https://hdl.handle.net/10955/34 DiBEST 2026-04-07T14:07:31Z https://hdl.handle.net/10955/5601 Livelli fisiologici della Cromogranina A esercitano potente effetto cardioprotettivo contro la cardiotossicità indotta da doxorubicina Granieri, Maria Concetta; Pasqua, Teresa; Angelone, Tommaso The clinical use of doxorubicin (Doxo), a widely used anticancer chemotherapeutic drug, is limited by dose-dependent cardiotoxicity. We have investigated whether chromogranin A (CgA), a cardioregulatory protein released in the blood by the neuroendocrine system and by the heart itself, may contribute to the regulation of cardiotoxic and antitumor activities of Doxo. We evaluated whether and to which extent, the in vivo administration of physiological doses of recombinant full-length CgA may exert cardioprotection in a Doxo-induced cardiotoxicity rat model and modulate the anticancer activity of Doxo in murine models of melanoma, fibrosarcoma, lymphoma, and lung cancer, respectively. The effect of Doxo on circulating levels of CgA was also investigated. At the end of the treatment, hearts were perfused by Langendorff method and Ischemic protocols were used. Western Blot, Immunohistochemistry techniques, and ELISA assay were used for molecular analysis and plasma measurements of TNFα, IL-1β, ROS, LDH, cTnT and CgA. We found that CgA mitigated Doxo-dependent adverse effects, as revealed by the reduction of pro-inflammatory and cardiotoxic markers. Doxo reduced CgA plasma levels. When given together with Doxo, CgA increased the systolic function after ischemia and reduced the infarct size, compared to the Doxo group alone. Molecular analysis indicated that CgA reduced CTGF expression, induced activation of the RISK and SAFE pathways and of AMPK, and reduced inflammatory targets such as iNOS, COX2, NLRP3. CgA reduced intracardiac ROS and the expression of AOX-1 and XO. CgA-dependent cardioprotection was mediated by ARC activation, by the increase of Bcl2 and the reduction of BAX, Caspase3 and the apoptotic nuclei. Furthermore, we observed that Doxo reduces the intracardiac expression and release of CgA (i.e., an important cardioprotective agent) in the blood. On the other hand, CgA did not impair the anticancer activity of Doxo in all the investigated murine models. These data suggest that CgA could mitigate Doxo-induced cardiotoxicity by limiting ischemic injury. The protein can be proposed as a possible biomarker and the administration of exogenous CgA to patients with low levels of the endogenous protein might represent a novel approach to prevent Doxo-induced adverse events without impairing its antitumor effects. UNIVERSITA’ DELLA CALABRIA Dipartimento di Biologia, Ecologia e Scienze della Terra (DiBEST) Dottorato di Ricerca in Scienze della vita Ciclo XXXII 2020-02-14T00:00:00Z https://hdl.handle.net/10955/5579 Design and biological evaluation of new anticancer agents vehicled in innovative pharmaceutical matrices Ceramella, Jessica; Cerra, Maria Carmela; Gabriele, Domenico; Sinicropi, Maria Stefania Tra tutti i tumori, quello al seno e all’utero rappresentano la principale causa di morte nelle donne. Una grande sfida è oggi rappresentata dallo sviluppo di agenti nuovi e più efficaci in grado di possedere buona attività antitumorale, evitando l’insorgenza di effetti collaterali e di fenomeni di resistenza. In questo ambito, l’attività svolta durante il dottorato di ricerca ha riguardato lo studio delle proprietà antitumorali di nuove molecole di sintesi, quali analoghi di noti antitumorali. In particolare, la ricerca ha evidenziato gli interessanti effetti antiproliferativi su diverse linee cellulari di tumore al seno a all’utero di composti a struttura carbazolica, analoghi dei farmaci noti Ellipticina e Vinblastina. Molti di questi composti sono stati in grado di indurre morte cellulare per apoptosi nelle cellule tumorali, per inibizione delle topoisomerasi e/o della polimerizzazione della tubulina. Una promettente attività multi-target si è anche ottenuta