@article{%a1.%Y_161,

title = {CAR+ extracellular vesicles predict ICANS in patients with B cell lymphomas treated with CD19-directed CAR T cells},

author = {Storci G and De Felice F and Ricci F and Santi S and Messelodi D and Bertuccio SN and Laprovitera N and Dicataldo M and Rossini L and De Matteis S and Casadei B and Vaglio F and Ursi M and Barbato F and Roberto M and Guarino M and Asioli GM and Arpinati M and Cortelli P and Maffini E and Tomassini E and Tassoni M and Cavallo C and Iannotta F and Naddeo M and Tazzari PL and Dan E and Pellegrini C and Guadagnuolo S and Carella M and Sinigaglia B and Pirazzini C and Severi C and Garagnani P and Kwiatkowska KM and Ferracin M and Zinzani PL and Bonafe’ M and Bonifazi F.},

url = {https://www.jci.org/articles/view/173096},

doi = {10.1172/JCI173096},

year  = {2024},

date = {2024-08-06},

journal = {Journal of clinical investigation},

volume = {134},

issue = {14},

pages = {e173096},

abstract = {Background: Predicting Immune-effector Cell Associated Neurotoxicity Syndrome (ICANS) in patients infused with Chimeric Antigen Receptor T cells (CAR-T) is still a conundrum. This complication, thought to be consequent to CAR-T cell activation, arises a few days after infusion, when circulating CAR-T cells are scarce and specific CAR-T cell-derived biomarkers are lacking. Methods: Human CD19.CAR-T cells were generated to gain insight into CAR+ extracellular vesicle (CAR+EV) release upon target engagement. A prospective cohort of 100 B-cell lymphoma patients infused with approved CD19.CAR-T cell products (axi-cel, brexu-cel and tisa-cel) was assessed for plasma CAR+EVs as potential biomarkers of in vivo CD19.CAR-T cell activation and predictors of ICANS. Human induced pluripotent stem cells (iPSCs)-derived neural cells were used as a model for CAR+EV-induced neurotoxicity. Results: In vitro, exosome-like CAR+EVs were released by CD19.CAR-T cells upon target engagement. In vivo, CAR+EVs were detectable as early as 1 hour in the plasma of patients. A concentration > 132.8 CAR+EVs/μl at hour +1 or > 224.5 CAR+EVs/μl at day +1 predicted ICANS in advance of 4 days, with a sensitivity up to 96.55% and a specificity up to 80.36%, outperforming other potential ICANS predictors. Enolase 2 (ENO2+) nanoparticles were released by iPSCs-derived neural cells upon CAR+EVs exposure and were increased in the plasma of ICANS patients. Conclusions: These results convey that plasma CAR+EVs are an immediate signal of CD19.CAR-T cell activation, are suitable predictors of neurotoxicity, and may be involved in ICANS pathogenesis.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{%a1.%Y_151,

title = {Collagen VI Deficiency Impairs Tendon Fibroblasts Mechanoresponse in Ullrich Congenital Muscular Dystrophy},

author = {Cenni V and Sabatelli P and Di Martino A and Merlini L and Antoniel M and Squarzoni S and Neri S and Santi S and Metti S and Bonaldo P and Faldini C.},

url = {https://www.mdpi.com/2073-4409/13/5/378},

doi = {10.3390/cells13050378},

year  = {2024},

date = {2024-08-06},

journal = {Cells},

volume = {13},

issue = {5},

pages = {378},

abstract = {The pericellular matrix (PCM) is a specialized extracellular matrix that surrounds cells. Interactions with the PCM enable the cells to sense and respond to mechanical signals, triggering a proper adaptive response. Collagen VI is a component of muscle and tendon PCM. Mutations in collagen VI genes cause a distinctive group of inherited skeletal muscle diseases, and Ullrich congenital muscular dystrophy (UCMD) is the most severe form. In addition to muscle weakness, UCMD patients show structural and functional changes of the tendon PCM. In this study, we investigated whether PCM alterations due to collagen VI mutations affect the response of tendon fibroblasts to mechanical stimulation. By taking advantage of human tendon cultures obtained from unaffected donors and from UCMD patients, we analyzed the morphological and functional properties of cellular mechanosensors. We found that the length of the primary cilia of UCMD cells was longer than that of controls. Unlike controls, in UCMD cells, both cilia prevalence and length were not recovered after mechanical stimulation. Accordingly, under the same experimental conditions, the activation of the Hedgehog signaling pathway, which is related to cilia activity, was impaired in UCMD cells. Finally, UCMD tendon cells exposed to mechanical stimuli showed altered focal adhesions, as well as impaired activation of Akt, ERK1/2, p38MAPK, and mechanoresponsive genes downstream of YAP. By exploring the response to mechanical stimulation, for the first time, our findings uncover novel unreported mechanistic aspects of the physiopathology of UCMD-derived tendon fibroblasts and point at a role for collagen VI in the modulation of mechanotransduction in tendons.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{%a1.%Y,

title = {Desmin and Plectin Recruitment to the Nucleus and Nuclei Orientation Are Lost in Emery-Dreifuss Muscular Dystrophy Myoblasts Subjected to Mechanical Stimulation},

author = {Cenni V and Evangelisti C and Santi S and Sabatelli P and Neri S and Cavallo M and Lattanzi G and Mattioli E},

url = {https://www.mdpi.com/2073-4409/13/2/162},

doi = {10.3390/cells13020162},

year  = {2024},

date = {2024-01-31},

journal = {Cells},

volume = {13},

issue = {2},

pages = {162},

abstract = {n muscle cells subjected to mechanical stimulation, LINC complex and cytoskeletal proteins are basic to preserve cellular architecture and maintain nuclei orientation and positioning. In this context, the role of lamin A/C remains mostly elusive. This study demonstrates that in human myoblasts subjected to mechanical stretching, lamin A/C recruits desmin and plectin to the nuclear periphery, allowing a proper spatial orientation of the nuclei. Interestingly, in Emery-Dreifuss Muscular Dystrophy (EDMD2) myoblasts exposed to mechanical stretching, the recruitment of desmin and plectin to the nucleus and nuclear orientation were impaired, suggesting that a functional lamin A/C is crucial for the response to mechanical strain. While describing a new mechanism of action headed by lamin A/C, these findings show a structural alteration that could be involved in the onset of the muscle defects observed in muscular laminopathies.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{%a1.%Y_139,

title = {Engagement of CD99 activates distinct programs in Ewing sarcoma and macrophages},

author = {Manara MC and Manferdini C and Cristalli C and Carrabotta M and Santi S and De Feo A and Caldoni G and Pasello M and Landuzzi L and Lollini PL and Salamanna F and Dominici S and Fiori V and Magnani M and Lisignoli G and Scotlandi K},

url = {https://aacrjournals.org/cancerimmunolres/article/doi/10.1158/2326-6066.CIR-23-0440/731506/Engagement-of-CD99-activates-distinct-programs-in},

doi = {10.1158/2326-6066.CIR-23-0440},

year  = {2024},

date = {2024-02-12},

journal = {Cancer immunology research},

volume = {12},

issue = {2},

pages = {247-260},

abstract = {Ewing sarcoma (EWS) is the second most common pediatric bone tumor. The EWS tumor microenvironment is largely recognized as immune-cold, with macrophages being the most abundant immune cells and their presence associated with worse patient prognosis. Expression of CD99 is a hallmark of EWS cells, and its targeting induces inhibition of EWS tumor growth through a poorly understood mechanism. In this study, we analyzed CD99 expression and functions on macrophages and investigated whether the concomitant targeting of CD99 on both tumor and macrophages could explain the inhibitory effect of this approach against EWS. Targeting CD99 on EWS cells downregulated expression of the "don't eat-me" CD47 molecule but increased levels of the "eat-me" phosphatidyl serine and calreticulin molecules on the outer leaflet of the tumor cell membrane, triggering phagocytosis and digestion of EWS cells by macrophages. In addition, CD99 ligation induced reprogramming of undifferentiated M0 macrophages and M2-like macrophages toward the inflammatory M1-like phenotype. These events resulted in the inhibition of EWS tumor growth. Thus, this study reveals what we believe to be a previously unrecognized function of CD99, which engenders a virtuous circle that delivers intrinsic cell death signals to EWS cells, favors tumor cell phagocytosis by macrophages, and promotes the expression of various molecules and cytokines, which are pro-inflammatory and usually associated with tumor regression. This raises the possibility that CD99 may be involved in boosting the antitumor activity of macrophages.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{%a1.%Y_153,

title = {Replicative senescence and high glucose induce the accrual of self-derived cytosolic nucleic acids in human endothelial cells},

author = {Ramini D and Giuliani A and Kwiatkowska KM and Guescini M and Storci G and Mensa' E and Recchioni R and Xumerle L and Zago E and Sabbatinelli J and Santi S and Garagnani P and Bonafe' M and Olivieri F.},

url = {https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11032409/},

doi = {10.1038/s41420-024-01954-z},

year  = {2024},

date = {2024-05-28},

urldate = {2024-05-28},

journal = {Cell death discovery},

volume = {10},

number = {184},

issue = {1},

abstract = {Recent literature shows that loss of replicative ability and acquisition of a proinflammatory secretory phenotype in senescent cells is coupled with the build-in of nucleic acids in the cytoplasm. Its implication in human age-related diseases is under scrutiny. In human endothelial cells (ECs), we assessed the accumulation of intracellular nucleic acids during in vitro replicative senescence and after exposure to high glucose concentrations, which mimic an in vivo condition of hyperglycemia. We showed that exposure to high glucose induces senescent-like features in ECs, including telomere shortening and proinflammatory cytokine release, coupled with the accrual in the cytoplasm of telomeres, double-stranded DNA and RNA (dsDNA, dsRNA), as well as RNA:DNA hybrid molecules. Senescent ECs showed an activation of the dsRNA sensors RIG-I and MDA5 and of the DNA sensor TLR9, which was not paralleled by the involvement of the canonical (cGAS) and non-canonical (IFI16) activation of the STING pathway. Under high glucose conditions, only a sustained activation of TLR9 was observed. Notably, senescent cells exhibit increased proinflammatory cytokine (IL-1β, IL-6, IL-8) production without a detectable secretion of type I interferon (IFN), a phenomenon that can be explained, at least in part, by the accumulation of methyl-adenosine containing RNAs. At variance, exposure to exogenous nucleic acids enhances both IL-6 and IFN-β1 expression in senescent cells. This study highlights the accrual of cytoplasmic nucleic acids as a marker of senescence-related endothelial dysfunction, that may play a role in dysmetabolic age-related diseases.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{%a1.%Y_133,

