Grafting of anti-nucleolin aptamer into preformed and remotely loaded liposomes through aptamer-cholesterol post-insertion

Hamdi Nsairat; Ismail S Mahmoud; Fadwa Odeh; Duaa Abuarqoub; Hafsa Al-Azzawi; Rand Zaza; Malak I Qadri; Said Ismail; Abeer Al Bawab; Abdalla Awidi; Walhan Alshaer

doi:10.1039/d0ra07325c

Grafting of anti-nucleolin aptamer into preformed and remotely loaded liposomes through aptamer-cholesterol post-insertion

RSC Adv. 2020 Oct 1;10(59):36219-36229. doi: 10.1039/d0ra07325c. eCollection 2020 Sep 28.

Authors

Hamdi Nsairat^{1

2}, Ismail S Mahmoud³, Fadwa Odeh^{1

4}, Duaa Abuarqoub^{5

6}, Hafsa Al-Azzawi⁶, Rand Zaza⁶, Malak I Qadri¹, Said Ismail^{7

8}, Abeer Al Bawab^{1

4}, Abdalla Awidi^{6

7

9}, Walhan Alshaer⁶

Affiliations

¹ Department of Chemistry, The University of Jordan Amman 11942 Jordan hammdi2000@yahoo.com +962 796167327.
² Pharmacological and Diagnostic Research Center, Faculty of Pharmacy, Al-Ahliyya Amman University Amman 19328 Jordan.
³ Department of Medical Laboratory Sciences, Faculty of Allied Health Sciences, The Hashemite University Zarqa 13133 Jordan.
⁴ HMCSR, The University of Jordan Amman 11942 Jordan.
⁵ Department of Pharmacology and Biomedical Sciences, Faculty of Pharmacy and Medical Sciences, University of Petra Amman 11180 Jordan.
⁶ Cell Therapy Center, The University of Jordan Amman 11942 Jordan walhan.alshaer@ju.edu.jo +962 65355000 Ext. 23960 +962 790823678.
⁷ Faculty of Medicine, The University of Jordan Amman 11942 Jordan.
⁸ Qatar Genome Project, Qatar Foundation Doha Qatar.
⁹ Department of Hematology and Oncology, Jordan University Hospital, The University of Jordan Amman 11942 Jordan.

Abstract

A new combination strategy of an active loading and active targeting approach was applied in this work. The liposomes actively loaded with Curcumin (CRM) (Lip_CRM) were decorated with cholesterol tagged-anti-nucleolin AS1411 aptamer (NCL) via a new post-insertion approach, utilizing the cholesterol as a wedge to incorporate aptamer into the surface of the liposome bilayer. A successful NCL post-insertion was verified by agarose gel electrophoresis and dynamic light scattering (DLS). The cellular uptake of Apt_NCL-Lip was investigated using flow cytometry and Confocal Laser Scanning Microscopy (CLSM) on two different human breast cancer cell lines (MCF-7 and MDA-MB-231). The uptake and cytotoxicity of loaded CRM were investigated using flow cytometry and MTT assay. Our results showed successful post insertion of NCL aptamer to the surface of Lip. Also, higher cellular uptake was noted for Apt_NCL-Alexa-Lip_Rhod compared to blank Lip_Rhod in both cell lines. Moreover, CLSM showed prominent endocytosis and uptake of Apt_NCL-Alexa-Lip_Rhod into the cytoplasm of breast cancer cells. Furthermore, the results showed a significant increase in the uptake and cytotoxicity of Apt_NCL-Lip_CRM compared to Lip_CRM in both cell lines. Overall, our results demonstrate a successful post-insertion of cholesterol-tagged aptamer into liposomes and the possible combination between active loading and active targeting.