemia, Myocyte, Medicine, Endocrinology, Diabetes, Microscopy, Blotting, Myogenesis, Settore BIO/13, Diabetes, Skeletal, Flow Cytometry, Immunohistochemistry, 3. Good health, medicine.anatomical_structure, Limb Ischemia, Muscle, Female, Pericyte, medicine.symptom, Western, Signal Transduction, medicine.medical_specialty, Blotting, Western, Enzyme-Linked Immunosorbent Assay, Phthalimides, In Vitro Techniques, Electron, 03 medical and health sciences, Microscopy, Electron, Transmission, Downregulation and upregulation, Diabetic Patients, Diabetes mellitus, Internal medicine, Protein Kinase C beta, Human Umbilical Vein Endothelial Cells, Internal Medicine, medicine, Transmission, Humans, Muscle, Skeletal, Aged, Cell Proliferation, business.industry, Critical limb ischemia, medicine.disease, Oxidative Stress, Pericytes, Reactive Oxygen Species, Surgery, 030104 developmental biology, business, skeletal muscles, Oxidative stress
Abstract

Critical limb ischemia (CLI), foot ulcers, former amputation, and impaired regeneration are independent risk factors for limb amputation in subjects with diabetes. The present work investigates whether and by which mechanism diabetes negatively impacts on functional properties of muscular pericytes (MPs), which are resident stem cells committed to reparative angiomyogenesis. We obtained muscle biopsy samples from patients with diabetes who were undergoing major limb amputation and control subjects. Diabetic muscles collected at the rim of normal tissue surrounding the plane of dissection showed myofiber degeneration, fat deposition, and reduction of MP vascular coverage. Diabetic MPs (D-MPs) display ultrastructural alterations, a differentiation bias toward adipogenesis at the detriment of myogenesis and an inhibitory activity on angiogenesis. Furthermore, they have an imbalanced redox state, with downregulation of the antioxidant enzymes superoxide dismutase 1 and catalase, and activation of the pro-oxidant protein kinase C isoform -II (PKCII)- dependent p66Shc signaling pathway. A reactive oxygen species scavenger or, even more effectively, clinically approved PKCII inhibitors restore D-MP angiomyogenic activity. Inhibition of the PKCII-dependent p66Shc signaling pathway could represent a novel therapeutic approach for the promotion of muscle repair in individuals with diabetes. This work has been supported by British Heart Foundation grants RJ5905 and RM/13/2/30158 and Italian Ministry of Health grant RF-2011-02346867 to P.M., Cariplo Foundation grant 2013-0887 to G.S., European Research Council grant N322749 DEPTH to G.C., and Uncovering Excellence Grant 2014 MDESMPLAT to C.G. Scopus

Published
2016
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22. Central role of the p53 pathway in the noncoding-RNA response to oxidative stress

Author

Fuschi, Paola, primary, Carrara, Matteo, additional, Voellenkle, Christine, additional, Garcia-Manteiga, Jose Manuel, additional, Righini, Paolo, additional, Maimone, Biagina, additional, Sangalli, Elena, additional, Villa, Francesco, additional, Specchia, Claudia, additional, Picozza, Mario, additional, Nano, Giovanni, additional, Gaetano, Carlo, additional, Spinetti, Gaia, additional, Puca, Annibale A., additional, Magenta, Alessandra, additional, and Martelli, Fabio, additional

Published
2017
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23. A unique plasma microRNA profile defines type 2 diabetes progression

Author

de Candia, Paola, primary, Spinetti, Gaia, additional, Specchia, Claudia, additional, Sangalli, Elena, additional, La Sala, Lucia, additional, Uccellatore, Annachiara, additional, Lupini, Silvia, additional, Genovese, Stefano, additional, Matarese, Giuseppe, additional, and Ceriello, Antonio, additional

Published
2017
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24. Central role of the p53 pathway in the noncoding-RNA response to oxidative stress

Author

Fuschi, Paola, Carrara, Matteo, Voellenkle, Christine, Garcia‐Manteiga, Jose Manuel, Righini, Paolo, Maimone, Biagina, Sangalli, Elena, Villa, Francesco, Specchia, Claudia, Picozza, Mario, Nano, Giovanni, Gaetano, Carlo, Spinetti, Gaia, Puca, Annibale A., Magenta, Alessandra, Martelli, Fabio, Fuschi, Paola, Carrara, Matteo, Voellenkle, Christine, Garcia‐Manteiga, Jose Manuel, Righini, Paolo, Maimone, Biagina, Sangalli, Elena, Villa, Francesco, Specchia, Claudia, Picozza, Mario, Nano, Giovanni, Gaetano, Carlo, Spinetti, Gaia, Puca, Annibale A., Magenta, Alessandra, and Martelli, Fabio