title = {Aberrant MET activation impairs perinuclear actin cap organization with YAP1 cytosolic relocation},

author = {Sgarzi M and Mazzeschi M and Santi S and Montacci E and Panciera T and Ferlizza E and Girone C and Morselli A and Gelfo V and Kuhre RS and Cavallo C and Valente S and Pasquinelli G and Gyorffy B and D'Uva G and Romaniello D and Lauriola M},

url = {https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10576810/},

doi = {10.1038/s42003-023-05411-y},

year  = {2023},

date = {2023-08-23},

urldate = {2023-08-23},

journal = {Communications biology},

volume = {6},

issue = {1},

pages = {1044},

abstract = {Little is known about the signaling network responsible for the organization of the perinuclear actin cap, a recently identified structure holding unique roles in the regulation of nuclear shape and cell directionality. In cancer cells expressing a constitutively active MET, we show a rearrangement of the actin cap filaments, which crash into perinuclear patches associated with spherical nuclei, meandering cell motility and inactivation of the mechano-transducer YAP1. MET ablation is sufficient to reactivate YAP1 and restore the cap, leading to enhanced directionality and flattened nuclei. Consistently, the introduction of a hyperactive MET in normal epithelial cells, enhances nuclear height and alters the cap organization, as also confirmed by TEM analysis. Finally, the constitutively active YAP1 mutant YAP5SA is able to overcome the effects of oncogenic MET. Overall, our work describes a signaling axis empowering MET-mediated YAP1 dampening and actin cap misalignment, with implications for nuclear shape and cell motility.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{%a1.%Yb_78,

title = {Impaired synaptic plasticity in an animal model of autism exhibiting early hippocampal GABAergic-BDNF/TrkB signaling alterations},

author = {Sgritta M and Vignoli B and Pimpinella D and Griguoli M and Santi S and Bialowas A and Wiera G and Zacchi P and Malerba F and Marchetti C and Canossa M and Cherubini E},

url = {https://www.sciencedirect.com/science/article/pii/S2589004222020016?via%3Dihub},

doi = {10.1016/j.isci.2022.105728},

year  = {2023},

date = {2023-03-09},

journal = {iScience},

volume = {26},

issue = {1},

pages = {105728},

abstract = {In Neurodevelopmental Disorders, alterations of synaptic plasticity may trigger structural changes in neuronal circuits involved in cognitive functions. This hypothesis was tested in mice carrying the human R451C mutation of Nlgn3 gene (NLG3R451C KI), found in some families with autistic children. To this aim, the spike time dependent plasticity (STDP) protocol was applied to immature GABAergic Mossy Fibers (MF)-CA3 connections in hippocampal slices from NLG3R451C KI mice. These animals failed to exhibit STD-LTP, an effect that persisted in adulthood when these synapses became glutamatergic. Similar results were obtained in mice lacking the Nlgn3 gene (NLG3 KO mice), suggesting a loss of function. The loss of STD-LTP was associated with a premature shift of GABA from the depolarizing to the hyperpolarizing direction, a reduced BDNF availability and TrkB phosphorylation at potentiated synapses. These effects may constitute a general mechanism underlying cognitive deficits in those forms of Autism caused by synaptic dysfunctions.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{%a1.%Yb_79,

title = {Pre-transplant CD69+ extracellular vesicles are negatively correlated with active ATLG serum levels and associate with the onset of GVHD in allogeneic HSCT patients},

author = {Storci G and Barbato F and Ricci F and Tazzari PL and De Matteis S and Tomassini E and Dicataldo M and Laprovitera N and Arpinati M and Ursi M and Maffini E and Campanini E and Dan E and Manfroi S and Santi S and Ferracin M and Bonafe M and Bonifazi F},

url = {https://www.frontiersin.org/articles/10.3389/fimmu.2022.1058739/full},

doi = {10.3389/fimmu.2022.1058739},

year  = {2023},

date = {2023-03-09},

urldate = {2023-03-09},

journal = {Frontiers in immunology},

volume = {13},

abstract = {Graft versus host disease (GVHD) is a major complication of allogeneic hematopoietic stem cell transplantation (HSCT). Rabbit anti-T lymphocyte globulin (ATLG) in addition to calcineurin inhibitors and antimetabolites is a suitable strategy to prevent GVHD in several transplant settings. Randomized studies already demonstrated its efficacy in terms of GVHD prevention, although the effect on relapse remains the major concern for a wider use. Tailoring of ATLG dose on host characteristics is expected to minimize its side effects (immunological reconstitution, relapse, and infections). Here, day -6 to day +15 pharmacokinetics of active ATLG serum level was first assayed in an explorative cohort of 23 patients by testing the ability of the polyclonal serum to bind antigens on human leukocytes. Significantly lower levels of serum active ATLG were found in the patients who developed GVHD (ATLG_AUCCD45: 241.52 ± 152.16 vs. 766.63 +/- 283.52 (μg*day)/ml, p = 1.46e-5). Consistent results were obtained when the ATLG binding capacity was assessed on CD3+ and CD3+/CD4+ T lymphocytes (ATLG_AUCCD3: 335.83 ± 208.15 vs. 903.54 ± 378.78 (μg*day)/ml, p = 1.92e-4; ATLG_AUCCD4: 317.75 ± 170.70 vs. 910.54 ± 353.35 (μg*day)/ml, p = 3.78e-5. Concomitantly, at pre-infusion time points, increased concentrations of CD69+ extracellular vesicles (EVs) were found in patients who developed GVHD (mean fold 9.01 ± 1.33; p = 2.12e-5). Consistent results were obtained in a validation cohort of 12 additional ATLG-treated HSCT patients. Serum CD69+ EVs were mainly represented in the nano (i.e. 100 nm in diameter) EV compartment and expressed the leukocyte marker CD45, the EV markers CD9 and CD63, and CD103, a marker of tissue-resident memory T cells. The latter are expected to set up a host pro-inflammatory cell compartment that can survive in the recipient for years after conditioning regimen and contribute to GVHD pathogenesis. In summary, high levels of CD69+ EVs are significantly correlated with an increased risk of GVHD, and they may be proposed as a tool to tailor ATLG dose for personalized GVHD prevention.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{%a1.%Yb_112,

title = {Targeting the Interplay of Independent Cellular Pathways and Immunity: A Challenge in Cancer Immunotherapy},

author = {Lauriola A and Davalli P and Marverti G and Santi S and Caporali A and D'Arca D},

url = {https://www.mdpi.com/2072-6694/15/11/3009},

doi = {10.3390/cancers15113009},

year  = {2023},

date = {2023-08-08},

journal = {Cancers-Basel},

volume = {15},

issue = {11},

pages = {3009},

abstract = {Immunotherapy is a cancer treatment that exploits the capacity of the body's immune system to prevent, control, and remove cancer. Immunotherapy has revolutionized cancer treatment and significantly improved patient outcomes for several tumor types. However, most patients have not benefited from such therapies yet. Within the field of cancer immunotherapy, an expansion of the combination strategy that targets independent cellular pathways that can work synergistically is predicted. Here, we review some consequences of tumor cell death and increased immune system engagement in the modulation of oxidative stress and ubiquitin ligase pathways. We also indicate combinations of cancer immunotherapies and immunomodulatory targets. Additionally, we discuss imaging techniques, which are crucial for monitoring tumor responses during treatment and the immunotherapy side effects. Finally, the major outstanding questions are also presented, and directions for future research are described.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{%a1.%Ybj,

title = {Exosomes Recovered From the Plasma of COVID-19 Patients Expose SARS-CoV-2 Spike-Derived Fragments and Contribute to the Adaptive Immune Response},

author = {Pesce E and Manfrini N and Cordiglieri C and Santi S and Bandera A and Gobbini A and Gruarin P and Favalli A and Bombaci M and Cuomo A and Collino F and Cricri' G and Ungaro R and Lombardi A and Mangioni D and Muscatello A and Aliberti S and Blasi F and Gori A and Abrignani S and De Francesco R and Biffo S and Grifantini R},

url = {http://www.scopus.com/inward/record.url?eid=2-s2.0-85123854300&partnerID=q2rCbXpz},

doi = {10.3389/fimmu.2021.785941},

year  = {2022},

date = {2022-03-21},

journal = {Frontiers in immunology},

volume = {12},

pages = {7859},

abstract = {Coronavirus disease 2019 (COVID-19) is an infectious disease caused by beta-coronavirus severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) that has rapidly spread across the globe starting from February 2020. It is well established that during viral infection, extracellular vesicles become delivery/presenting vectors of viral material. However, studies regarding extracellular vesicle function in COVID-19 pathology are still scanty. Here, we performed a comparative study on exosomes recovered from the plasma of either MILD or SEVERE COVID-19 patients. We show that although both types of vesicles efficiently display SARS-CoV-2 spike-derived peptides and carry immunomodulatory molecules, only those of MILD patients are capable of efficiently regulating antigen-specific CD4+ T-cell responses. Accordingly, by mass spectrometry, we show that the proteome of exosomes of MILD patients correlates with a proper functioning of the immune system, while that of SEVERE patients is associated with increased and chronic inflammation. Overall, we show that exosomes recovered from the plasma of COVID-19 patients possess SARS-CoV-2-derived protein material, have an active role in enhancing the immune response, and possess a cargo that reflects the pathological state of patients in the acute phase of the disease.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{%a1.%Ybv,

title = {Small Extracellular Vesicles from Inflamed Adipose Derived Stromal Cells Enhance the NF-κB-Dependent Inflammatory/Catabolic Environment of Osteoarthritis. },

author = {Cavallo C and Merli G and Zini N and {D'Adamo} S and Cattini L and Guescini M and Grigolo B and Di Martino A and Santi S and Borzì RM and Filardo G},

