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
. 2018 Jun;28(2):97-105.
doi: 10.1080/08982104.2016.1274756. Epub 2017 Feb 10.

PEGylated-nanoliposomal clusterin for amyloidogenic light chain-induced endothelial dysfunction

Diana Guzman-Villanueva  1   2 , Raymond Q Migrino  3   4 , Seth Truran  3 , Nina Karamanova  3 , Daniel A Franco  3 , Camelia Burciu  3 , Subhadip Senapati  5 , Dobrin Nedelkov  6 , Parameswaran Hari  7 , Volkmar Weissig  1   2
Affiliations

PEGylated-nanoliposomal clusterin for amyloidogenic light chain-induced endothelial dysfunction

Diana Guzman-Villanueva et al. J Liposome Res. 2018 Jun.

Abstract

Light chain (AL) amyloidosis is a disease associated with significant morbidity and mortality arising from multi-organ injury induced by amyloidogenic light chain proteins (LC). There is no available treatment to reverse the toxicity of LC. We previously showed that chaperone glycoprotein clusterin (CLU) and nanoliposomes (NL), separately, restore human microvascular endothelial function impaired by LC. In this work, we aim to prepare PEGylated-nanoliposomal clusterin (NL-CLU) formulations that could allow combined benefit against LC while potentially enabling efficient delivery to microvascular tissue, and test efficacy on human arteriole endothelial function. NL-CLU was prepared by a conjugation reaction between the carboxylated surface of NL and the primary amines of the CLU protein. NL were made of phosphatidylcholine (PC), cholesterol (Chol) and 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[carboxy(polyethylene glycol)-2000] (DSPE-PEG 2000 carboxylic acid) at 70:25:5 mol%. The protective effect of NL-CLU was tested by measuring the dilation response to acetylcholine and papaverine in human adipose arterioles exposed to LC. LC treatment significantly reduced the dilation response to acetylcholine and papaverine; co-treatment of LC with PEGylated-nanoliposomal CLU or free CLU restored the dilator response. NL-CLU is a feasible and promising approach to reverse LC-induced endothelial damage.

Keywords: Amyloidosis; PEGylation; clusterin; light chain; liposomes.

PubMed Disclaimer

Conflict of interest statement

Declaration of interest

The authors report no conflicts of interest. The authors alone are responsible for the content and writing of this article. R.Q.M. was funded with the National Institutes of Health (NIA R21AG044723), Veterans Affairs Merit (I01BX007080) and the Amyloidosis Foundation.

Figures

Figure 1
Figure 1
Schematic representation of the conjugation reaction between PEG-functionalized liposomes and the protein CLU via a carboxy-to-amine reaction in the presence of the crosslinker and stabilizer EDC and NHS.
Figure 2
Figure 2
Determination of particle size distribution (A) and zeta potential (B) of nanoliposomes before and after CLU conjugation. Statistically significant differences were found between the non-PEGylated (PC:Chol) and PEGylated formulations. *p<0.05 versus PC:Chol, ***p<0.001 versus PC:Chol, respectively.
Figure 3
Figure 3
AFM imaging of nanoliposomes. (A) 2D images of CLU protein, (B) PEG-functionalized liposomes, (C) purified CLU-PEGylated nanoliposomes and (D) their height analysis. ***p<0.001 versus CLU, +++p<0.001 versus CLU-PEGylated nanoliposome.
Figure 4
Figure 4
Stability assay of PEG-functionalized nanoliposomes. The integrity of PEG-functionalized liposomes at room temperature and 37 °C was monitored by DLS after 1, 24, 48 and 168 h.
Figure 5
Figure 5
Human arteriole dilation response. A and B show response to acetylcholine with A showing maximum dilation response (10−4 M dose) and B showing EC50 values. Light chain treatment reduced dilator response to acetylcholine compared to baseline control. Co-treatment of LC with free CLU and PEGylated-nanoliposomal CLU (NL-CLU) restored dilator response to acetylcholine. Note that the y-axis of B is in inverse order. C shows response to papaverine. There was a modest reduction in dilator response in LC treated arterioles; LC co-treatment with free CLU or NL-CLU showed no significant difference with baseline control response. There was no significant difference in dilator response to acetylcholine or papaverine in arterioles treated with LC and NL-CLU versus LC and free CLU.
See this image and copyright information in PMC

Similar articles

See all similar articles

References

    1. Canet-Ferrer J, Coronado E, Forment-Aliaga A, Pinilla-Cienfuegos E. Correction of the tip convolution effects in the imaging of nanostructures studies though scanning force microscopy. Nanotechnology. 2014;25:395703. - PubMed
    1. Coelho T, Adams D, Silva A, et al. Safety and efficacy of RNAi therapy for transthyretin amyloidosis. N Engl J Med. 2013;369:819–29. - PubMed
    1. Dent DM. Conjugation methods. In: Wild D, editor. The immunoassay handbook. UK: Elsevier Ltd; 2005. pp. 213–223.
    1. Dispenzieri A, Gertz MA, Kyle RA, et al. Serum cardiac troponins and N-terminal pro-brain natriuretic peptide: a staging system for primary systemic amyloidosis. J Clin Oncol Off J Am Soc Clin Oncol. 2004a;22:3751–7. - PubMed
    1. Dispenzieri A, Gertz MA, Kyle RA, et al. Prognostication of survival using cardiac troponins and N-terminal pro-brain natriuretic peptide in patients with primary systemic amyloidosis undergoing peripheral blood stem cell transplantation. Blood. 2004b;104:1881–7. - PubMed

MeSH terms