anna laurenzana - Academia.edu (original) (raw)

Papers by anna laurenzana

Rheumatology, 2021

Objectives SSc is an autoimmune disease characterized by peripheral vasculopathy and skin and int... more Objectives SSc is an autoimmune disease characterized by peripheral vasculopathy and skin and internal organ fibrosis. Accumulating evidence underlines a close association between a metabolic reprogramming of activated fibroblasts and fibrosis. This prompted us to determine the metabolism of SSc dermal fibroblasts and the effect on the vasculopathy characterizing the disease. Methods A Seahorse XF96 Extracellular Flux Analyzer was used to evaluate SSc fibroblast metabolism. In vitro invasion and capillary morphogenesis assays were used to determine the angiogenic ability of endothelial cells (ECs). Immunofluorescence, flow cytometry and real-time PCR techniques provided evidence of the molecular mechanism behind the impaired vascularization that characterizes SSc patients. Results SSc fibroblasts, compared with controls, showed a boosted glycolytic metabolism with increased lactic acid release and subsequent extracellular acidification that in turn was found to impair EC invasion an...

An alternative approach to classical surface plasmon resonance spectroscopy is dielectric-loaded ... more An alternative approach to classical surface plasmon resonance spectroscopy is dielectric-loaded waveguide (DLWG) spectroscopy, widely used in the past decades to investigate bio-interaction kinetics. Despite their wide application, a successful and clear approach to use the DLWGs for the one-step simultaneous determination of both the thickness and refractive index of organic thin films is absent in the literature. We propose here, for the first time, an experimental protocol based on the multimodal nature of DLWGs to be followed in order to evaluate the optical constants and thickness of transparent thin films with a unique measurement. The proposed method is general and can be applied to every class of transparent organic materials, with a resolution and accuracy which depend on the nature of the external medium (gaseous or liquid), the geometrical characteristics of the DLWG, and the values of both the thickness and dielectric constant of the thin film. From the experimental point of view, the method is demonstrated in a nitrogen environment with an accuracy of about 3%, for the special case of electroluminescent thin films of Eu3+β-diketonate complexes, with an average thickness of about 20 nm. The high value of the refractive index measured for the thin film with the Eu(btfa)3(t-bpete) complex was confirmed by the use of a spectroscopic model based on the Judd-Ofelt theory, in which the magnetic dipole transition 5D0 → 7F1 (Eu3+) for similar films containing Eu3+ complexes is taken as a reference. The DLWGs are finally applied to control the refractive index changes of the organic thin films under UVA irradiation, with potential applications in dosimetry and monitoring light-induced transformation in organic thin films.

European Journal of Immunology, 2020

How T‐helper (Th) lymphocyte subpopulations identified in synovial fluid from patients with juven... more How T‐helper (Th) lymphocyte subpopulations identified in synovial fluid from patients with juvenile idiopathic arthritis (JIA) (Th17, classic Th1, or nonclassic Th1) drive joint damage is of great interest for the possible use of biological drugs that inhibit the specific cytokines. Our objective was to clarify the role of such Th subpopulations in the pathogenesis of articular cartilage destruction by synovial fibroblasts (SFbs), and the effect of Th17 blockage in an animal model. SFbs were isolated from healthy subjects and patients with JIA, and peripheral blood Th lymphocytes subsets were obtained from healthy subjects. Fragments of human cartilage from healthy subjects in a collagen matrix containing JIA or normal SFbs grafted underskin in SCID mice were used to measure cartilage degradation under the effects of Th supernatants. JIA SFbs overexpress MMP9 and MMP2 and Th17 induce both MMPs in normal SFbs, while nonclassic Th1 upregulate urokinase plasminogen activator (uPA) activity. In vitro invasive phenotype of normal SFbs is stimulated with conditioned medium of Th17 and nonclassic‐Th1. In the in vivo “inverse wrap” model, normal SFbs stimulated with supernatants of Th17‐lymphocytes and nonclassic Th1 produced a cartilage invasion and degradation similar to JIA SFbs. Secukinumab inhibits the cartilage damage triggered by factors produced by Th17.