url = {https://www.hindawi.com/journals/sci/2022/9376338/},

doi = {10.1155/2022/9376338},

year  = {2022},

date = {2022-08-11},

urldate = {2022-08-11},

journal = {Stem cells international},

volume = {2022},

pages = {9376338},

abstract = {The last decade has seen exponentially growing efforts to exploit the effects of adipose derived stromal cells (ADSC) in the treatment of a wide range of chronic degenerative diseases, including osteoarthritis (OA), the most prevalent joint disorder. In the perspective of developing a cell-free advanced therapy medicinal product, a focus has been recently addressed to the ADSC secretome that lends itself to an allogeneic use and can be further dissected for the selective purification of small extracellular vesicles (sEVs). sEVs can act as "biological drug carriers" to transfer information that mirror the pathophysiology of the providing cells. This is important in the clinical perspective where many OA patients are also affected by the metabolic syndrome (MetS). ADSC from MetS OA patients are dysfunctional and "inflammatory" primed within the adipose tissue. To mimic this condition, we exposed ADSC to IL-1β, and then we investigated the effects of the isolated sEVs on chondrocytes and synoviocytes, either cultured separately or in co-culture, to tease out the effects of these "IL-1β primed sEVs" on gene and protein expression of major inflammatory and catabolic OA markers. In comparison with sEVs isolated from unstimulated ADSC, the IL-1β primed sEVs were able to propagate NF-κB activation in bystander joint cells. The effects were more prominent on synoviocytes, possibly because of a higher expression of binding molecules such as CD44. These findings call upon a careful characterization of the "inflammatory fingerprint" of ADSC to avoid the transfer of an unwanted message as well as the development of in vitro "preconditioning" strategies able to rescue the antiinflammatory/anticatabolic potential of ADSC-derived sEVs.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{%a1.%Yb_30,

title = {The autocrine loop of ALK receptor and ALKAL2 ligand is an actionable target in consensus molecular subtype 1 colon cancer},

author = {Mazzeschi M and Sgarzi M and Romaniello D and Gelfo V and Cavallo C and Ambrosi F and Morselli A and Miano C and Laprovitera N and Girone C and Ferracin M and Santi S and Rihawi K and Ardizzoni A and Fiorentino M and D'Uva G and Győrffy B and Palmer R and Lauriola M},

url = {https://jeccr.biomedcentral.com/articles/10.1186/s13046-022-02309-1},

doi = {10.1186/s13046-022-02309-1},

year  = {2022},

date = {2022-08-19},

journal = {Journal of experimental & clinical cancer research},

volume = {41},

issue = {1},

pages = {113},

abstract = {Background: In the last years, several efforts have been made to classify colorectal cancer (CRC) into well-defined molecular subgroups, representing the intrinsic inter-patient heterogeneity, known as Consensus Molecular Subtypes (CMSs). Methods: In this work, we performed a meta-analysis of CRC patients stratified into four CMSs. We identified a negative correlation between a high level of anaplastic lymphoma kinase (ALK) expression and relapse-free survival, exclusively in CMS1 subtype. Stemming from this observation, we tested cell lines, patient-derived organoids and mice with potent ALK inhibitors, already approved for clinical use. Results: ALK interception strongly inhibits cell proliferation already at nanomolar doses, specifically in CMS1 cell lines, while no effect was found in CMS2/3/4 groups. Furthermore, in vivo imaging identified a role for ALK in the dynamic formation of 3D tumor spheroids. Consistently, ALK appeares constitutively phosphorylated in CMS1, and it signals mainly through the AKT axis. Mechanistically, we found that CMS1 cells display several copies of ALKAL2 ligand and ALK-mRNAs, suggesting an autocrine loop mediated by ALKAL2 in the activation of ALK pathway, responsible for the invasive phenotype. Consequently, disruption of ALK axis mediates the pro-apoptotic action of CMS1 cell lines, both in 2D and 3D and enhanced cell-cell adhesion and e-cadherin organization. In agreement with all these findings, the ALK signature encompassing 65 genes statistically associated with worse relapse-free survival in CMS1 subtype. Finally, as a proof of concept, the efficacy of ALK inhibition was demonstrated in both patient-derived organoids and in tumor xenografts in vivo. Conclusions: Collectively, these findings suggest that ALK targeting may represent an attractive therapy for CRC, and CMS classification may provide a useful tool to identify patients who could benefit from this treatment. These findings offer rationale and pharmacological strategies for the treatment of CMS1 CRC.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{%a1:%Yb,

title = {A new method for the study of biophysical and morphological parameters in 3D cell cultures: Evaluation in LoVo spheroids treated with crizotinib},

author = {Sargenti A and Musmeci F and Cavallo C and Mazzeschi M and Bonetti S and Pasqua S and Bacchi F and Filardo G and Gazzola D and Lauriola M and Santi S},

url = {https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0252907},

doi = {10.1371/journal.pone.0252907},

year  = {2021},

date = {2021-06-10},

journal = {PLoS One},

volume = {16},

number = {6},

pages = {PLoS One},

abstract = {Three-dimensional (3D) culture systems like tumor spheroids represent useful in vitro models for drug screening and more broadly for cancer biology research, but the generation of uniform populations of spheroids remains challenging. The possibility to properly characterize spheroid properties would increase the reliability of these models. To address this issue different analysis were combined: i) a new device and relative analytical method for the accurate, simultaneous, and rapid measurement of mass density, weight, and size of spheroids, ii) confocal imaging, and iii) protein quantification, in a clinically relevant 3D model. The LoVo colon cancer cell line forming spheroids, treated with crizotinib (CZB) an ATP-competitive small-molecule inhibitor of the receptor tyrosine kinases, was employed to study and assess the correlation between biophysical and morphological parameters in both live and fixed cells. The new fluidic-based measurements allowed a robust phenotypical characterization of the spheroids structure, offering insights on the spheroids bulk and an accurate measurement of the tumor density. This analysis helps overcome the technical limits of the imaging that hardly penetrates the thickness of 3D structures. Accordingly, we were able to document that CZB treatment has an impact on mass density, which represents a key marker characterizing cancer cell treatment. Spheroid culture is the ultimate technology in drug discovery and the adoption of such precise measurement of the tumor characteristics can represent a key step forward for the accurate testing of treatment's potential in 3D in vitro models.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{%a1:%Ybv_28,

title = {Astrocytic microdomains from mouse cortex gain molecular control over long-term information storage and memory retention},

author = {Vignoli B and Sansevero G and Sasi M and Rimondini R and Blum R and Bonaldo V and Biasini E and Santi S and Berardi N and Lu B and Canossa M},

url = {https://www.nature.com/articles/s42003-021-02678-x},

doi = {10.1038/s42003-021-02678-x},

year  = {2021},

date = {2021-10-28},

journal = {Communications biology},

volume = {4},

number = {1},

pages = {1152},

abstract = {Memory consolidation requires astrocytic microdomains for protein recycling; but whether this lays a mechanistic foundation for long-term information storage remains enigmatic. Here we demonstrate that persistent synaptic strengthening invited astrocytic microdomains to convert initially internalized (pro)-brain-derived neurotrophic factor (proBDNF) into active prodomain (BDNFpro) and mature BDNF (mBDNF) for synaptic re-use. While mBDNF activates TrkB, we uncovered a previously unsuspected function for the cleaved BDNFpro, which increases TrkB/SorCS2 receptor complex at post-synaptic sites. Astrocytic BDNFpro release reinforced TrkB phosphorylation to sustain long-term synaptic potentiation and to retain memory in the novel object recognition behavioral test. Thus, the switch from one inactive state to a multi-functional one of the proBDNF provides post-synaptic changes that survive the initial activation. This molecular asset confines local information storage in astrocytic microdomains to selectively support memory circuits.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{%a1:%Ybvwd,

title = {Basal and IL-1beta enhanced chondrocyte chemotactic activity on monocytes are co-dependent on both IKKalpha and IKKbeta NF-kappaB activating kinases},

author = {Olivotto E and Minguzzi M and D'Adamo S and Astolfi A and Santi S and Uguccioni M and Marcu KB and Borzi' RM},

url = {https://www.nature.com/articles/s41598-021-01063-2},

doi = {10.1038/s41598-021-01063-2},

year  = {2021},

date = {2021-11-08},

urldate = {2021-11-08},

journal = {Scientific reports},

volume = {11},

number = {1},

pages = {21697},

abstract = {IKKalpha and IKKbeta are essential kinases for activating NF-kappaB transcription factors that regulate cellular differentiation and inflammation. By virtue of their small size, chemokines support the crosstalk between cartilage and other joint compartments and contribute to immune cell chemotaxis in osteoarthritis (OA). Here we employed shRNA retroviruses to stably and efficiently ablate the expression of each IKK in primary OA chondrocytes to determine their individual contributions for monocyte chemotaxis in response to chondrocyte conditioned media. Both IKKalpha and IKKbeta KDs blunted both the monocyte chemotactic potential and the protein levels of CCL2/MCP-1, the chemokine with the highest concentration and the strongest association with monocyte chemotaxis. These findings were mirrored by gene expression analysis indicating that the lowest levels of CCL2/MCP-1 and other monocyte-active chemokines were in IKKαKD cells under both basal and IL-1beta stimulated conditions. We find that in their response to IL-1beta stimulation IKKalphaKD primary OA chondrocytes have reduced levels of phosphorylated NFkappaB p65pSer536 and H3pSer10. Confocal microscopy analysis revealed co-localized p65 and H3pSer10 nuclear signals in agreement with our findings that IKKalphaKD effectively blunts their basal level and IL-1beta dependent increases. Our results suggest that IKKalpha could be a novel OA disease target.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{%a1:%Ybu,

title = {Exploring the anticancer effects of standardized extracts of poplar-type propolis: In vitro cytotoxicity toward cancer and normal cell lines},

author = {Campoccia D and Ravaioli S and Santi S and Mariani V and Santarcangelo C and De Filippis A and Montanaro L and Arciola CR and Daglia M},

url = {https://www.sciencedirect.com/science/article/pii/S0753332221006776?via%3Dihub},

doi = {10.1016/j.biopha.2021.111895},

year  = {2021},

date = {2021-08-25},

urldate = {2021-08-25},

journal = {Biomedicine & pharmacotherapy},

volume = {141},

pages = {111895},

abstract = {Propolis was shown to exert antimicrobial, antioxidant, anti-inflammatory, and anticancer activities. Its composition is influenced by seasonal, climatic and phytogeographic conditions. Further variability derives from the extraction methods. Multi Dynamic Extraction Method (MED) has been recently proposed to improve extracts reproducibility. Here, the cytotoxic/anticancer activity of three MED extracts of poplar-type propolis was assayed on human promyelocytic leukaemia HL60, human monocytic leukaemia THP-1, human osteosarcoma MG63, murine fibroblast L929 and human mesenchymal cells (hMSCs). As far as we are aware of, MG63 cells have never been challenged with propolis before, while few studies have so far addressed the effects of propolis on non-tumor cell lines. Consistent results were observed for all propolis preparations. The extracts turned out mildly cytotoxic toward cancer cells, in particular osteosarcoma cells (IC50: 81.9-86.7 µg/ml). Nonetheless, cytotoxicity was observed also in non-tumor L929 cells, with an even lower IC50. hMSCs demonstrated the lowest sensitivity to propolis (IC50: 258.3-287.2 µg/ml). In THP-1 cells, extracts were found to stimulate apoptosis caspase 3/7 activity. The IC50 values observed with osteosarcoma and leukaemia cells do not support a relevant cytotoxicity (as the figures abundantly exceeded 30 µg/ml), despites some selective activity exhibited with HL60 cells. The results confirm the validity of the extraction method, emphasizing the need to assess the selectivity of the interaction with cancer cells when screening for anticancer-drug candidates.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{%a1:%Y__494,

