000075428 001__ 75428
000075428 005__ 20210318091408.0
000075428 0247_ $$2doi$$a10.1016/S0016-5085(17)30503-6
000075428 0248_ $$2sideral$$a107494
000075428 037__ $$aART-2017-107494
000075428 041__ $$aeng
000075428 100__ $$aNeira, José L.
000075428 245__ $$aBlocking Nupr1 Protein, A Successful Approach for Pancreatic Adenocarcinoma Treatment
000075428 260__ $$c2017
000075428 5060_ $$aAccess copy available to the general public$$fUnrestricted
000075428 5203_ $$aBackground: Pancreatic ductal adenocarcinoma (PDAC) is the most common type of pancreatic malignancy. Nuclear protein 1 (NUPR1) is a recognized protein, over-expressed and involved in PDAC development. NUPR1 belongs to the special class of intrinsically disordered proteins (IDPs) and it is implicated in cell signaling and regulatory functions. The multifunctional nature of NUPR1 renders it as an attractive target for drug design and development.
Aim: Identify a small molecule inhibiting protein-protein interactions in NUPR1 and able to interfere with any of NUPR1 key oncogenic activities, thus, constituting a new chemotherapy strategy against PDAC.
Methods: Ligand-induced stabilization against thermal denaturation (thermal-shift assay) was employed for identifying potential NUPR1-interacting compounds. An in vitro molecular screening based on thermal denaturation of NUPR1 in the presence of a variety of potential ligands was performed using a collection of 1120 compounds. All compounds are FDA-approved drugs for different therapeutic indications, exhibiting high chemical and pharmacological diversity, as well as good bioavailability and safety parameters in humans. Direct interaction of selected compounds with NUPR1 was assessed experimentally (calorimetry, fluorescence spectroscopy, nuclear magnetic resonance, and proximity ligation assay) and computationally (molecular dynamics simulations). Compound efficacy was determined in PDAC-derived cell-based assays and in vivo assays on xenografted PDACderived cells in mice. Comparisons of treatment outcome were tested for statistical significance by using the t-test, and statistical significance was assumed at a p-value lower than 0.05.
Results: Fifteen candidates were selected, and their interactions with NUPR1 were characterized. In vitro experiments with MiaPaCa-2 cells treated using 10 µM of compounds for 6 days showed that two of the compounds (C13 and C15) were very efficient in diminishing cell viability (10 ± 3% and 26 ± 7%, respectively; assays in triplicates (n = ) p= 0.01). These values were similar to those obtained with oxaliplatin (10 ± 2%; p= 0.01). Also, they reduced cell migration (from 10-20% wound-healing ability compared to 50% in control assays; p= 0.05) and colony formation (completely suppressed in the presence of both compounds; p= 0.01). In addition, the most promising compound, C15, interfered with the interaction of NUPR1 with MSL1, one of the NUPR1 binding partners (Figure 1). The administration of a 10 mg/Kg dose of C15 promoted complete arrest of tumor development on xenografted PDAC-derived cells in mice (Figure 2).
Conclusion: We report the discovery of a compound specifically active against PDAC and interfering with NUPR1. In addition, we demonstrate that it is possible to identify small molecules able to modulate the function of complex targets such as IDPs.
000075428 540__ $$9info:eu-repo/semantics/openAccess$$aby-nc-nd$$uhttp://creativecommons.org/licenses/by-nc-nd/3.0/es/
000075428 590__ $$a20.773$$b2017
000075428 591__ $$aGASTROENTEROLOGY & HEPATOLOGY$$b1 / 80 = 0.013$$c2017$$dQ1$$eT1
000075428 592__ $$a7.958$$b2017
000075428 593__ $$aGastroenterology$$c2017$$dQ1
000075428 655_4 $$ainfo:eu-repo/semantics/article$$vinfo:eu-repo/semantics/acceptedVersion
000075428 700__ $$aBintz, Jennifer
000075428 700__ $$0(orcid)0000-0002-9336-1563$$aArruebo, Maria
000075428 700__ $$aRizzuti, Bruno
000075428 700__ $$aBonacci, Thomas
000075428 700__ $$0(orcid)0000-0002-1232-6310$$aVega Sánchez, Sonia
000075428 700__ $$aLanas, Angel
000075428 700__ $$0(orcid)0000-0001-5702-4538$$aVelázquez-Campoy, Adrian$$uUniversidad de Zaragoza
000075428 700__ $$aIovanna, Juan L.
000075428 700__ $$0(orcid)0000-0001-5664-1729$$aAbian, Olga$$uUniversidad de Zaragoza
000075428 7102_ $$11002$$2060$$aUniversidad de Zaragoza$$bDpto. Bioq.Biolog.Mol. Celular$$cÁrea Bioquímica y Biolog.Mole.
000075428 773__ $$g152, 5, Supp. 1 (2017), S42$$pGastroenterology$$tGastroenterology$$x0016-5085
000075428 8564_ $$s146806$$uhttps://zaguan.unizar.es/record/75428/files/texto_completo.pdf$$yPostprint
000075428 8564_ $$s107960$$uhttps://zaguan.unizar.es/record/75428/files/texto_completo.jpg?subformat=icon$$xicon$$yPostprint
000075428 909CO $$ooai:zaguan.unizar.es:75428$$particulos$$pdriver
000075428 951__ $$a2021-03-18-09:10:12
000075428 980__ $$aARTICLE