Cells, 2020

Urokinase Plasminogen Activator (uPA) Receptor (uPAR) is a well-known GPI-anchored three-domain m... more Urokinase Plasminogen Activator (uPA) Receptor (uPAR) is a well-known GPI-anchored three-domain membrane protein with pro-tumor roles largely shown in all the malignant tumors where it is over-expressed. Here we have exploited the Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/Cas9 gene knock out approach to investigate its role in the oxidative metabolism in human melanoma and colon cancer as the consequences of its irreversible loss. Knocking out PLAUR, a uPAR-encoding gene, in A375p, A375M6 and HCT116, which are two human melanoma and a colon carcinoma, respectively, we have observed an increased number of mitochondria in the two melanoma cell lines, while we evidenced an immature biogenesis of mitochondria in the colon carcinoma culture. Such biological diversity is, however, reflected in a significant enhancement of the mitochondrial spare respiratory capacity, fueled by an increased expression of GLS2, and in a decreased glycolysis paired with an increased ...

Blood, 2014

BACKGROUND: The multiple myeloma (MM) represents a process in which an asymptomatic stage of mono... more BACKGROUND: The multiple myeloma (MM) represents a process in which an asymptomatic stage of monoclonal gammopathy of undetermined significance (MGUS) precedes virtually all cases of MM. It is known that tumor progression are determined by a favorable tumor microenvironment (TME) and in this scenario fibroblasts represent the principal cellular component in the TME. A particular subpopulation of fibroblasts, cancer associated fibroblasts (CAFs), has recently raised the interest of many researchers due to their active participation in tumor growth and invasion and their association with higher malignancy grade, and poor prognosis. Recent findings indicate that the urokinase plasminogen activator (u-PA), and the urokinase receptor (u-PAR) are critical in cell invasion and degradation processes. Degradation and remodeling of the surrounding tissues are crucial in the early steps of tumor progression by facilitating expansion of the tumor mass, tumor cell proliferation, migration, and i...

Journal of Functional Foods, 2019

The "senescence-associated secretory phenotype" (SASP) supports a pro-tumoral and pro-angiogenic ... more The "senescence-associated secretory phenotype" (SASP) supports a pro-tumoral and pro-angiogenic microenvironment through increased secretion of inflammatory and growth factors. We investigated oleuropein aglycone (OLE, 10 µM, 5 weeks) effect on capillary morphogenesis induced by MRC5 fibroblast SASP in mature (hMVEC) and progenitor (ECFC) endothelial cells. In senescent fibroblasts OLE reduced NFkB signaling and the expression of several SASP factors. The levels of IL-8 and VEGF were decreased in senescent fibroblast-conditioned medium collected after OLE treatment (CM senOLE) compared to that from untreated fibroblasts (CM sen). Pre-incubation with CMsen enhanced invasive activity and capillary morphogenesis in endothelial cells. This angiogenic phenotype was significantly attenuated upon pre-incubation with CM senOLE, along with reduced MMP-2, MMP-9 and uPA secretion by endothelial cells. In conclusion, OLE can modulate angiogenesis indirectly acting on local senescent fibroblasts, a new mechanism possibly contributing to the beneficial effects of the Mediterranean diet against cancer and cardiovascular diseases.

Cancer Letters, 2017

for the study at molecular and clinical level of chronic, degenerative and neoplastic diseases to... more for the study at molecular and clinical level of chronic, degenerative and neoplastic diseases to develop novel therapies (DENOTHE).

Intracellular cholesterol metabolism was reported to modulate amyloid-β (Aβ) generation in Alzhei... more Intracellular cholesterol metabolism was reported to modulate amyloid-β (Aβ) generation in Alzheimer's disease (AD). Results presented herein demonstrated that, like brain cells, cultured skin fibroblasts from AD patients contained more cholesterol esters than fibroblasts from healthy subjects. Particularly, Oil RedO , Nile Red, and filipin staining highlighted higher levels of neutral lipids which responded to inhibitors of acyl-coenzyme A:cholesterol acyl-transferase (ACAT-1), associated with an increase in free cholesterol. ACAT-1 mRNA levels increased significantly in AD fibroblasts, whereas those of sterol regulatory element binding protein-2, neutral cholesterol ester hydrolase, and ATP-binding cassette transporter member 1 were markedly down-regulated. Instead, mRNA levels of low-density lipoprotein receptor, hydroxy-methyl-glutaryl-coenzyme A reductase, caveolin-1, and amyloid-β protein precursor (AβPP) were virtually unchanged. Notably, mRNA levels of both β-site AβPPcleaving enzyme 1 (BACE1) and neprilysin were significantly down-regulated. An increase in Aβ40 and Aβ42 immunostaining and a decrease in BACE1 active form were also found in AD versus control fibroblasts. Altogether, these findings support the hypothesis that the derangement of cholesterol homeostasis is a systemic alteration involving central but also peripheral cells of AD patients, and point to cholesterol ester levels in AD fibroblasts as an additional metabolic hallmark useful in the laboratory and clinical practice.

