Identification of novel synthetic lethal vulnerability in non small cell lung cancer by co targeting TMPRSS4 and DDR1

Finding novel targets in non-small cell lung cancer (NSCLC) is highly needed and identification of synthetic lethality between two genes is a new approach to target NSCLC. We previously found that TMPRSS4 promotes NSCLC growth and constitutes a prognostic biomarker. Here, through large-scale analyses across 5 public databases we identified consistent co-expression between TMPRSS4 and DDR1. Similar to TMPRSS4, DDR1 promoter was hypomethylated in NSCLC in 3 independent cohorts and hypomethylation was an independent prognostic factor of disease-free survival. Treatment with 5-azacitidine increased DDR1 levels in cell lines, suggesting an epigenetic regulation. Cells lacking TMPRSS4 were highly sensitive to the cytotoxic effect of the DDR1 inhibitor dasatinib. TMPRSS4/DDR1 double knock-down (KD) cells, but not single KD cells suffered a G0/G1 cell cycle arrest with loss of E2F1 and cyclins A and B, increased p21 levels and a larger number of cells in apoptosis. Moreover, double KD cells were highly sensitized to cisplatin, which caused massive apoptosis (~40%). In vivo studies demonstrated tumor regression in double KD-injected mice. In conclusion, we have identified a novel vulnerability in NSCLC resulting from a synthetic lethal interaction between DDR1 and TMPRSS4

Nanoparticles as carriers for the delivery of bevacizumab

Bevacizumab is a humanized monoclonal antibody that targets VEGF and its receptors to inhibit angiogenesis. It has been used in the treatment of several cancers, including breast cancer, metastatic non-small cell lung cancer and colorectal cancer (CRC), and off-label in the treatment of ocular pathologies that present neovascularization as diabetic retinopathy and age-related macular degeneration and corneal neovascularization. Although anti-angiogenic therapies, such as bevacizumab, have improved the clinical outcomes in cancer, their efficacy is limited due to the non-specific biodistribution and low tumor penetration. This results in high doses that can lead to undesirable side effects. Also, the use of bevacizumab in the treatment of corneal neovascularization as eye drops yields low bioavailability (<5%), which implies frequent dosing. One possible strategy would be to develop nanoparticles that are able to encapsulate bevacizumab. This could overcome some of the aforementioned drawbacks by: increasing the accumulation in the tumor site, reducing the number of administrations and improving patient compliance. This may be accomplished through the use of human serum albumin (HSA). Albumin nanoparticles have been studied as drug delivery systems because they are biocompatible, biodegradable, non-toxic and they can incorporate a large variety of drugs. As a first approach, bevacizumab-loaded albumin nanoparticles (B-NP) were produced through a desolvation procedure and subsequent freeze-drying. The resulting nanoparticles were stable without any supplementary stabilization process (e.g., cross-linkage with glutraraldehyde) and presented a high payload of active antibody. In order to improve some of the capabilities of albumin nanoparticles these particles were coated with a hydrophilic polymer, polyethylen glycol 35,000 (PEG). These particles showed to develop mucoadhesive interactions with the corneal surface of the eye after topical administration. The efficacy studies of these particles in a rat model of corneal neovascularization revealed that albumin nanoparticles improved the ocular delivery of bevacizumab and its efficacy in comparison to the free antibody. Finally, the nanoparticles were evaluated in an in vivo model of colorectal cancer (CRC). The results revealed that the presence of PEG in the nanoparticles resulted in a more sustained serum levels over 'naked' nanoparticles. Also, pegylated nanoparticles were able to reach the tumor site to inhibit tumor growth, avoiding high antibody levels in the blood (which could lead to undesirable effects)

In vivo effect of bevacizumab-loaded albumin nanoparticles in the treatment of corneal neovascularization