title = {Morphological study of TNPO3 and SRSF1 interaction during myogenesis by combining confocal, structured illumination and electron microscopy analysis. },

author = {Costa R and Rodia MT and Zini N and Pegoraro V and Marozzo R and Capanni C and Angelini C and Lattanzi G and Santi S and Cenacchi G},

url = {https://link.springer.com/article/10.1007/s11010-020-04023-y},

doi = {10.1007/s11010-020-04023-y},

year  = {2021},

date = {2021-03-09},

urldate = {2021-03-09},

journal = {Molecular and cellular biochemistry},

volume = {476},

number = {4},

pages = {1797-1811},

abstract = {Transportin3 (TNPO3) shuttles the SR proteins from the cytoplasm to the nucleus. The SR family includes essential splicing factors, such as SRSF1, that influence alternative splicing, controlling protein diversity in muscle and satellite cell differentiation. Given the importance of alternative splicing in the myogenic process and in the maintenance of healthy muscle, alterations in the splicing mechanism might contribute to the development of muscle disorders. Combining confocal, structured illumination and electron microscopy, we investigated the expression of TNPO3 and SRSF1 during myogenesis, looking at nuclear and cytoplasmic compartments. We investigated TNPO3 and its interaction with SRSF1 and we observed that SRSF1 remained mainly localized in the nucleus, while TNPO3 decreased in the cytoplasm and was strongly clustered in the nuclei of differentiated myotubes. In conclusion, combining different imaging techniques led us to describe the behavior of TNPO3 and SRSF1 during myogenesis, showing that their dynamics follow the myogenic process and could influence the proteomic network necessary during myogenesis. The combination of different high-, super- and ultra-resolution imaging techniques led us to describe the behavior of TNPO3 and its interaction with SRSF1, looking at nuclear and cytoplasmic compartments. These observations represent a first step in understanding the role of TNPO3 and SRFSF1 in complex mechanisms, such as myogenesis.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{%a1:%Y__497,

title = {Oxidative stress-induced DNA damage and repair in primary human osteoarthritis chondrocytes: focus on IKKα and the DNA Mismatch Repair System},

author = {Neri S and Guidotti S and Bini C and Pelotti S and D'Adamo S and Minguzzi M and Platano D and Santi S and Mariani E and Cattini L and Borzì RM},

url = {https://www.sciencedirect.com/science/article/pii/S0891584921001118?via%3Dihub},

doi = {10.1016/j.freeradbiomed.2021.02.020},

year  = {2021},

date = {2021-03-09},

journal = {Free radical biology & medicine},

volume = {166},

number = {212-225},

abstract = {During osteoarthritis development, chondrocytes are subjected to a functional derangement. This increases their susceptibility to stressful conditions such as oxidative stress, a characteristic of the aging tissue, which can further provoke extrinsic senescence by DNA damage responses. It was previously observed that IκB kinase α knockdown increases the replicative potential of primary human OA chondrocytes cultured in monolayer and the survival of the same cells undergoing hypertrophic-like differentiation in 3-D. In this paper we investigated whether IKKα knockdown could modulate oxidative stress-induced senescence of OA chondrocytes undergoing a DDR and particularly the involvement in this process of the DNA Mismatch repair system, the principal mechanism for repair of replicative and recombinational errors, devoted to genomic stability maintenance in actively replicating cells. This repair system is also implicated in oxidative stress-mediated DNA damage repair. We analyzed microsatellite instability and expression of the mismatch repair components in human osteoarthritis chondrocytes after IKKα knockdown and H2O2 exposure. Only low MSI levels and incidence were detected and exclusively in IKKα proficient cells. Moreover, we found that IKKα proficient and deficient chondrocytes differently regulated MMR proteins after oxidative stress, both at mRNA and protein level, suggesting a reduced susceptibility of IKKα deficient cells. Our data suggest an involvement of the MMR system in the response to oxidative stress that tends to be more efficient in IKKαKD cells. This argues for a partial contribution of the MMR system to the better ability to recover DNA damage already observed in these cells.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{%a1:%Y_433,

title = {Protein aggregates and autophagy involvement in a family with a mutation in Z-band alternatively spliced PDZ-motif protein.},

author = {Cassandrini D and Merlini L and Pilla F and Cenni V and Santi S and Faldini C and Santorelli FM and Sabatelli P},

url = {https://www.nmd-journal.com/article/S0960-8966(20)30673-8/fulltext},

doi = {10.1016/j.nmd.2020.11.008},

year  = {2021},

date = {2021-03-09},

journal = {Neuromuscular disorders},

volume = {S0960-8966},

number = {20},

pages = {30673-30678},

abstract = {Z-band alternatively spliced PDZ-motif protein (ZASP) is a sarcomeric component expressed both in cardiac and skeletal muscles. Mutations in the LDB3/ZASP gene cause cardiomyopathy and myofibrillar myopathy. We describe a c.76C>T / p.[Pro26Ser] mutation in the PDZ motif of LDB3/ZASP in two siblings exhibiting late-onset myopathy with axial, proximal and distal muscles involvement and marked variability in clinical severity in the absence of a significant family history for neuromuscular disorders. Notably, we identified involvement of the psoas muscle on MRI and muscle CT, a feature not previously documented. Proband's muscle biopsy showed an increase of ZASP expression by western blotting. Muscle fibres morphological features included peculiar sarcolemmal invaginations, pathological aggregates positive to ZASP, ubiquitin, p62 and LC3 antibodies, and the accumulation of autophagic vacuoles, suggesting that protein aggregate formation and autophagy are involved in this additional case of zaspopathy.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{%a1:%Y__493,

title = {Small Extracellular Vesicles from adipose derived stromal cells significantly attenuate in vitro the NF-kappaB dependent inflammatory/catabolic environment of osteoarthritis. },

author = {Cavallo C and Merli G and Borzì RM and Zini N and D'Adamo S and Guescini M and Grigolo B and Di Martino A and Santi S and Filardo G},

url = {https://www.nature.com/articles/s41598-020-80032-7},

doi = {10.1038/s41598-020-80032-7},

year  = {2021},

date = {2021-03-09},

journal = {Scientific Reports},

volume = {11},

number = {1},

pages = {1053},

abstract = {The therapeutic ability of Mesenchymal Stem/Stromal Cells to address osteoarthritis (OA) is mainly related to the secretion of biologically active factors, which can be found within their secreted Extracellular Vesicles including small Extracellular Vesicles (sEV). Aim of this study was to investigate the effects of sEV from adipose derived stromal cells (ADSC) on both chondrocytes and synoviocytes, in order to gain insights into the mechanisms modulating the inflammatory/catabolic OA environment. sEV, obtained by a combined precipitation and size exclusion chromatography method, were quantified and characterized, and administered to chondrocytes and synoviocytes stimulated with IL-1beta. Cellular uptake of sEV was evaluated from 1 to 12 h. Gene expression and protein release of cytokines/chemokines, catabolic and inflammatory molecules were analyzed at 4 and 15 h, when p65 nuclear translocation was investigated to study NF-kappaB pathway. This study underlined the potential of ADSC derived sEV to affect gene expression and protein release of both chondrocytes and synoviocytes, counteracting IL-1beta induced inflammatory effects, and provided insights into their mechanisms of action. sEV uptake was faster in synoviocytes, where it also elicited stronger effects, especially in terms of cytokine and chemokine modulation. The inflammatory/catabolic environment mediated by NF-kappaB pathway was significantly attenuated by sEV, which hold promise as new therapeutic strategy to address OA.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{%a1:%Y__506,

title = {TP53 drives abscopal effect by secretion of senescence-associated molecular signals in non-small cell lung cancer.},

author = {Tesei A and Arienti C and Bossi G and Santi S and {De Santis I} and Bevilacqua A and Zanoni M and Pignatta S and Cortesi M and Zamagni A and Storci G and Bonafe' M and Sarnelli A and Romeo A and Cavallo C and Bartolazzi A and Rossi S and Soriani A and Strigari L},

url = {https://jeccr.biomedcentral.com/articles/10.1186/s13046-021-01883-0},

doi = {10.1186/s13046-021-01883-0},

year  = {2021},

date = {2021-03-09},

journal = {Journal of experimental & clinical cancer research},

volume = {40},

number = {1},

pages = {89},

abstract = {Background: Recent developments in abscopal effect strongly support the use of radiotherapy for the treatment of metastatic disease. However, deeper understanding of the molecular mechanisms underlying the abscopal effect are required to best benefit a larger proportion of patients with metastasis. Several groups including ours, reported the involvement of wild-type (wt) p53 in radiation-induced abscopal effects, however very little is known on the role of wtp53 dependent molecular mechanisms. Methods: We investigated through in vivo and in vitro approaches how wtp53 orchestrates radiation-induced abscopal effects. Wtp53 bearing (A549) and p53-null (H1299) NSCLC lines were xenotransplanted in nude mice, and cultured in 2D monolayers and 3D tumor spheroids. Extracellular vesicles (EVs) were isolated from medium cell culture by ultracentrifugation protocol followed by Nanoparticle Tracking Analysis. Gene expression was evaluated by RT-Real Time, digital qRT-PCR, and dot blot technique. Protein levels were determined by immunohistochemistry, confocal anlysis, western blot techniques, and immunoassay. Results: We demonstrated that single high-dose irradiation (20 Gy) induces significant tumor growth inhibition in contralateral non-irradiated (NIR) A549 xenograft tumors but not in NIR p53-null H1299 or p53-silenced A549 (A549sh/p53) xenografts. We further demonstrates that irradiation of A549 cells in vitro induces a senescence-associated secretory phenotype (SASP) producing extracellular vesicles (EVs) expressing CD63 and carrying DNA:RNA hybrids and LINE-1 retrotransposon. IR-A549 EVs also hamper the colony-forming capability of recipient NIR A549 cells, induce senescent phenotype, nuclear expression of DNA:RNA hybrids, and M1 macrophage polarization. Conclusions: In our models, we demonstrate that high radiation dose in wtp53 tumors induce the onset of SASP and secretion of CD63+ EVs loaded with DNA:RNA hybrids and LINE-1 retrotransposons that convey senescence messages out of the irradiation field triggering abscopal effect in NIR tumors.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{%a1:%Y_440,