Leukemia, 2007

Arsenic trioxide (As 2 O 3) is an effective therapy in acute promyelocytic leukemia (APL), but it... more Arsenic trioxide (As 2 O 3) is an effective therapy in acute promyelocytic leukemia (APL), but its use in other malignancies is limited by the higher concentrations required to induce apoptosis. We have reported that trolox, an analogue of a-tocopherol, increases As 2 O 3-mediated apoptosis in a variety of APL, myeloma and breast cancer cell lines, while nonmalignant cells may be protected. In the present study, we extended previous results to show that trolox increases As 2 O 3mediated apoptosis in the P388 lymphoma cell line in vitro, as evidenced by decrease of mitochondrial membrane potential and release of cytochrome c. We then sought to determine whether this combination can enhance antitumor effects while protecting normal cells in vivo. In BDF 1 mice, trolox treatment decreased As 2 O 3-induced hepatomegaly, markers of oxidative stress and hepatocellular damage. In P388 tumor-bearing mice, As 2 O 3 treatment prolonged survival, and the addition of trolox provided a further significant increase in lifespan. In addition, the combination of As 2 O 3 and trolox inhibited metastatic spread, and protected the tumor-bearing mice from As 2 O 3 liver toxicity. Our results suggest, for the first time, that trolox might prevent some of the clinical manifestations of As 2 O 3-related toxicity while increasing its pro-apoptotic capacity and clinical efficacy in hematological malignancies.

The Journals of Gerontology: Series A, 2010

Recentresearchhasfocusedonnaturalcompoundspossiblyendowedwithantiagingeffects.Resveratrolisastilb... more Recentresearchhasfocusedonnaturalcompoundspossiblyendowedwithantiagingeffects.Resveratrolisastilbene compound produced by different plants with many biologic activities, including an antiaging effect, which has been demonstratedbothinvitroineukaryoticcellsandinvivoinmice.Westudiedtheeffectofresveratrolonculturedhuman MRC5fibroblasts,awidelyusedinvitromodelinagingstudies.ThechronictreatmentofMRC5cellsuntilsenescence with5mMresveratrolinducedasmallincreaseinthetotalnumberofreplicationscompletedbytheculturesatsenescence,showedprotectiveeffectsagainstDNAoxidativedamage,andreducedsenescence-associatedincreasesinnuclear sizeandDNAcontent.AreductioninthelevelsofacetylatedformsofH3andH4histonesandp53proteinwasalso found.

Experimental Hematology, 2009

Objective. This study aimed to investigate the mechanisms of action of WEB-2170, an inverse agoni... more Objective. This study aimed to investigate the mechanisms of action of WEB-2170, an inverse agonist of platelet-activating factor receptor, capable of inducing apoptosis in human acute myelogenous leukemia (AML) cells. Material and Methods. Gene expression profiling followed by cytofluorimetric, morphologic, and biologic analyses were used to monitor WEB-2170 effects in AML cell lines (ie, NB4, KG1, NB4-MR4, THP1, and U937) and blasts from patients with different AML (M0LM5) subtypes. PTEN silencing with small interfering RNA was also performed. Results. We have demonstrated that drug-mediated cytostasis/apoptosis in NB4 cells is characterized by upregulation of cyclin G2, p21/WAF1, NIX, TNFLa, and PTEN expression, and downregulation of cyclin D2 and BCL2 expression. We observed an increase in PTEN protein accompanied by a decrease in phospho-extracellular signal-regulated kinase 2 (ERK2) and phospho-AKT, and by forkhead box O3a (FOXO3a) cytoplasmic-nuclear translocation; the mitochondrial cytochrome C release and PARP cleavage marked the late apoptotic steps. We have found that WEB-2170 triggered apoptosis in NB4, KG1, and NB4-MR4 cells where PTEN was expressed, but not in THP1 and U937 cells where PTEN was absent. Finally, we show that PTEN silencing in NB4 cells by PTEN-specific small interfering RNA resulted in a significant reduction of drug-induced apoptosis. Conclusion. We demonstrated that WEB-2170 is a powerful antileukemic agent with interesting translational opportunities to treat AML and described mechanisms of drug-induced intrinsic and extrinsic apoptosis both in AML cell lines and blasts from AML patients by addressing PTEN as the master regulator of the whole process.

