Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2021 Apr 27;13(5):623.
doi: 10.3390/pharmaceutics13050623.

Nucleoside-Lipid-Based Nanoparticles for Phenazine Delivery: A New Therapeutic Strategy to Disrupt Hsp27-eIF4E Interaction in Castration Resistant Prostate Cancer

Hajer Ziouziou  1   2 , Clément Paris  1 , Sébastien Benizri  3 , Thi Khanh Le  1 , Claudia Andrieu  1 , Dang Tan Nguyen  1 , Ananda Appavoo  3 , David Taïeb  1   4 , Frédéric Brunel  5 , Ridha Oueslati  2 , Olivier Siri  5 , Michel Camplo  5 , Philippe Barthélémy  3 , Palma Rocchi  1
Affiliations

Nucleoside-Lipid-Based Nanoparticles for Phenazine Delivery: A New Therapeutic Strategy to Disrupt Hsp27-eIF4E Interaction in Castration Resistant Prostate Cancer

Hajer Ziouziou et al. Pharmaceutics. .

Abstract

Heat shock protein 27 (Hsp27) has an established role in tumor progression and chemo-resistance of castration-resistant prostate cancer (CRPC). Hsp27 protects eukaryotic translation initiation factor 4E (eIF4E) from degradation, thereby maintaining survival during treatment. Phenazine derivative compound #14 was demonstrated to specifically disrupt Hsp27/eIF4E interaction and significantly delay castration-resistant tumor progression in prostate cancer xenografts. In the present work, various strategies of encapsulation of phenazine #14 with either DOTAU (N-[5'-(2',3'-dioleoyl)uridine]-N',N',N'-trimethylammonium tosylate) and DOU-PEG2000 (5'-PEG2000-2',3'-dioleoyluridine) nucleolipids (NLs) were developed in order to improve its solubilization, biological activity, and bioavailability. We observed that NLs-encapsulated phenazine #14-driven Hsp27-eIF4E interaction disruption increased cytotoxic effects on castration-resistant prostate cancer cell line and inhibited tumor growth in castration-resistant prostate cancer cell xenografted mice compared to phenazine #14 and NLs alone. Phenazine #14 NL encapsulation might represent an interesting nanostrategy for CRPC therapy.

Keywords: dialkoxyphenazine; nanoformulation; nucleolipid; prostate cancer.