title = {A Reliable Flow-Based Method for the Accurate Measure of Mass Density, Size and Weight of Live 3D Tumor Spheroids},

author = {Cristaldi DA and Sargenti A and Bonetti S and Musmeci F and Delprete C and Bacchi F and Pasqua S and Cavallo C and Bonsi L and Alviano F and Gazzola D and Santi S},

url = {https://www.mdpi.com/2072-666X/11/5/465},

doi = {10.3390/mi11050465},

year  = {2020},

date = {2020-01-01},

journal = {Micromachines (Basel)},

volume = {11},

number = {5},

pages = {E465},

abstract = {Gathering precise information on mass density, size and weight of cells or cell aggregates, is crucial for applications in many biomedical fields with a specific focus on cancer research. Although few technical solutions have been presented for single-cell analysis, literature does not cover this aspect for 3D models such as spheroids. Since the research interest on such samples is notably rising, here we describe a flow-apparatus, and the associated physical method and operative protocol for the accurate measurements of mass density, size and weight. The technique is based on the detection of the terminal velocity of a free-falling sample into a specifically conceived analysis flow-channel. Moreover, in order to demonstrate the accuracy and precision of the presented flow-device, analyses were initially carried out on standardized polystyrene beads. Finally, to display the application of the proposed system for biological samples, mass density, size and weight of live SW620 tumor spheroids were analyzed. The combined measurements of such parameters can represent a step toward a deeper understanding of 3D culture models.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{%a1:%Y_475,

title = {PCAF Involvement in Lamin A/C-HDAC2 Interplay during the Early Phase of Muscle Differentiation},

author = {Santi S and Cenni V and Capanni C and Lattanzi G and Mattioli E},

url = {https://www.mdpi.com/2073-4409/9/7/1735},

doi = {10.3390/cells9071735},

year  = {2020},

date = {2020-01-01},

urldate = {2020-01-01},

journal = {Cells},

volume = {9},

number = {7},

pages = {E1735},

abstract = {Lamin A/C has been implicated in the epigenetic regulation of muscle gene expression through dynamic interaction with chromatin domains and epigenetic enzymes. We previously showed that lamin A/C interacts with histone deacetylase 2 (HDAC2). In this study, we deepened the relevance and regulation of lamin A/C-HDAC2 interaction in human muscle cells. We present evidence that HDAC2 binding to lamina A/C is related to HDAC2 acetylation on lysine 75 and expression of p300-CBP associated factor (PCAF), an acetyltransferase known to acetylate HDAC2. Our findings show that lamin A and farnesylated prelamin A promote PCAF recruitment to the nuclear lamina and lamin A/C binding in human myoblasts committed to myogenic differentiation, while protein interaction is decreased in differentiating myotubes. Interestingly, PCAF translocation to the nuclear envelope, as well as lamin A/C-PCAF interaction, are reduced by transient expression of lamin A mutated forms causing Emery Dreifuss muscular dystrophy. Consistent with this observation, lamin A/C interaction with both PCAF and HDAC2 is significantly reduced in Emery-Dreifuss muscular dystrophy myoblasts. Overall, these results support the view that, by recruiting PCAF and HDAC2 in a molecular platform, lamin A/C might contribute to regulate their epigenetic activity required in the early phase of muscle differentiation.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{%a1:%Y_459,

title = {Polyamine supplementation reduces DNA damage in adipose stem cells cultured in 3-D. },

author = {Minguzzi M and Guidotti S and Platano D and D'Adamo S and Cetrullo S and Assirelli E and Santi S and Mariani E and Trisolino G and Filardo G and Flamigni F and BorzI' RM},

url = {https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6776621/},

doi = {10.1038/s41598-019-50543-z},

year  = {2020},

date = {2020-01-01},

journal = {Scientific reports},

volume = {9},

number = {1},

pages = {14269},

abstract = {According to previous research, natural polyamines exert a role in regulating cell committment and differentiation from stemness during skeletal development. In order to assess whether distinct polyamine patterns are associated with different skeletal cell types, primary cultures of stem cells, chondrocytes or osteoblasts were dedicated for HPLC analysis of intracellular polyamines. Spermine (SPM) and Spermidine (SPD) levels were higher in adipose derived stem cells (ASC) compared to mature skeletal cells, i.e. chondrocytes and osteoblasts, confirming the connection of polyamine content with stemness. To establish whether polyamines can protect ASC against oxidative DNA damage in a 3-D differentiation model, the level of γH2AX was measured by western blot, and found to correlate with age and BMI of patients. Addition of either polyamine to ASC was able to hinder DNA damage in the low micromolecular range, with marked reduction of γH2AX level at 10 µM SPM and 5 µM SPD. Molecular analysis of the mechanisms that might underlie the protective effect of polyamine supplementation evidences a possible involvement of autophagy. Altogether, these results support the idea that polyamines are able to manage both stem cell differentiation and cell oxidative damage, and therefore represent appealing tools for regenerative and cell based applications.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{%a1:%Y_442,

title = {Spermidine Rescues the Deregulated Autophagic Response to Oxidative Stress of Osteoarthritic Chondrocytes},

author = {{D'Adamo S} and Cetrullo S and Guidotti S and Silvestri Y and Minguzzi M and Santi S and Cattini L and Filardo G and Flamigni F and Borzi' RM},

url = {https://www.sciencedirect.com/science/article/abs/pii/S0891584920303361?via%3Dihub},

doi = {10.1016/j.freeradbiomed.2020.03.029},

year  = {2020},

date = {2020-01-01},

journal = {Free radical biology & medicine},

volume = {153},

pages = {159-172},

abstract = {Oxidative stress (OS) contributes to Osteoarthritis (OA) pathogenesis and its effects are worsened by the impairment of homeostatic mechanisms such as autophagy in OA chondrocytes. Rescue of an efficient autophagic flux could therefore reduce the bulk of damaged molecules, and at the same time improve cell function and viability. As a promising dietary or intra-articular supplement to rescue autophagy in OA chondrocytes, we tested spermidine (SPD), known to induce autophagy and to reduce OS in several other cellular models. Chondrocytes were obtained from OA cartilage and seeded at high-density to keep their differentiated phenotype. The damaging effects of OS and the chondroprotective activity of SPD were assessed by evaluating the extent of cell death, oxidative DNA damage and caspase 3 activation. The autophagy promoting activity of SPD was evaluated by assessing pivotal autophagic effectors, i.e. Beclin-1 (BECN-1), microtubule-associated protein 1 light chain 3 II (LC3-II) and p62. BECN-1 protein expression was significantly increased by SPD and reduced by H2O2 treatment. SPD also rescued the impaired autophagic flux consequent to H2O2 exposure by increasing mRNA and protein expression of LC3-II and p62. SPD induction of mitophagy was revealed by immunofluorescent co-localization of LC3-II and TOM20. The key protective role of autophagy was confirmed by the loss of SPD chondroprotection upon autophagy-related gene 5 (ATG5) silencing. Significant SPD tuning of the H2O2-dependent induction of degradative (MMP-13), inflammatory (iNOS, COX-2) and hypertrophy markers (RUNX2 and VEGF) was revealed by Real Time PCR and pointed at the SPD ability of reducing NF-κB activation through autophagy induction. Conversely, blockage of autophagy led to parallel increases of oxidative markers and p65 nuclear translocation. SPD also increased the proliferation of slow-proliferating primary cultures. Taken together, our findings highlight the chondroprotective, anti-oxidant and anti-inflammatory activity of SPD and suggest that the protection afforded by SPD against OS is exerted through the rescue of the autophagic flux.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{%a1:%Y_94,

title = {Targeting Wnt/beta-catenin and PI3K/Akt/mTOR pathways in T-cell acute lymphoblastic leukemia.},

author = {Evangelisti C and Chiarini F and Cappellini A and Paganelli F and Fini M and Santi S and Martelli AM and Neri LM and Evangelisti C},

url = {https://onlinelibrary.wiley.com/doi/full/10.1002/jcp.29429},

doi = {10.1002/jcp.29429},

year  = {2020},

date = {2020-01-01},

journal = {Journal of cellular physiology},

volume = {235},

number = {6},

pages = {5413-5428},

abstract = {T-cell acute lymphoblastic leukemia (T-ALL) is an aggressive hematological disorder that results from the clonal transformation of T-cell precursors. Phosphatidylinositol 3-kinase (PI3K)/Akt/mechanistic target of rapamycin (mTOR) and canonical Wnt/beta-catenin signaling pathways play a crucial role in T-cell development and in self-renewal of healthy and leukemic stem cells. Notably, beta-catenin is a transcriptional regulator of several genes involved in cancer cell proliferation and survival. In this way, aberrations of components belonging to the aforementioned networks contribute to T-ALL pathogenesis. For this reason, inhibition of both pathways could represent an innovative strategy in this hematological malignancy. Here, we show that combined targeting of Wnt/β-catenin pathway through ICG-001, a CBP/beta-catenin transcription inhibitor, and of the PI3K/Akt/mTOR axis through ZSTK-474, a PI3K inhibitor, downregulated proliferation, survival, and clonogenic activity of T-ALL cells. ICG-001 and ZSTK-474 displayed cytotoxic effects, and, when combined together, induced a significant increase in apoptotic cells. This induction of apoptosis was associated with the downregulation of Wnt/beta-catenin and PI3K/Akt/mTOR pathways. All these findings were confirmed under hypoxic conditions that mimic the bone marrow niche where leukemic stem cells are believed to reside. Taken together, our findings highlight potentially promising treatment consisting of cotargeting Wnt/beta-catenin and PI3K/Akt/mTOR pathways in T-ALL settings. Copyrights 2020 Wiley Periodicals, Inc.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{%a1:%Y_423,

title = {Tendon Extracellular Matrix Remodeling and Defective Cell Polarization in the Presence of Collagen VI Mutations.},

author = {Antoniel M and Traina F and Merlini L and Andrenacci D and Tigani D and Santi S and Cenni V and Sabatelli P and Faldini C and Squarzoni S},