Cancer Research, 2009

Transcriptional silencing via promoter methylation of genes important for cell growth and differe... more Transcriptional silencing via promoter methylation of genes important for cell growth and differentiation plays a key role in myeloid leukemogenesis. We find that clinically achievable levels of 5-aza-2′-deoxycytidine (5-AZA-dC), a potent inhibitor of DNA methylation, can modify chromatin and restore the ability of tumor necrosis factor α (TNFα) to induce monocytic differentiation of the acute myeloid leukemia cells NB4 and U937. Although 5-AZA-dC cannot fully induce differentiation, we show that 5-AZA-dC acts directly on TNFα-responsive promoters to facilitate TNFα-induced transcriptional pathways leading to differentiation. 5-AZA-dC regulates the expression of Dif-2, a TNFα target gene, by deacetylating chromatin domains in a methylation-dependent manner. Chromatin immunoprecipitation analyses of the Dif-2 promoter show histone hyperacetylation and a recruitment of the nuclear factor-κB transcription factor in response to 5-AZA-dC. Furthermore, 5-AZA-dC plus TNFα enhances the leve...

Nanomaterials, 2022

In this study, we report the realization of drug-loaded smart magnetic nanocarriers constituted b... more In this study, we report the realization of drug-loaded smart magnetic nanocarriers constituted by superparamagnetic iron oxide nanoparticles encapsulated in a dual pH- and temperature-responsive poly (N-vinylcaprolactam-co-acrylic acid) copolymer to achieve highly controlled drug release and localized magnetic hyperthermia. The magnetic core was constituted by flower-like magnetite nanoparticles with a size of 16.4 nm prepared by the polyol approach, with good saturation magnetization and a high specific absorption rate. The core was encapsulated in poly (N-vinylcaprolactam-co-acrylic acid) obtaining magnetic nanocarriers that revealed reversible hydration/dehydration transition at the acidic condition and/or at temperatures above physiological body temperature, which can be triggered by magnetic hyperthermia. The efficacy of the system was proved by loading doxorubicin with very high encapsulation efficiency (>96.0%) at neutral pH. The double pH- and temperature-responsive natu...

Journal of Biological Engineering

Journal of Investigative Dermatology

Frontiers in Oncology

uPAR is a globular protein, tethered to the cell membrane by a GPI-anchor involved in several can... more uPAR is a globular protein, tethered to the cell membrane by a GPI-anchor involved in several cancer-related properties and its overexpression commonly correlates with poor prognosis and metastasis. We investigated the consequences of uPAR irreversible loss in human melanoma and colon cancer cell lines, knocking out its expression by CRISPR/Cas9. We analyzed through flow cytometry, western blotting and qPCR, the modulation of the most known cancer stem cells-associated genes and the EGFR while we observed the proliferation rate exploiting 2D and 3D cellular models. We also generated uPAR “rescue” expression cell lines as well as we promoted the expression of only its 3’UTR to demonstrate the involvement of uPAR mRNA in tumor progression. Knocking out PLAUR, uPAR-encoding gene, we observed an inhibited growth ratio unexpectedly coupled with a significant percentage of cells acquiring a stem-like phenotype. In vivo experiments demonstrated that uPAR loss completely abrogates tumorigen...

Soft Matter

In this work, for the first time, snail slime from garden snails “Helix Aspersa Müller”, has been... more In this work, for the first time, snail slime from garden snails “Helix Aspersa Müller”, has been used to induce the formation of eco-friendly gold nanoparticles (AuNPs-SS) suitable for biomedical applications.