con complessi metallici dell’oro a nucleo carbenico, analoghi del Cisplatino, molecole di grande interesse e che, pertanto, potrebbero rappresentare validi candidati per lo sviluppo di nuovi agenti antitumorali. Considerando poi che negli ultimi anni il riutilizzo di farmaci già noti, in grado di agire su nuovi target, risulta di notevole interesse scientifico, in quanto ciò consente di ridurre gli sforzi della ricerca sia in termini pratici che economici, si è pensato di valutare l’attività antitumorale esercitata da alcuni analoghi sintetici della Talidomide. È risultato che essi sono stati capaci di indurre apoptosi interferendo con i processi di metastatizzazione e di angiogenesi. Ulteriori investigazioni hanno riguardato lo studio di molecole di origine naturale ed in particolare le proprietà biologiche della Quercetina e di suoi analoghi. Alcuni di essi hanno mostrato, oltre ad una marcata attività antitumorale sulla linea metastatica di breast cancer MDA-MB-231, anche una duplice azione, in quanto abili a proteggere le cellule sane dallo stress ossidativo ed anche ad indurre produzione di specie reattive dell’ossigeno, con conseguente morte cellulare nelle cellule tumorali. Inoltre, sono state realizzate e studiate alcune formulazioni per farmaci noti quali il Sunitinib, la Quercetina e il Cisplatino, che hanno permesso di ottenere molecole con profili farmaceutici migliorati, in termini di solubilità, tossicità, stabilità e rilascio. È noto, infatti, che lo sviluppo di nuovi agenti è molto dispendioso in termini di tempo e denaro, perciò oggigiorno il miglioramento della sicurezza e dell’efficacia di farmaci noti attraverso il drug delivery sta diventando un’alternativa “smart”, soprattutto in campo oncologico. I promettenti risultati ottenuti durante la ricerca, considerato che le varie molecole studiate sono state preparate con vie sintetiche semplici, che molte di esse hanno mostrato una buona attività antitumorale con assenza di citotossicità sulle cellule sane e che hanno coinvolto importanti target biologici, studi futuri verteranno sull’approfondimento delle loro proprietà farmacocinetiche e farmacodinamiche grazie a studi pre-clinici. Ciò consentirà di aprire nuove ed interessanti prospettive per lo sviluppo e l’impiego di questi “tools” in oncologia. Università della Calabria. Corso di laurea in Biologia, Ecologia e Scienze della Terra. Dottorato di ricerca in Scienze della Vita. Ciclo XXXII 2020-02-25T00:00:00Z https://hdl.handle.net/10955/5572 Monitoraggio di un gruppo di insetti predatori, i coleotteri carabidi, ai fini della conservazione e dello sviluppo sostenibile dell'agricoltura nel parco nazionale della Sila Cavaliere, Francesco; Cerra, Maria Carmela; Brandmayr, Pietro; Giglio, Anita Agricultural management practices are known to cause lethal and sub-lethal effects on animals inhabiting croplands. The persistence and residual accumulation of pesticides in the soil are detrimental to the environment and pose a risk for human health and for species providing biocontrol ecosystem services. Carabid beetles are useful bioindicators to evaluate the ecological effects of agrochemicals due to their ecological role as predators in farmland. In order to assess the sub-lethal effects at the community, species and organism levels, on non-target species caused by regular pesticide applications, an environmental monitoring was performed over a period of two years, in two sites around the agricultural area to Sila National Park. One site is a conventional farm (39°16'58.05"N, 16°38'43.26"E, 1240 m a.s.l., Torre Garga Farm, Calabria, Italy), it was treated with herbicides, pesticides and fungicides used for pest control. The second site, chosen as control, is an organic farm (39°17'10.28"N, 16°42'28.33"E, 1150 m a.s.l., Macchia di Tuono Farm, Calabria, Italy). To check the direct sublethal effect of field exposure on species abundance and density at the community level, carabid beetles were collected in vivo by pitfall traps and