url = {https://www.mdpi.com/2073-4409/9/2/409},

doi = {10.3390/cells9020409},

year  = {2020},

date = {2020-01-01},

journal = {Cells},

volume = {9},

number = {2},

pages = {e409},

abstract = {Mutations in collagen VI genes cause two major clinical myopathies, Bethlem myopathy (BM) and Ullrich congenital muscular dystrophy (UCMD), and the rarer myosclerosis myopathy. In addition to congenital muscle weakness, patients affected by collagen VI-related myopathies show axial and proximal joint contractures, and distal joint hypermobility, which suggest the involvement of tendon function. To gain further insight into the role of collagen VI in human tendon structure and function, we performed ultrastructural, biochemical, and RT-PCR analysis on tendon biopsies and on cell cultures derived from two patients affected with BM and UCMD. In vitro studies revealed striking alterations in the collagen VI network, associated with disruption of the collagen VI-NG2 (Collagen VI-neural/glial antigen 2) axis and defects in cell polarization and migration. The organization of extracellular matrix (ECM) components, as regards collagens I and XII, was also affected, along with an increase in the active form of metalloproteinase 2 (MMP2). In agreement with the in vitro alterations, tendon biopsies from collagen VI-related myopathy patients displayed striking changes in collagen fibril morphology and cell death. These data point to a critical role of collagen VI in tendon matrix organization and cell behavior. The remodeling of the tendon matrix may contribute to the muscle dysfunction observed in BM and UCMD patients.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{%a1:%Y_439,

title = {Transportin 3 (TNPO3) and related proteins in limb girdle muscular dystrophy D2 muscle biopsies: A morphological study and pathogenetic hypothesis},

author = {Costa R and Rodia MT and Vianello S and Santi S and Lattanzi G and Angelini C and Pegoraro E and Cenacchi G},

url = {https://www.nmd-journal.com/article/S0960-8966(20)30122-X/pdf},

doi = {10.1016/j.nmd.2020.05.006},

year  = {2020},

date = {2020-01-01},

journal = {Neuromuscular disorders},

abstract = {LGMD D2 is a disease caused by TNPO3 mutation. We describe the expression of TNPO3 and selected proteins, likely modified by TNPO3 mutation, in muscle biopsies of affected patients. We also aim to find other genes involved in pathways correlated to TNPO3. Our morphological study on LGMD D2 muscle described the expression of TNPO3 and SRSF1, a splicing factor transported by TNPO3. Moreover, we investigated some sarcomeric and nuclear proteins, likely altered by TNPO3 mutation. Through an in silico approach we tried to identify genes involved in pathways that include, besides TNPO3 and SRSF1, p62 and Murf-1, altered in LGMD D2. In patients' muscles TNPO3 appeared weaker and randomly organized, with sporadic cytoplasmic aggregates positive for TNPO3; both SRSF1 and sarcomeric alpha actinin showed a different expression, while there were no alterations in the expression of the nuclear proteins. The in silico study lead to identify five genes, all coding for proteins responsible for muscle contraction. Our data suggest a possible interference in the morphology and function of myofibrillar network by mutated TNPO3; these findings are supported by the in silico identification of genes involved in muscle contraction that could help to explain the pathogenic mechanisms of LGMD D2.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{%a1:%Y_60,

title = {Genomic stability, anti-inflammatory phenotype, and up-regulation of the RNAseH2 in cells from centenarians.},

author = {Storci G and De Carolis S and Papi A and Bacalini MG and Gensous N and Marasco E and Tesei A and Fabbri F and Arienti C and Zanoni M and Sarnelli A and Santi S and Olivieri F and Mensa' E and Latini S and Ferracin M and Salvioli S and Garagnani P and Franceschi C and Bonafe' M},

url = {www.nature.com/articles/s41418-018-0255-8},

doi = {10.1038/s41418-018-0255-8},

year  = {2019},

date = {2019-09-29},

journal = {Cell death and differentiation},

volume = {26},

number = {9},

pages = {1845-1858},

abstract = {Current literature agrees on the notion that efficient DNA repair favors longevity across evolution. The DNA damage response machinery activates inflammation and type I interferon signaling. Both pathways play an acknowledged role in the pathogenesis of a variety of age-related diseases and are expected to be detrimental for human longevity. Here, we report on the anti-inflammatory molecular make-up of centenarian’s fibroblasts (low levels of IL-6, type 1 interferon beta, and pro-inflammatory microRNAs), which is coupled with low level of DNA damage (measured by comet assay and histone-2AX activation) and preserved telomere length. In the same cells, high levels of the RNAseH2C enzyme subunit and low amounts of RNAseH2 substrates, i.e. cytoplasmic RNA:DNA hybrids are present. Moreover, RNAseH2C locus is hypo-methylated and RNAseH2C knock-down up-regulates IL-6 and type 1 interferon beta in centenarian’s fibroblasts. Interestingly, RNAseH2C locus is hyper-methylated in vitro senescent cells and in tissues from atherosclerotic plaques and breast tumors. Finally, extracellular vesicles from centenarian’s cells up-regulate RNAseH2C expression and dampen the pro-inflammatory phenotype of fibroblasts, myeloid, and cancer cells. These data suggest that centenarians are endowed with restrained DNA damage-induced inflammatory response, that may facilitate their escape from the deleterious effects of age-related chronic inflammation.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{%a1:%Y_69,

title = {Polyamine supplementation reduces DNA damage in adipose stem cells cultured in 3-D.},

author = {Minguzzi M and Guidotti S and Platano D and D'Adamo S and Cetrullo S and Assirelli E and Santi S and Mariani E and Trisolino G and Filardo G and Flamigni F and Borzì RM},

url = {https://www.nature.com/articles/s41598-019-50543-z},

doi = {10.1038/s41598-019-50543-z},

year  = {2019},

date = {2019-10-03},

journal = {Scientific reports},

volume = {9},

number = {1},

pages = {14269},

abstract = {According to previous research, natural polyamines exert a role in regulating cell committment and differentiation from stemness during skeletal development. In order to assess whether distinct polyamine patterns are associated with different skeletal cell types, primary cultures of stem cells, chondrocytes or osteoblasts were dedicated for HPLC analysis of intracellular polyamines. Spermine (SPM) and Spermidine (SPD) levels were higher in adipose derived stem cells (ASC) compared to mature skeletal cells, i.e. chondrocytes and osteoblasts, confirming the connection of polyamine content with stemness. To establish whether polyamines can protect ASC against oxidative DNA damage in a 3-D differentiation model, the level of γH2AX was measured by western blot, and found to correlate with age and BMI of patients. Addition of either polyamine to ASC was able to hinder DNA damage in the low micromolecular range, with marked reduction of γH2AX level at 10 µM SPM and 5 µM SPD. Molecular analysis of the mechanisms that might underlie the protective effect of polyamine supplementation evidences a possible involvement of autophagy. Altogether, these results support the idea that polyamines are able to manage both stem cell differentiation and cell oxidative damage, and therefore represent appealing tools for regenerative and cell based applications},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{%a1:%Y_78,

title = {The N-Acetyl Phenylalanine Glucosamine Derivative Attenuates the Inflammatory/Catabolic Environment in a Chondrocyte-Synoviocyte Co-Culture System.},

author = {Pagani S and Minguzzi M and Sicuro L and Veronesi F and Santi S and Scotto D'Abusco A and Fini M and Borzì RM},

url = {https://www.nature.com/articles/s41598-019-49188-9},

doi = {10.1038/s41598-019-49188-9},

year  = {2019},

date = {2019-09-19},

journal = {Scientific reports},

volume = {9},

number = {1},

pages = {13603},

abstract = {Osteoarthritis (OA), the most prevalent degenerative joint disease, still lacks a true disease-modifying therapy. The involvement of the NF-κB pathway and its upstream activating kinases in OA pathogenesis has been recognized for many years. The ability of the N-acetyl phenylalanine glucosamine derivative (NAPA) to increase anabolism and reduce catabolism via inhibition of IKKα kinase has been previously observed in vitro and in vivo. The present study aims to confirm the chondroprotective effects of NAPA in an in vitro model of joint OA established with primary cells, respecting both the crosstalk between chondrocytes and synoviocytes and their phenotypes. This model satisfactorily reproduces some features of the previously investigated DMM model, such as the prominent induction of ADAMTS-5 upon inflammatory stimulation. Both gene and protein expression analysis indicated the ability of NAPA to counteract key cartilage catabolic enzymes (ADAMTS-5) and effectors (MCP-1). Molecular analysis showed the ability of NAPA to reduce IKKα nuclear translocation and H3Ser10 phosphorylation, thus inhibiting IKKα transactivation of NF-κB signalling, a pivotal step in the NF-κB-dependent gene expression of some of its targets. In conclusion, our data confirm that NAPA could truly act as a disease-modifying drug in OA.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{%a1:%Y_142,

title = {A Novel Role for the Interleukin-1 Receptor Axis in Resistance to Anti-EGFR Therapy.},

author = {Gelfo V and Mazzeschi M and Grilli G and Lindzen M and Santi S and D'Uva G and Gyorffy B and Ardizzoni A and Yarden Y and Lauriola M},

url = {https://www.mdpi.com/2072-6694/10/10/355 },

doi = {10.3390/cancers10100355.},

year  = {2018},

date = {2018-02-15},

journal = {Cancers},

volume = {10},

number = {10},

pages = {pii: E365},

abstract = {Cetuximab (CTX) is a monoclonal antibody targeting the epidermal growth factor receptor (EGFR), commonly used to treat patients with metastatic colorectal cancer (mCRC). Unfortunately, objective remissions occur only in a minority of patients and are of short duration, with a population of cells surviving the treatment and eventually enabling CTX resistance. Our previous study on CRC xenopatients associated poor response to CTX with increased abundance of a set of pro-inflammatory cytokines, including the interleukins IL-1A, IL-1B and IL-8. Stemming from these observations, our current work aimed to assess the role of IL-1 pathway activity in CTX resistance. We employed a recombinant decoy TRAP IL-1, a soluble protein combining the human immunoglobulin Fc portion linked to the extracellular region of the IL-1-receptor (IL-1R1), able to sequester IL-1 directly from the medium. We generated stable clones expressing and secreting a functional TRAP IL-1 into the culture medium. Our results show that IL-1R1 inhibition leads to a decreased cell proliferation and a dampened MAPK and AKT axes. Moreover, CRC patients not responding to CTX blockage displayed higher levels of IL-1R1 than responsive subjects, and abundant IL-1R1 is predictive of survival in patient datasets specifically for the consensus molecular subtype 1 (CMS1). We conclude that IL-1R1 abundance may represent a therapeutic marker for patients who become refractory to monoclonal antibody therapy, while inhibition of IL-1R1 by TRAP IL-1 may offer a novel therapeutic strategy.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{%a1:%Y_157,