Abstract

Oxidative stress plays a fundamental role in many conditions. Specifically, redox imbalance inhibits endothelial cell (EC) growth, inducing cell death and senescence. We used global transcriptome profiling to investigate the involvement of noncoding-RNAs in these phenotypes. By RNA-sequencing, transcriptome changes were analyzed in human ECs exposed to H2O2, highlighting a pivotal role of p53-signaling. Bioinformatic analysis and validation in p53-silenced ECs, identified several p53-targets among both mRNAs and long noncoding-RNAs (lncRNAs), including MALAT1 and NEAT1. Among microRNAs (miRNAs), miR-192-5p was the most induced by H2O2 treatment, in a p53-dependent manner. Down-modulated mRNA-targets of miR-192-5p were involved in cell cycle, DNA repair and stress response. Accordingly, miR-192-5p overexpression significantly decreased EC proliferation, inducing cell death. A central role of the p53-pathway was also confirmed by the analysis of differential exon usage: Upon H2O2 treatment, the expression of p53-dependent 5’-isoforms of MDM2 and PVT1 increased selectively. The transcriptomic alterations identified in H2O2-treated ECs were also observed in other physiological and pathological conditions where redox control plays a fundamental role, such as ECs undergoing replicative senescence, skeletal muscles of critical limb-ischemia patients and the peripheral-blood mononuclear cells of long-living individuals. Collectively, these findings indicate a prominent role of noncoding-RNAs in oxidative stress response.

Published
2017

25. The expression of the BPIFB4 and CXCR4 associates with sustained health in long-living individuals from Cilento-Italy

Author

Spinetti, Gaia, primary, Sangalli, Elena, additional, Specchia, Claudia, additional, Villa, Francesco, additional, Spinelli, Chiara, additional, Pipolo, Rita, additional, Carrizzo, Albino, additional, Greco, Simona, additional, Voellenkle, Christine, additional, Vecchione, Carmine, additional, Madeddu, Paolo, additional, Martelli, Fabio, additional, and Puca, Annibale Alessandro, additional

Published
2017
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26. Activation of the Pro-Oxidant PKCβII-p66Shc Signaling Pathway Contributes to Pericyte Dysfunction in Skeletal Muscles of Patients With Diabetes With Critical Limb Ischemia

Author

Vono, Rosa, primary, Fuoco, Claudia, additional, Testa, Stefano, additional, Pirrò, Stefano, additional, Maselli, Davide, additional, Ferland McCollough, David, additional, Sangalli, Elena, additional, Pintus, Gianfranco, additional, Giordo, Roberta, additional, Finzi, Giovanna, additional, Sessa, Fausto, additional, Cardani, Rosanna, additional, Gotti, Ambra, additional, Losa, Sergio, additional, Cesareni, Gianni, additional, Rizzi, Roberto, additional, Bearzi, Claudia, additional, Cannata, Stefano, additional, Spinetti, Gaia, additional, Gargioli, Cesare, additional, and Madeddu, Paolo, additional

Published
2016
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28. Genetic Analysis Reveals a Longevity-Associated Protein Modulating Endothelial Function and Angiogenesis

Author

Villa, Francesco, primary, Carrizzo, Albino, additional, Spinelli, Chiara C., additional, Ferrario, Anna, additional, Malovini, Alberto, additional, Maciąg, Anna, additional, Damato, Antonio, additional, Auricchio, Alberto, additional, Spinetti, Gaia, additional, Sangalli, Elena, additional, Dang, Zexu, additional, Madonna, Michele, additional, Ambrosio, Mariateresa, additional, Sitia, Leopoldo, additional, Bigini, Paolo, additional, Calì, Gaetano, additional, Schreiber, Stefan, additional, Perls, Thomas, additional, Fucile, Sergio, additional, Mulas, Francesca, additional, Nebel, Almut, additional, Bellazzi, Riccardo, additional, Madeddu, Paolo, additional, Vecchione, Carmine, additional, and Puca, Annibale A., additional

Published
2015
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29. Increased Antioxidant Defense Mechanism in Human Adventitia-Derived Progenitor Cells Is Associated with Therapeutic Benefit in Ischemia

