Characterization of CD44-Mediated Cancer Cell Uptake and Intracellular Distribution of Hyaluronan-Grafted Liposomes

Abstract

Hyaluronan (HA) is a biocompatible and biodegradable linear polysaccharide which is of interest for tumor targeting through cell surface CD44 receptors. HA binds with high affinity to CD44 receptors, which are overexpressed in many tumors and involved in cancer metastasis. In the present study, we investigated the impact of HA molecular weight (MW), grafting density, and CD44 receptor density on endocytosis of HA-grafted liposomes (HA-liposomes) by cancer cells. Additionally, the intracellular localization of the HA-liposomes was determined. HAs of different MWs (5-8, 10-12, 175-350, and 1600 kDa) were conjugated to liposomes with varying degrees of grafting density. HA surface density was quantified using the hexadecyltrimethylammonium bromide turbidimetric method. Cellular uptake and subcellular localization of HA-liposomes were evaluated by flow cytometry and fluorescence microscopy. Mean particle sizes of HA-liposomes ranged from 120 to 180 nm and increased with the bigger size of HA. HA-liposome uptake correlated with HA MW (5-8 < 10-12 < 175-350 kDa), grafting density, and CD44 receptor density and exceeded that obtained with unconjugated plain liposomes. HA-liposomes were taken up into cells via lipid raft-mediated endocytosis, which is both energy- and cholesterol-dependent. Once within cells, HA-liposomes localized primarily to endosomes and lysosomes. The results demonstrate that cellular targeting efficiency of HA-liposomes depends strongly upon HA MW, grafting density, and cell surface receptor CD44 density. The results support a role of HA-liposomes for targeted drug delivery.

Synthesis of a Covalent Epirubicin-(C3-amide)-Anti-HER2/neuImmunochemotherapeutic Utilizing a UV-Photoactivated Anthracycline Intermediate

“…Second, maintaining maximum binding-avidity also directly facilitates efficient and more extensive internalization of epirubicin-(C 3 -amide)-[anti-HER2/neu] by mechanisms of ligand-induced receptor-mediated-endocytosis at HER2/neu overexpressed by mammary adenocarcinoma 7,26,30,120 and other neoplastic cell types. [121][122][123][124][125] Seemingly modest alterations in synthetic chemistry conditions, and elevations in chemotherapeutic molar-incorporation-indexes can profoundly influence the selective binding properties of immunoglobulin fractions. 29 Relatively higher anthracyclineimmunoglobulin molar-incorporation-indexes could have been attained during the synthesis of epirubicin-(C 3 -amide)-[anti-HER2/neu] utilizing succinimidyl 4,4-azipentanoate.…”

Effect of hyaluronic acid-binding to lipoplexes on intravitreal drug delivery for retinal gene therapy

“…Taken together we conclude that covalent HA coating 293 of lipoplexes outperforms electrostatic coating, both in terms of intravitreal mobility as its inherent 294 capacity to stimulate its own uptake and transfect ARPE19 cells. besides MW, has been thought to be grafting density (Qhattal and Liu, 2011), where it is reasoned that 316 the differences in CD44 affinity is due to the way free HA monomers are presented to the hyaladherins. 317…”

Peptide-mediated core/satellite/shell multifunctional nanovehicles for precise imaging of cathepsin B activity and dual-enzyme controlled drug release

“…The loading efficiency of HA on the FHMSNs@GNC was quantified using the published hexadecyltrimethylammonium bromide turbidimetric method. 28 The concentrations of protease enzyme Cat B in solution were detected by monitoring the fluorescence increase due to the cleavage of the Pep1 substrate by FHMSNs@GNC (unprotected nanovehicles) or FHMSNs@GNC@HA (self-protected nanovehicles).…”

“…Herein, HA, which specifically recognizes CD44 and RHAMM receptors overexpressed on the surface of some cancer cells, 27,28 was tethered to the nanovehicles, acting not only as a protecting shell but also as a target ligand. To verify the HA-mediated active targeting of the nanovehicles, HeLa cells (positive control) and NIH-3T3 cells (negative control) were incubated with different formulations containing Dox.…”

“…Then, hyaluronic acid (HA) molecules were assembled on the surface via peptide 2 (Pep2), which specifically binds to HA with a dissociation constant of 10 − 7 , 26 and HA then served as a shell that protected against nonspecific enzyme cleavage and an efficient target ligand for recognizing the CD44 and RHAMM receptors, which are overexpressed on the surface of cancer cells. 27,28 The above peptide-mediated core/satellite/shell nanocomplexes can be utilized as multifunctional nanovehicles for precise and dynamic imaging of Cat B and of controlled drug release, with the HA shell providing specific recognition and a protecting shell, the FHMSNs being drug carriers and signal generators, the GNC complexes being gatekeepers and signal inhibitors, and Cat B and hyaluronidase (HAase) 29 being dual openers and signal triggers. This self-protected peptide-based nanovehicle offers new avenues for the development of theranostic nanomedicine systems that could used for diagnosis, for therapy and for monitoring the delivery of drug molecules.…”