title = {Functionalized Keratin as Nanotechnology-Based Drug Delivery System for the Pharmacological Treatment of Osteosarcoma.},

author = {Martella E and Ferroni C and Guerrini A and Ballestri M and Columbaro M and Santi S and Sotgiu G and Serra M and Donati DM and Lucarelli E and Varchi G and Duchi S},

url = {https://www.mdpi.com/1422-0067/19/11/3670},

doi = {10.3390/ijms19113670},

year  = {2018},

date = {2018-11-20},

journal = {International journal of molecular sciences},

volume = {19},

number = {11},

pages = {pii: E3670},

abstract = {Osteosarcoma therapy might be moving toward nanotechnology-based drug delivery systems to reduce the cytotoxicity of antineoplastic drugs and improve their pharmacokinetics. In this paper, we present, for the first time, an extensive chemical and in vitro characterization of dual-loaded photo- and chemo-active keratin nanoparticles as a novel drug delivery system to treat osteosarcoma. The nanoparticles are prepared from high molecular weight and hydrosoluble keratin, suitably functionalized with the photosensitizer Chlorin-e6 (Ce6) and then loaded with the chemotherapeutic drug Paclitaxel (PTX). This multi-modal PTX-Ce6@Ker nanoformulation is prepared by both drug-induced aggregation and desolvation methods, and a comprehensive physicochemical characterization is performed. PTX-Ce6_Ker efficacy is tested on osteosarcoma tumor cell lines, including chemo-resistant cells, using 2D and 3D model systems. The single and combined contributions of PTX and Ce6 is evaluated, and results show that PTX retains its activity while being vehiculated through keratin. Moreover, PTX and Ce6 act in an additive manner, demonstrating that the combination of the cytostatic blockage of PTX and the oxidative damage of ROS upon light irradiation have a far superior effect compared to singularly administered PTX or Ce6. Our findings provide the proof of principle for the development of a novel, nanotechnology-based drug delivery system for the treatment of osteosarcoma.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{%a1:%Y_227,

title = {A new holistic 3D non-invasive analysis of cellular distribution and motility on fibroin-alginate microcarriers using light sheet fluorescent microscopy.},

author = {Duchi S and Piccinini F and Pierini M and Bevilacqua A and Torre ML and Lucarelli E and Santi S},

url = {http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0183336},

doi = {0.1371/journal.pone.0183336},

year  = {2017},

date = {2017-02-16},

journal = {Plos One},

volume = {12},

number = {8},

pages = {e0183336},

abstract = {Cell interaction with biomaterials is one of the keystones to developing medical devices for tissue engineering applications. Biomaterials are the scaffolds that give three-dimensional support to the cells, and are vectors that deliver the cells to the injured tissue requiring repair. Features of biomaterials can influence the behaviour of the cells and consequently the efficacy of the tissue-engineered product. The adhesion, distribution and motility of the seeded cells onto the scaffold represent key aspects, and must be evaluated in vitro during the product development, especially when the efficacy of a specific tissue-engineered product depends on viable and functional cell loading. In this work, we propose a non-invasive and non-destructive imaging analysis for investigating motility, viability and distribution of Mesenchymal Stem Cells (MSCs) on silk fibroin-based alginate microcarriers, to test the adhesion capacity of the fibroin coating onto alginate which is known to be unsuitable for cell adhesion. However, in depth characterization of the biomaterial is beyond the scope of this paper. Scaffold-loaded MSCs were stained with Calcein-AM and Ethidium homodimer-1 to detect live and dead cells, respectively, and counterstained with Hoechst to label cell nuclei. Time-lapse Light Sheet Fluorescent Microscopy (LSFM) was then used to produce three-dimensional images of the entire cells-loaded fibroin/alginate microcarriers. In order to quantitatively track the cell motility over time, we also developed an open source user friendly software tool called Fluorescent Cell Tracker in Three-Dimensions (F-Tracker3D). Combining LSFM with F-Tracker3D we were able for the first time to assess the distribution and motility of stem cells in a non-invasive, non-destructive, quantitative, and three-dimensional analysis of the entire surface of the cell-loaded scaffold. We therefore propose this imaging technique as an innovative holistic tool for monitoring cell-biomaterial interactions, and as a tool for the design, fabrication and functionalization of a scaffold as a medical device.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{%a1:%Y_233,

title = {Effect of a one-step self-etch adhesive on endogenous dentin matrix metalloproteinases.},

author = {Apolonio FM and Mazzoni A and Angeloni V and Scaffa PM and Santi S and Saboia VP and Tay FR and Pashley DH and Breschi L},

url = {http://onlinelibrary.wiley.com/doi/10.1111/eos.12337/abstract;jsessionid=518F5CAC0BCA0ADC60A80D53F30477C5.f02t03},

doi = {10.1111/eos.12337},

year  = {2017},

date = {2017-02-16},

journal = {European journal of oral sciences},

volume = {125},

number = {2},

pages = {168-172},

abstract = {Degradation of the hybrid layer created in dentin by dentin adhesives is caused by enzyme activities present within the dentin matrix that destroy unprotected collagen fibrils. The aim of the present study was to evaluate the effect of a one-step self-etch adhesive system on dentinal matrix metalloproteinases 2 and 4 (MMP-2 and MMP-9, respectively) using in situ zymography and an enzymatic activity assay. The null hypothesis tested was that there are no differences in the activities of dentinal MMPs before and after treatment with a one-step adhesive system. The MMP-2 and MMP-9 activities in dentin treated with the one-step adhesive, Adper Easy Bond, were quantified using an enzymatic activity assay system. The MMP activities within the hybrid layer created by the one-step adhesive tested were also evaluated using in situ zymography. The enzymatic assay revealed an increase in MMP-2 and MMP-9 activities after treatment with adhesive. In situ zymography indicated that gelatinolytic activity is present within the hybrid layer created with the one-step self-etch adhesive. The host-derived gelatinases were localized within the hybrid layer and remained active after the bonding procedure. It is concluded that the one-step self-etch adhesive investigated activates endogenous MMP-2 and MMP-9 with the dentin matrix, which may cause collagen degradation over time.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{%a1:%Y_215,

title = {Glycogen Synthase Kinase-3beta Inhibition Links Mitochondrial Dysfunction, Extracellular Matrix Remodelling and Terminal Differentiation in Chondrocytes.},

author = {Guidotti S and Minguzzi M and Platano D and Santi S and Trisolino G and Filardo G and Mariani E and Borzì RM},

url = {www.nature.com/articles/s41598-017-12129-5},

doi = {10.1038/s41598-017-12129-5},

year  = {2017},

date = {2017-09-21},

journal = {Scientific reports},

volume = {7},

number = {1},

pages = {12059},

abstract = {Following inflammatory stimuli, GSK3 inhibition functions as a hub with pleiotropic effects leading to cartilage degradation. However, little is known about the effects triggered by its direct inhibition as well as the effects on mitochondrial pathology, that contributes to osteoarthritis pathogenesis. To this aim we assessed the molecular mechanisms triggered by GSK3β inactivating stimuli on 3-D (micromass) cultures of human articular chondrocytes. Stimuli were delivered either at micromass seeding (long term) or after maturation (short term) to explore "late" effects on terminal differentiation or "early" mitochondrial effects, respectively. GSK3β inhibition significantly enhanced mitochondrial oxidative stress and damage and endochondral ossification based on increased nuclear translocation of Runx-2 and β-catenin, calcium deposition, cell death and enhanced remodelling of the extracellular matrix as demonstrated by the increased collagenolytic activity of supernatants, despite unmodified (MMP-1) or even reduced (MMP-13) collagenase gene/protein expression. Molecular dissection of the underlying mechanisms showed that GSK3β inhibition achieved with pharmacological/silencing strategies impacted on the control of collagenolytic activity, via both decreased inhibition (reduced TIMP-3) and increased activation (increased MMP-10 and MMP-14). To conclude, the inhibition of GSK3β enhances terminal differentiation via concerted effects on ECM and therefore its activity represents a tool to keep articular cartilage homeostasis.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{%a1:%Y_218,

title = {Human White Adipocytes Convert Into "Rainbow" Adipocytes In Vitro.},

author = {Maurizi G and Poloni A and Mattiucci D and Santi S and Maurizi A and Izzi V and Giuliani A and Mancini S and Zingaretti MC and Perugini J and Severi I and Falconi M and Vivarelli M and Rippo MR and Corvera S and Giordano A and Leoni P and Cinti S},

url = {http://onlinelibrary.wiley.com/doi/10.1002/jcp.25743/abstract},

doi = {10.1002/jcp.25743.},

year  = {2017},

date = {2017-10-28},

journal = {Journal of cellular physiology},

volume = {232},

number = {10},

pages = {2887-2899},

abstract = {White adipocytes are plastic cells able to reversibly transdifferentiate into brown adipocytes and into epithelial glandular cells under physiologic stimuli in vivo. These plastic properties could be used in future for regenerative medicine, but are incompletely explored in their details. Here, we focused on plastic properties of human mature adipocytes (MA) combining gene expression profile through microarray analysis with morphologic data obtained by electron and time lapse microscopy. Primary MA showed the classic morphology and gene expression profile of functional mature adipocytes. Notably, despite their committed status, MA expressed high levels of reprogramming genes. MA from ceiling cultures underwent transdifferentiation toward fibroblast-like cells with a well-differentiated morphology and maintaining stem cell gene signatures. The main morphologic aspect of the transdifferentiation process was the secretion of large lipid droplets and the development of organelles necessary for exocrine secretion further supported the liposecretion process. Of note, electron microscope findings suggesting liposecretion phenomena were found also in explants of human fat and rarely in vivo in fat biopsies from obese patients. In conclusion, both MA and post-liposecretion adipocytes show a well-differentiated phenotype with stem cell properties in line with the extraordinary plasticity of adipocytes in vivo. J. Cell. Physiol. 232: 2887-2899, 2017. 2016 Wiley Periodicals, Inc.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{%a1:%Y_318,

title = {3D tumor spheroid models for in vitro therapeutic screening: a systematic approach to enhance the biological relevance of data obtained.},

author = {Zanoni M and Piccinini F and Arienti C and Zamagni A and Santi S and Polico R and Bevilacqua A and Tesei A},