Author

Iacobazzi, Dominga, primary, Mangialardi, Giuseppe, additional, Gubernator, Miriam, additional, Hofner, Manuela, additional, Wielscher, Matthias, additional, Vierlinger, Klemens, additional, Reni, Carlotta, additional, Oikawa, Atsuhiko, additional, Spinetti, Gaia, additional, Vono, Rosa, additional, Sangalli, Elena, additional, Montagnani, Monica, additional, and Madeddu, Paolo, additional

Published
2014
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30. 3D hydrogel environment rejuvenates aged pericytes for skeletal muscle tissue engineering

Author

Fuoco, Claudia, primary, Sangalli, Elena, additional, Vono, Rosa, additional, Testa, Stefano, additional, Sacchetti, Benedetto, additional, Latronico, Michael V. G., additional, Bernardini, Sergio, additional, Madeddu, Paolo, additional, Cesareni, Gianni, additional, Seliktar, Dror, additional, Rizzi, Roberto, additional, Bearzi, Claudia, additional, Cannata, Stefano M., additional, Spinetti, Gaia, additional, and Gargioli, Cesare, additional

Published
2014
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31. Global Remodeling of the Vascular Stem Cell Niche in Bone Marrow of Diabetic Patients

Author

Spinetti, Gaia, primary, Cordella, Daniela, additional, Fortunato, Orazio, additional, Sangalli, Elena, additional, Losa, Sergio, additional, Gotti, Ambra, additional, Carnelli, Franco, additional, Rosa, Francesco, additional, Riboldi, Stefano, additional, Sessa, Fausto, additional, Avolio, Elisa, additional, Beltrami, Antonio Paolo, additional, Emanueli, Constanza, additional, and Madeddu, Paolo, additional

Published
2013
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32. MicroRNA-15a and MicroRNA-16 Impair Human Circulating Proangiogenic Cell Functions and Are Increased in the Proangiogenic Cells and Serum of Patients With Critical Limb Ischemia

Author

Spinetti, Gaia, primary, Fortunato, Orazio, additional, Caporali, Andrea, additional, Shantikumar, Saran, additional, Marchetti, Micol, additional, Meloni, Marco, additional, Descamps, Betty, additional, Floris, Ilaria, additional, Sangalli, Elena, additional, Vono, Rosa, additional, Faglia, Ezio, additional, Specchia, Claudia, additional, Pintus, Gianfranco, additional, Madeddu, Paolo, additional, and Emanueli, Costanza, additional

Published
2013
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33. Abstract 14793: Molecular Transferring of Anti-Angiogenic and Pro-Apoptotic Signaling From Diabetic CD34+ Cells to the Vascular Endothelium via Activation of the microRNA-21/PDCD4 Pathway

Author

Spinetti, Gaia, Sangalli, Elena, Maselli, Davide, Vono, Rosa, Colpani, Ornella, Tagliabue, Elena, Ferland-McCollough, David, Carnelli, Franco, Orlando, Patrizia, Sambado, Luisa, Sambataro, Maria, and Madeddu, Paolo R

Abstract

Introduction:We recently demonstrated that enhanced in-vitro migratory ability of CD45dimKDR+CXCR4+ CD34+ cells predicts cardiovascular mortality 18 months after percutaneous angioplasty in people with type 2 diabetes mellitus (DM) and critical limb ischemia (CLI).Hypothesis:We hypothesize that DM+CLI convert migratory proangiogenic CD34+ cells to negative regulators of endothelial function, thus predisposing to cardiovascular events.Methods:We analyzed the rate of mortality in the previously studied cohort of 119 subjects at a longer follow up (six-year). In addition, we compared apoptosis and angiogenic capability of MAC-sorted BM-CD34+ cells in newly-recruited subjects with DM+CLI and control without DM (ctr). Following microRNA (miR) profiling (N=6 donor/group), we validated modulated miRs in new donors BM-CD34+ cells. Finally, we focused on miR-21 and its target, PDCD4, an apoptosis regulator on functional outcomes.Results:The percentage of migrated CD45dimCD34+CXCR4+KDR+ cells was significantly higher in subjects who died for CV events compared with event-free subjects (0.05 ? 0.08 vs 0.02 ? 0.03; p-value=0.0372). Cell migration was associated with an increased risk of CV death in a multivariable model adjusted by age and coronary artery disease (p-value=0.0007 and 0.0124, respectively). In vitro studies demonstrated that DM+CLI-BM-CD34+ cells undergo apoptosis and are less proangiogenic than ctr-cells. We observed down-modulation of miR-21 and reciprocal increase in its target, PDCD4, in DM+CLI-CD34+ cells. Silencing miR-21 in ctr-CD34+ cells phenocopied DM+CLI behavior. Likewise, exposure of endothelial cells to patient-derived CD34+ cells induced apoptosis and reduced angiogenesis in association with miR-21/PDCD4 modulation. Silencing endothelial PDCD4 and treatment with ROS-scavenger NAC reversed these effects.Conclusions:Increased migratory activity of CD34+ cells associates with long-term cardiovascular mortality in complicated diabetic patients. An altered paracrine ROS/miR-21/PDCD4 pathway may transfer a damging signaling from diabetic CD34+ cells to the vascular endothelium, thereby increasing the risk for adverse cardiovascular complications and death.