Corneal neovascularization (CNV) is associated with different ocular pathologies, including infectious keratitis, trachoma or corneal trauma. Pharmacological treatments based on the topical application of anti-VEGF therapies have been shown to be effective in the treatment and prevention of CNV. The aim of this work was to evaluate the effect of bevacizumab-loaded albumin nanoparticles in a rat model of CNV. Bevacizumab-loaded nanoparticles, either ?naked? (B-NP) or coated with PEG 35,000 (B-NP-PEG), were administered once a day in the eyes of animals (10 μL, 4 mg/mL every 24 h) during 7 days. Bevacizumab and dexamethasone were employed as controls and administered at the same dose every 12 h. At the end of the study, the area of the eye affected by neovascularization was about 2-times lower for animals treated with B-NP than with free bevacizumab. In the study, dexamethasone did not demonstrate an inhibitory effect on CNV at the employed dose. All of these results were confirmed by histopathological analysis, which clearly showed that eyes treated with nanoparticles displayed lower levels of fibrosis, inflammation and edema. In summary, the encapsulation of bevacizumab in human serum albumin nanoparticles improved its efficacy in an animal model of CNV.Fil: Luis de Redín, Inés. Universidad de Navarra; EspañaFil: Boiero, Carolina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Unidad de Investigación y Desarrollo en Tecnología Farmacéutica. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Unidad de Investigación y Desarrollo en Tecnología Farmacéutica; ArgentinaFil: Recalde, Sergio. Universidad de Navarra; EspañaFil: Agüeros, Maite. Universidad de Navarra; EspañaFil: Allemandi, Daniel Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Unidad de Investigación y Desarrollo en Tecnología Farmacéutica. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Unidad de Investigación y Desarrollo en Tecnología Farmacéutica; ArgentinaFil: Llabot, Juan Manuel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Unidad de Investigación y Desarrollo en Tecnología Farmacéutica. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Unidad de Investigación y Desarrollo en Tecnología Farmacéutica; ArgentinaFil: García-Layana, Alfredo. Universidad de Navarra; EspañaFil: Irache, Juan M.. Universidad de Navarra; Españ

Poly(anhydride) nanoparticles containing cashew nut proteins can induce a strong Th1 and Treg immune response after oral administration

Submitted by Ana Maria Fiscina Sampaio ([email protected]) on 2018-05-10T17:17:28Z No. of bitstreams: 1 Pereira M Poly(anhydride) nanoparticles containing....pdf: 1447772 bytes, checksum: 33e8c1ff21e8bdc1a7a57d3ad03ece44 (MD5)Approved for entry into archive by Ana Maria Fiscina Sampaio ([email protected]) on 2018-05-10T17:27:05Z (GMT) No. of bitstreams: 1 Pereira M Poly(anhydride) nanoparticles containing....pdf: 1447772 bytes, checksum: 33e8c1ff21e8bdc1a7a57d3ad03ece44 (MD5)Made available in DSpace on 2018-05-10T17:27:05Z (GMT). No. of bitstreams: 1 Pereira M Poly(anhydride) nanoparticles containing....pdf: 1447772 bytes, checksum: 33e8c1ff21e8bdc1a7a57d3ad03ece44 (MD5) Previous issue date: 2018Brazilian Ministry of Science and Technology – MCTI (SisNANO/LARNano-UFPE, CNPq # 402282/2013-2) and FACEPE (APQ #0361-4.03/14).Federal University of Pernambuco. Immunopathology Keizo-Asami Laboratory. Recife, PE, BrazilUniversity of Pernambuco. Institute of Biological Sciences. Recife, PE, BrazilFederal University of Pernambuco. Immunopathology Keizo-Asami Laboratory. Recife, PE, BrazilUniversity of Navarra. Nanomedicines and Vaccines. Research Group. Pamplona, SpainFundação Oswaldo Cruz. Instituto Gonçalo Moniz. Laboratório de Parasita-Hospedeiro Interação e Epidemiologia. Salvador, BA, BrasilUniversity of Navarra. Nanomedicines and Vaccines. Research Group. Pamplona, SpainFundação Oswaldo Cruz. Instituto Gonçalo Moniz. Laboratório de Parasita-Hospedeiro Interação e Epidemiologia. Salvador, BA, BrasilUniversity of Navarra. Nanomedicines and Vaccines. Research Group. Pamplona, SpainFederal University of Pernambuco. Immunopathology Keizo-Asami Laboratory. Recife, PE, BrazilCashew nut allergy is the second most commonly reported tree nut allergy. Traditional allergen immunotherapy presents several clinical drawbacks that can be reduced by using nanoparticles-basedallergen-delivery systems, modulating the immune response towards a protective one. In this context, the goal of this work was to assess the potential of poly(anhydride) nanoparticles (NP) for cashew nut oral immunization. Cashew nut allergens-loaded nanoparticles (CNE-NP) were prepared by solvent displacement method. After nanoparticles characterization, oral immunomodulation ability was evaluated in BALB/c mice. Our results demonstrated that CNE-NP induced a higher Th1/Th2 ratio in comparison with animals immunized with free cashew nut proteins. Indeed, a decrease in splenic Th2 cytokines (IL-4, IL-5, and IL-13), and an enhancement of pro-Th1 (IL-12 and IFN-γ) and regulatory (IL-10) cytokines was observed. Furthermore, mice orally immunized with CNE-NP presented an increased expansion of CD4+ T regulatory cells, such as CD4+Foxp3+ and CD4+LAP+, in the mesenteric lymph nodes. In conclusion, oral immunization with a single dose of poly(anhydride) nanoparticles loaded with cashew nut proteins leaded to a pro-Th1 and Treg immune response. Furthermore, their immunomodulatory properties could be introduced as a new approach for management of cashew nut allergy