url = {http://www.nature.com/articles/srep19103},

doi = {10.1038/srep19103},

year  = {2016},

date = {2016-02-25},

journal = {Scientific Reports},

volume = {6},

pages = {19103},

abstract = {The potential of a spheroid tumor model composed of cells in different proliferative and metabolic states for the development of new anticancer strategies has been amply demonstrated. However, there is little or no information in the literature on the problems of reproducibility of data originating from experiments using 3D models. Our analyses, carried out using a novel open source software capable of performing an automatic image analysis of 3D tumor colonies, showed that a number of morphology parameters affect the response of large spheroids to treatment. In particular, we found that both spheroid volume and shape may be a source of variability. We also compared some commercially available viability assays specifically designed for 3D models. In conclusion, our data indicate the need for a pre-selection of tumor spheroids of homogeneous volume and shape to reduce data variability to a minimum before use in a cytotoxicity test. In addition, we identified and validated a cytotoxicity test capable of providing meaningful data on the damage induced in large tumor spheroids of up to diameter in 650 μm by different kinds of treatments.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{%a1:%Y_282,

title = {A module of inflammatory cytokines defines resistance of colorectal cancer to EGFR inhibitors.},

author = {Gelfo V and Rodia MT and Pucci M and Dall'Ora M and Santi S and Solmi R and Roth L and Lindzen M and Bonafè M and Bertotti A and Caramelli E and Lollini PL and Trusolino L and Yarden Y and D'Uva G and Lauriola M},

url = {http://www.impactjournals.com/oncotarget/index.php?journal=oncotarget&page=article&op=view&path[]=12354&pubmed-linkout=1},

doi = {10.18632/oncotarget.12354},

year  = {2016},

date = {2016-02-17},

journal = {Oncotarget},

volume = {7},

number = {44},

pages = {72167-72183},

abstract = {Epidermal Growth Factor Receptor (EGFR) activates a robust signalling network to which colon cancer tumours often become addicted. Cetuximab, one of the monoclonal antibodies targeting this pathway, is employed to treat patients with colorectal cancer. However, many patients are intrinsically refractory to this treatment, and those who respond develop secondary resistance along time. Mechanisms of cancer cell resistance include either acquisition of new mutations or non genomic activation of alternative signalling routes. In this study, we employed a colon cancer model to assess potential mechanisms driving resistance to cetuximab. Resistant cells displayed increased ability to grow in suspension as colonspheres and this phenotype was associated with poorly organized structures. Factors secreted from resistant cells were causally involved in sustaining resistance, indeed administration to parental cells of conditioned medium collected from resistant cells was sufficient to reduce cetuximab efficacy. Among secreted factors, we report herein that a signature of inflammatory cytokines, including IL1A, IL1B and IL8, which are produced following EGFR pathway activation, was associated with the acquisition of an unresponsive phenotype to cetuximab in vitro. This signature correlated with lack of response to EGFR targeting also in patient-derived tumour xenografts. Collectively, these results highlight the contribution of inflammatory cytokines to reduced sensitivity to EGFR blockade and suggest that inhibition of this panel of cytokines in combination with cetuximab might yield an effective treatment strategy for CRC patients refractory to anti-EGFR targeting.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{%a1:%Y_307,

title = {Collagen VI-NG2 axis in human tendon fibroblasts under conditions mimicking injury response.},

author = {Sardone F and Santi S and Tagliavini F and Traina F and Merlini L and Squarzoni S and Cescon M and Wagener R and Maraldi NM and Bonaldo P and Faldini C and Sabatelli P},

url = {https://www.sciencedirect.com/science/article/pii/S0945053X16300233?via%3Dihub},

doi = {10.1016/j.matbio.2016.02.012},

year  = {2016},

date = {2016-02-17},

journal = {Matrix Biology},

volume = {55},

pages = {90-105},

abstract = {In response to injury, tendon fibroblasts are activated, migrate to the wound, and contribute to tissue repair by producing and organizing the extracellular matrix. Collagen VI is a microfibrillar collagen enriched in the pericellular matrix of tendon fibroblasts with a potential regulatory role in tendon repair mechanism. We investigated the molecular basis of the interaction between collagen VI and the cell membrane both in tissue sections and fibroblast cultures of human tendon, and analyzed the deposition of collagen VI during migration and myofibroblast trans-differentiation, two crucial events for tendon repair. Tendon fibroblast displayed a collagen VI microfibrillar network closely associated with the cell surface. Binding of collagen VI with the cell membrane was mediated by NG2 proteoglycan, as demonstrated by in vitro perturbation of collagen VI-NG2 interaction with a NG2-blocking antibody. Cultures subjected to wound healing scratch assay displayed collagen VI-NG2 complexes at the trailing edge of migrating cells, suggesting a potential role in cell migration. In fact, the addition of a NG2-blocking antibody led to an impairment of cell polarization and delay of wound closure. Similar results were obtained after in vitro perturbation of collagen VI extracellular assembly with the 3C4 anti-collagen VI antibody and in collagen VI-deficient tendon cultures of a Ullrich congenital muscular dystrophy patient carrying mutations in COL6A2 gene. Moreover, in vitro treatment with transforming growth factor beta1 (TGFbeta1) induced a dramatic reduction of NG2 expression, both at protein and mRNA transcript level, and the impairment of collagen VI association with the cell membrane. Instead, collagen VI was still detectable in the extracellular matrix in association with ED-A fibronectin and collagen I, which were strongly induced by TGFbeta1 treatment. Our findings reveal a critical role of the NG2 proteoglycan for the binding of collagen VI to the surface of tendon fibroblasts. By interacting with NG2 proteoglycan and other extracellular matrix proteins, collagen VI regulates fibroblasts behavior and the assembly of tendon matrix, thereby playing a crucial role in tendon repair. Copyright 2016 The Authors. Published by Elsevier B.V. All rights reserved.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{%a1:%Y_308,

title = {Tendon Extracellular Matrix Alterations in Ullrich Congenital Muscular Dystrophy.},

author = {Sardone F and Traina F and Bondi A and Merlini L and Santi S and Maraldi NM and Faldini C and Sabatelli P},

url = {http://journal.frontiersin.org/article/10.3389/fnagi.2016.00131/full},

doi = {10.3389/fnagi.2016.00131},

year  = {2016},

date = {2016-03-10},

journal = {Frontiers in Aging Neuroscience},

volume = {8},

pages = {131},

abstract = {Collagen VI (COLVI) is a non-fibrillar collagen expressed in skeletal muscle and most connective tissues. Mutations in COLVI genes cause two major clinical forms, Bethlem myopathy and Ullrich congenital muscular dystrophy (UCMD). In addition to congenital muscle weakness, patients affected by COLVI myopathies show axial and proximal joint contractures and distal joint hypermobility, which suggest the involvement of the tendon function. We examined a peroneal tendon biopsy and tenocyte culture of a 15-year-old patient affected by UCMD with compound heterozygous COL6A2 mutations. In patient's tendon biopsy, we found striking morphological alterations of tendon fibrils, consisting in irregular profiles and reduced mean diameter. The organization of the pericellular matrix of tenocytes, the primary site of collagen fibril assembly, was severely affected, as determined by immunoelectron microscopy, which showed an abnormal accumulation of COLVI and altered distribution of collagen I (COLI) and fibronectin (FBN). In patient's tenocyte culture, COLVI web formation and cell surface association were severely impaired; large aggregates of COLVI, which matched with COLI labeling, were frequently detected in the extracellular matrix. In addition, metalloproteinase MMP-2, an extracellular matrix-regulating enzyme, was increased in the conditioned medium of patient's tenocytes, as determined by gelatin zymography and western blot. Altogether, these data indicate that COLVI deficiency may influence the organization of UCMD tendon matrix, resulting in dysfunctional fibrillogenesis. The alterations of tendon matrix may contribute to the complex pathogenesis of COLVI related myopathies.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{%a1:%Y_306,

title = {TGF-beta1 differentially modulates the collagen VI alpha5 and alpha6 chains in human tendon cultures.},

author = {Sabatelli P and Sardone F and Traina F and Merlini L and Santi S and Wagener R and Faldini C},

url = {https://www.biolifesas.org/biolife/jbrha-2/},

year  = {2016},

date = {2016-12-02},

journal = {Journal of Biological Regulators and Homeostatic Agents},

volume = {30},

number = {4},

pages = {107-113},

abstract = {Collagen VI is a microfibrillar collagen with a potential regulatory role in tendon repair mechanism. We studied the expression of collagen VI alpha5 and alpha6 chains in normal human tendon fibroblast cultures, both under basal condition and in response to TGF-beta1, a potent regulator of tendon healing. Under basal condition, we found that the alpha5 chain was expressed, although to a lesser extent with respect to the alpha3 chain; in contrast, the alpha6 chain was absent. The treatment with TGFbeta1 induced an opposite effect on the expression of the alpha5 and alpha6 chains; in fact, while the alpha5 chain was dramatically reduced, the alpha6 chain was induced and released in the culture medium. These data indicate that collagen VI alpha5 and alpha6 chains are differentially involved in tendon matrix homeostasis. The alpha6 chain may represent a new potential biomarker for monitoring TGFbeta1-related events in tendon, as healing and fibrotic scar formation.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{%a1:%Y_317,

title = {Peri-Synaptic Glia Recycles Brain-Derived Neurotrophic Factor for LTP Stabilization and Memory Retention.},

author = {Vignoli B and Battistini G and Melani R and Blum R and Santi S and Berardi N and Canossa M},

url = {http://www.sciencedirect.com/science/article/pii/S0896627316306328},

doi = {10.1016/j.neuron.2016.09.031},

year  = {2015},

date = {2015-09-17},

journal = {Neuron},

volume = {92},

number = {4},

pages = {873-887},

abstract = {Glial cells respond to neuronal activation and release neuroactive molecules (termed "gliotransmitters") that can affect synaptic activity and modulate plasticity. In this study, we used molecular genetic tools, ultra-structural microscopy, and electrophysiology to assess the role of brain-derived neurotrophic factor (BDNF) on cortical gliotransmission in vivo. We find that glial cells recycle BDNF that was previously secreted by neurons as pro-neurotrophin following long-term potentiation (LTP)-inducing electrical stimulation. Upon BDNF glial recycling, we observed tight, temporal, highly localized TrkB phosphorylation on adjacent neurons, a process required to sustain LTP. Engagement of BDNF recycling by astrocytes represents a novel mechanism by which cortical synapses can expand BDNF action and provide synaptic changes that are relevant for the acquisition of new memories. Accordingly, mice deficient in BDNF glial recycling fail to recognize familiar from novel objects, indicating a physiological requirement for this process in memory consolidation.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}