Published
2019
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34. Circulating MicroRNA-15a Associates With Retinal Damage in Patients With Early Stage Type 2 Diabetes

Author

Elena Sangalli, Chiara Favero, Elena Tagliabue, Laura Cantone, Paola de Candia, Lucia La Sala, Silvia Lupini, Anna Chiara Uccellatore, Gaia Spinetti, Stefano Genovese, Francesco Prattichizzo, Fabrizio Lorino, Paolo Madeddu, Daniela Spada, Valentina Bollati, Sangalli, Elena, Tagliabue, Elena, Sala, Lucia La, Prattichizzo, Francesco, Uccellatore, Annachiara, Spada, Daniela, Lorino, Fabrizio, de Candia, Paola, Lupini, Silvia, Cantone, Laura, Favero, Chiara, Madeddu, Paolo, Bollati, Valentina, Genovese, Stefano, and Spinetti, Gaia

Subjects
0301 basic medicine, Retinal degeneration, Male, medicine.medical_specialty, endocrine system diseases, Endocrinology, Diabetes and Metabolism, 030209 endocrinology & metabolism, Type 2 diabetes, Gastroenterology, lcsh:Diseases of the endocrine glands. Clinical endocrinology, prediction and prevention of type 2 diabetes, Impaired glucose tolerance, Cohort Studies, 03 medical and health sciences, 0302 clinical medicine, Endocrinology, Retinal Diseases, Internal medicine, Diabetes mellitus, retinopathy, medicine, Humans, Circulating MicroRNA, Original Research, Glucose tolerance test, lcsh:RC648-665, medicine.diagnostic_test, business.industry, Type 2 Diabetes Mellitus, nutritional and metabolic diseases, Middle Aged, medicine.disease, Prognosis, microvascular disease, MicroRNAs, 030104 developmental biology, microRNA-15a, Diabetes Mellitus, Type 2, Female, extracellular vesicle, business, extracellular vesicles, Body mass index, Biomarkers, prediction and prevention of type 2 diabete, Retinopathy, Follow-Up Studies
Abstract

Circulating microRNAs are potential biomarkers of type 2 diabetes mellitus (T2DM) and related complications. Here, we investigated the association of microRNA-15a with early retinal damage in T2DM. A cohort of untreated subjects screened for intermediate/high risk of T2DM, according to a score assessment questionnaire, and then recognized to have a normal (NGT) or impaired (IGT) glucose tolerance or T2DM was studied. The thickness of the ganglion cell complex (GCC), an early marker of retinal degeneration anteceding overt retinopathy was assessed by Optical Coherence Tomography. Total and extracellular vesicles (EV)-associated microRNA-15a quantity was measured in plasma by real time PCR. MicroRNA-15a level was significantly higher in subjects with IGT and T2DM compared with NGT. MicroRNA-15a abundance was correlated to body mass index and classical diabetes biomarkers, including fasting glucose, HbA1c, insulinemia, and HOMA-IR. Moreover, GCC thickness was significantly reduced in IGT and T2DM subjects compared with NGT controls. Importantly, total microRNA-15a correlated with GCC in IGT subjects, while in T2DM subjects, EV-microRNA-15a negatively correlated with GCC, suggesting that microRNA-15a may monitor initial retinal damage. The assessment of plasma microRNA-15a may help refining risk assessment and secondary prevention in patients with preclinical T2DM.