PubMed Disclaimer

Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
(A) Phenazine #14 formulation with DOU-PEG2000 (N-[5′-(2′,3′-dioleoyl)uridine]-N′,N′,N′-trimethylammonium tosylate)or DOTAU (5′-PEG2000-2′,3′-dioleoyluridine). (B) Schema of phenazine #14 (in green) encapsulation with the nucleolipids (not to scale). (C) TEM image showing NPDOU-PEG2000 (bar = 1 µM). (D) Dynamic light scattering experiment recorded on NPDOU-PEG2000 + phenazine 14 samples showing objects of 78.6 nm in diameter (polydispersity index of 0.222). Zeta potential of these objects, 41.2 mV (Zeta deviation 9.41). (E) Dynamic light scattering experiment recorded on NPDOTAU+ phenazine 14 samples showing objects of 74.9 nm in diameter (polydispersity index of 0.182). Zeta potential of these objects, 71.1 mV (Zeta deviation 6.37). (F) Dynamic light scattering experiment recorded on NPDOTAU samples showing objects of 83.8 nm in diameter (polydispersity index of 0.257). Zeta potential of these objects, 67.5 mV (Zeta deviation 6.88).
Figure 2
Figure 2
The encapsulated phenazine #14 with DOTAU and DOU-PEG2000 at 100 µM inhibits the interaction of eIF4E/Hsp27. (A) Co-immunoprecipitation of eIF4E showing the interaction of eIF4E and Hsp27, (B) Western Blot analysis showing the expression levels of eIF4E and Hsp27 compared to vinculin.
Figure 3
Figure 3
Confocal microscopic distribution of phenazine #14, NPDOU-PEG2000-phenazine #14, and NPDOTAU-phenazine #14. PC-3 cells were treated at 100 µM with phenazine #14 (last panel), NPDOTAU-phenazine #14 (right panel), and NPDOU-PEG2000-phenazine #14 (left panel) with DMSO (upper right panel) as control. Auto-fluorescence of phenazine #14, NPDOTAU-phenazine #14, and NPDOU-PEG2000-phenazine #14 (green) and staining of the nucleus by DAPI (4′,6-diamidino-2-phenylindole, blue) was observed. PC-3 cells were treated with DMSO (control) during 48 h and proteins were extracted.
Figure 4
Figure 4
NPDOTAU-phenazine #14 inhibits cell viability and increases apoptosis of PC-3 cells in vitro. (A) MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide) quantification of PC-3 cell viability was performed on cells treated with NPDOTAU-phenazine #14 and NPDOU-PEG2000-phenazine #14 at different concentrations (25, 50, 100), and NT (control) during 48 h. *, p ≤ 0.1, **, p ≤ 0.01 and ***, p ≤ 0.001. (B) Apoptotic cell quantification (SubG0 phase) by flow cytometry was performed on PC-3 cells treated with the compound NPDOTAU-phenazine #14 and NPDOU-PEG2000-phenazine #14 and NT (Non-Treated cells, control) at 100 µM during 48 h and labeled with propidium iodide. The bar graph represents cell percentage in subG0 phase. *, p ≤ 0.1, **, p ≤ 0.01 and ***, p ≤ 0.001.
Figure 5
Figure 5
NPDOTAU-phenazine #14 decreased tumor volume in vivo. (A) NPDOTAU-phenazine #14 significantly enhanced anticancer activity in tumor-xenograft mice. NOD SCID mice were treated with free phenazine #14 (1 mg/kg), DOTAU, and NPDOTAU-phenazine #14 (2 mg/kg) via i.p. administration (twice per week, n = 8). PBS group was used as control (n = 6). Tumor volume was measured twice per week. Data are shown as mean ± SEM. (B) Representative pictures of the PC-3-derived xenograft tumors harvested from mice that received i.p. compound NPDOTAU-phenazine #14 or control-PBS, phenazine #14, and DOTAU after an 8-week treatment. (C) The body weight of the mice during the treatment time. Mouse body weight was measured twice per week. The graph shows that the treatments did not have side effects on body weight of the mice. Data are shown as mean ± SEM. (D) Ki-67 IHC staining of tumor tissues to assess tumor proliferation. (E) Distribution of tissue Ki-67 immunostaining intensity (measured as average optical density) according to the tumor treated with PBS, DOTAU, phenazine #14, and NPDOTAU-phenazine #14. Data are shown as mean ± SEM.
See this image and copyright information in PMC

References

    1. Ferlay J., Soerjomataram I., Dikshit R., Eser S., Mathers C., Rebelo M., Parkin D.M., Forman D., Bray F. Cancer Incidence and Mortality Worldwide: Sources, Methods and Major Patterns in GLOBOCAN 2012. Int. J. Cancer. 2015;136:E359–E386. doi: 10.1002/ijc.29210. - DOI - PubMed
    1. Tannock I.F., Wit R., Berry W.R., Horti J., Pluzanska A., Chi K.N., Oudard S., Théodore C., James N.D., Turesson I., et al. Docetaxel plus Prednisone or Mitoxantrone plus Prednisone for Advanced Prostate Cancer. N. Engl. J. Med. 2004;351:1502–1512. doi: 10.1056/NEJMoa040720. - DOI - PubMed
    1. Petrylak D.P., Tangen C.M., Hussain M.H.A., Lara P.N., Jones J.A., Taplin M.E., Burch P.A., Berry D., Moinpour C., Kohli M., et al. Docetaxel and Estramustine Compared with Mitoxantrone and Prednisone for Advanced Refractory Prostate Cancer. N. Engl. J. Med. 2004;351:1513–1520. doi: 10.1056/NEJMoa041318. - DOI - PubMed
    1. Sweeney C.J., Chen Y.-H., Carducci M., Liu G., Jarrard D.F., Eisenberger M., Wong Y.-N., Hahn N., Kohli M., Cooney M.M., et al. Chemohormonal Therapy in Metastatic Hormone-Sensitive Prostate Cancer. N. Engl. J. Med. 2015;373:737–746. doi: 10.1056/NEJMoa1503747. - DOI - PMC - PubMed
    1. James N.D., Sydes M.R., Clarke N.W., Mason M.D., Dearnaley D.P., Spears M.R., Ritchie A.W.S., Parker C.C., Russell J.M., Attard G., et al. Addition of Docetaxel, Zoledronic Acid, or Both to First-Line Long-Term Hormone Therapy in Prostate Cancer (STAMPEDE): Survival Results from an Adaptive, Multiarm, Multistage, Platform Randomised Controlled Trial. Lancet. 2016;387:1163–1177. doi: 10.1016/S0140-6736(15)01037-5. - DOI - PMC - PubMed

LinkOut - more resources