Human serum albumin nanoparticles for ocular delivery of bevacizumab

Bevacizumab-loaded nanoparticles (B-NP) were prepared by a desolvation process followed by freeze-drying, without any chemical, physical or enzymatic cross-linkage. Compared with typical HSA nanoparticles cross-linked with glutaraldehyde (B-NP-GLU), B-NP displayed a significantly higher mean size (310 nm vs. 180 nm) and a lower negative zeta potential (−15 mV vs. −36 mV). On the contrary, B-NP displayed a high payload of approximately 13% when measured by a specific ELISA, whereas B-NP-GLU presented a very low bevacizumab loading (0.1 μg/mg). These results could be related to the inactivation of bevacizumab after reacting with glutaraldehyde. From B-NP, bevacizumab was released following an initial burst effect, proceeded by a continuous release of bevacizumab at a rate of 6 μg/h. Cytotoxicity studies in ARPE cells were carried out at a single dose up to 72 h and with repeated doses over a 5-day period. Neither bevacizumab nor B-NP altered cell viability even when repeated doses were used. Finally, B-NP were labeled with 99mTc and administered as eye drops in rats. 99mTc-B-NP remained in the eye for at least 4 h while 99mTc-HSA was rapidly drained from the administration point. In summary, HSA nanoparticles may be an appropriate candidate for ocular delivery of bevacizumab.Fil: Luis de Redín, Inés. Universidad de Navarra; EspañaFil: Boiero, Carolina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Unidad de Investigación y Desarrollo en Tecnología Farmacéutica. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Unidad de Investigación y Desarrollo en Tecnología Farmacéutica; ArgentinaFil: Martínez-Ohárriz, María Cristina. Universidad de Navarra; EspañaFil: Agüeros, Maite. Universidad de Navarra; EspañaFil: Ramos, Rocío. Universidad de Navarra; EspañaFil: Peñuelas, Iván. Universidad de Navarra; EspañaFil: Allemandi, Daniel Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Unidad de Investigación y Desarrollo en Tecnología Farmacéutica. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Unidad de Investigación y Desarrollo en Tecnología Farmacéutica; ArgentinaFil: Llabot, Juan Manuel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Unidad de Investigación y Desarrollo en Tecnología Farmacéutica. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Unidad de Investigación y Desarrollo en Tecnología Farmacéutica; ArgentinaFil: Irache, Juan M.. Universidad de Navarra; Españ