Published
2020
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35. A unique plasma microRNA profile defines type 2 diabetes progression

Author

Lucia La Sala, Antonio Ceriello, Stefano Genovese, Gaia Spinetti, Giuseppe Matarese, Elena Sangalli, Claudia Specchia, Annachiara Uccellatore, Silvia Lupini, Paola de Candia, De Candia, Paola, Spinetti, Gaia, Specchia, Claudia, Sangalli, Elena, Sala, Lucia La, Uccellatore, Annachiara, Lupini, Silvia, Genovese, Stefano, Matarese, Giuseppe, and Ceriello, Antonio

Subjects
0301 basic medicine, Blood Glucose, Male, Genetics and Molecular Biology (all), endocrine system diseases, Physiology, lcsh:Medicine, Type 2 diabetes, Biochemistry, Impaired glucose tolerance, chemistry.chemical_compound, Endocrinology, 0302 clinical medicine, Glucose Metabolism, Risk Factors, Blood plasma, Medicine and Health Sciences, Diabetes diagnosis and management, lcsh:Science, Glucose Tolerance, Glucose tolerance test, Multidisciplinary, medicine.diagnostic_test, Middle Aged, Type 2 Diabetes, Body Fluids, 3. Good health, Nucleic acids, Blood, Cholesterol, Disease Progression, Carbohydrate Metabolism, Female, Anatomy, Aged, Biomarkers, Diabetes Mellitus, Type 2, Glucose Intolerance, Glucose Tolerance Test, Humans, MicroRNAs, Prediabetic State, Retrospective Studies, Biochemistry, Genetics and Molecular Biology (all), Agricultural and Biological Sciences (all), Type 2, Research Article, medicine.medical_specialty, HbA1c, Endocrine Disorders, 030209 endocrinology & metabolism, Carbohydrate metabolism, Blood Plasma, 03 medical and health sciences, Diagnostic Medicine, Internal medicine, Diabetes mellitus, Genetics, medicine, Diabetes Mellitus, Hemoglobin, Non-coding RNA, Biology and life sciences, business.industry, lcsh:R, Proteins, nutritional and metabolic diseases, medicine.disease, Gene regulation, Circulating MicroRNA, Metabolism, 030104 developmental biology, chemistry, Metabolic Disorders, Glucose Tolerance Tests, RNA, lcsh:Q, Gene expression, business
Abstract

A major unmet medical need to better manage Type 2 Diabetes (T2D) is the accurate disease prediction in subjects who show glucose dysmetabolism, but are not yet diagnosed as diabetic. We investigated the possibility to predict/monitor the progression to T2D in these subjects by retrospectively quantifying blood circulating microRNAs in plasma of subjects with i) normal glucose tolerance (NGT, n = 9); ii) impaired glucose tolerance (IGT, n = 9), divided into non-progressors (NP, n = 5) and progressors (P, n = 4) based on subsequent diabetes occurrence, and iii) newly diagnosed T2D (n = 9). We found that impaired glucose tolerance associated with a global increase of plasma circulating microRNAs. While miR-148 and miR-222 were specifically modulated in diabetic subjects and correlated with parameters of glucose tolerance, the most accentuated microRNA dysregulation was found in NP IGT subjects, with increased level of miR-122, miR-99 and decreased level of let-7d, miR-18a, miR-18b, miR-23a, miR-27a, miR-28 and miR-30d in comparison with either NGT or T2D. Interestingly, several of these microRNAs significantly correlated with parameters of cholesterol metabolism. In conclusion, we observed the major perturbation of plasma circulating microRNA in NP pre-diabetic subjects and identified a unique microRNA profile that may become helpful in predicting diabetic development.

Published
2017

36. Microfluidic Synthesis of Hybrid TiO 2 -Anisotropic Gold Nanoparticles with Visible and Near-Infrared Activity.

Author

Marelli M, Bossola F, Spinetti G, Sangalli E, Santo VD, Psaro R, and Polito L

Abstract

Anisotropic gold nanoparticles (AuNPs), with their unique physical and optical properties, are emerging as smart and key nanomaterials and are being exploited in many crucial fields. To further improve their range of action, anisotropic AuNPs have been coupled with semiconductors, mainly TiO 2 (titania), receiving great interest as powerful platforms both in biomedicine and in catalytic applications. Such hybrid nanoparticles show new properties that arise from the synergic action of the components and rely on NP size, morphology, and arrangement. Therefore, continuous advances in design and fabrication of new hybrid titania@gold NPs (TiO 2 @AuNPs) are urgent and highly desirable. Here, we propose an effective protocol to produce multibranched AuNPs covered by a controlled TiO 2 thin layer, exploiting a one-pot microfluidic process. The proposed method allows the in-flow and reliable synthesis of titania-functionalized-anisotropic gold nanoparticles by avoiding the use of toxic surfactants and controlling the titania shell formation. TiO 2 @AuNPs have been fully characterized in terms of morphology, stability, and biocompatibility, and their activity in photocatalysis has been tested and verified.

Published
2020
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