identified using dichotomous keys. To asses the direct sub-lethal effect of field exposure at the organism level, the body size, the constitutive immune responses and genotoxicity we quantified in adults of Harpalus rufipes, a generalist predator taxonomically and ecologically well known, widely distributed and easy to collect in crops. The findings of this study indicated that chemical treatments admitted in the conventionally managed fields have negative effects on the abundance and density of species inhabiting croplands, the low level of PO and lysozyme-like enzyme activities and the DNA damage recorded in haemocytes indicated that field treatments might influence this non-target soil-dwelling species. However, field treatments did not cause morphometric changes in the body size of adults. The reduction of humoral responses and the genotoxic effect recorded on haemocytes may result in an increased susceptibility of this species to pathogens. From an ecoimmunological point of view, a modification of other basic life history traits such as reproduction, dispersal activity and predation may occur with effects on the adult fitness, resulting in changes of the population structure and in a reduction of the biocontrol activity for pest species in agroecosystem. Università della Calabria. Dipartimento di Biologia, Ecologia e Scienze della Terra Dottorato di Ricerca in Scienze della Vita. Ciclo XXXII 2020-02-13T00:00:00Z https://hdl.handle.net/10955/5571 Molecular mechanisms involved in the regulation of certain miRNAs in breast cancer cells and cancer-associated fibroblasts Sebastiani, Anna; Maggiolinu, Marcello; Cerra, Maria Carmela MicroRNAs (miRNAs) are non-coding small RNA molecules that regulate gene expression by inhibiting the translation of target mRNAs. It has been well demonstrated that miRNAs play a regulatory role in different pathophysiological conditions, including breast cancer. In this regard, a distinct miRNA may be up regulated in certain malignancies and downregulated in others, suggesting both an oncogenic and tumor suppressor role, respectively. It has also reported that in breast cancer estrogens may regulate miRNA expression through the involvement of the classic estrogen receptor (ER) alpha and the G protein coupled receptor, GPER. By using RT-PCR and western blotting assays, as well as by gene silencing approaches, in the present work we provide novel insights on the involvement of miRNAs in the breast tumor progression. In particular, we ascertained that miR-221 expression is up-regulated in breast cancer cells SkBr3 and MDA-MB 231 and in cancer-associated fibroblasts (CAFs). In addition, we found that the proliferative and migratory effects induced by miR-221 in these cell types are mediated by interfering the A20/c-Rel/CTGF pathway. These stimulatory effects are abolished silencing the expression of the NF-κB component, c-Rel and the Connective Tissue Growth Factor (CTGF), as well as using the specific Locked Nucleic Acid (LNA)-Inhibitor of miR-221 (LNA-i-miR-221). In a second work, we evaluated the expression of 754 miRNAs by TaqMan Array in ER-negative and GPER-positive SkBr3 breast cancer cells and in CAFs upon 17-β-estradiol (E2) treatment. Among the miRNAs down-regulated by 100nM E2 for 4 hours in these cell types, we identified miR-338-3p that is positively correlated with overall survival in breast cancer patients as ascertained by METABRIC database analysis. According to these data, a miR-338-3p mimic sequence decreased and a miR-338-3p inhibitor sequence rescued the proliferative effects and cell progression induced by 100nM E2 and G-1, selective ligand of GPER in SkBr3 breast cancer cells and CAFs. Overall, these results provide new insights on the miRNA action in breast cancer, suggesting their use as an innovative therapeutic approach in this malignancy. Università della Calabria. Dipartimento di Biologia, Ecologia e Scienze della Terra Dottorato di Ricerca in Scienze della Vita. XXXII Ciclo 2020-02-03T00:00:00Z