Protein Corona Fingerprints of Liposomes: New Opportunities for Targeted Drug Delivery and Early Detection in Pancreatic Cancer

Abstract

Pancreatic ductal adenocarcinoma (PDAC) is the fourth cause of cancer-related mortality in the Western world and is envisaged to become the second cause by 2030. Although our knowledge about the molecular biology of PDAC is continuously increasing, this progress has not been translated into better patients’ outcome. Liposomes have been used to circumvent concerns associated with the low efficiency of anticancer drugs such as severe side effects and damage of healthy tissues, but they have not resulted in improved efficacy as yet. Recently, the concept is emerging that the limited success of liposomal drugs in clinical practice is due to our poor knowledge of the nano–bio interactions experienced by liposomes in vivo. After systemic administration, lipid vesicles are covered by plasma proteins forming a biomolecular coating, referred to as the protein corona (PC). Recent studies have clarified that just a minor fraction of the hundreds of bound plasma proteins, referred to as “PC fingerprints” (PCFs), enhance liposome association with cancer cells, triggering efficient particle internalization. In this study, we synthesized a library of 10 liposomal formulations with systematic changes in lipid composition and exposed them to human plasma (HP). Size, zeta-potential, and corona composition of the resulting liposome–protein complexes were thoroughly characterized by dynamic light scattering (DLS), micro-electrophoresis, and nano-liquid chromatography tandem mass spectrometry (nano-LC MS/MS). According to the recent literature, enrichment in PCFs was used to predict the targeting ability of synthesized liposomal formulations. Here we show that the predicted targeting capability of liposome–protein complexes clearly correlate with cellular uptake in pancreatic adenocarcinoma (PANC-1) and insulinoma (INS-1) cells as quantified by flow-assisted cell sorting (FACS). Of note, cellular uptake of the liposomal formulation with the highest abundance of PCFs was much larger than that of Onivyde®, an Irinotecan liposomal drug approved by the Food and Drug Administration in 2015 for the treatment of metastatic PDAC. Given the urgent need of efficient nanocarriers for the treatment of PDAC, we envision that our results will pave the way for the development of more efficient PC-based targeted nanomaterials. Here we also show that some BCs are enriched with plasma proteins that are associated with the onset and progression of PDAC (e.g., sex hormone-binding globulin, Ficolin-3, plasma protease C1 inhibitor, etc.). This could open the intriguing possibility to identify novel biomarkers.

CXCL5 Modified Nanoparticle Surface Improves CXCR2+ Cell Selective Internalization

“…The search for nanomaterials (NMs) with the ability to travel to specific biological tissues is constantly increasing. Several nanoparticles (NP) engineering methods [1] as well as specific corona formation [2] have been tested thus far. However, many physical, chemical, or biological barriers limit their successful application.…”

Mastering the Tools: Natural versus Artificial Vesicles in Nanomedicine

“…[102] Through this method, Palchetti et al, in 2019, developed a protocol to finely control the composition, rigidity, and dimension of ten different liposomes formulations, allowing the so-produced liposomes to safely interact with plasma proteins derived from human serum. [103] In terms of their structure, liposomes can be classified as multilamellar vesicles (MLVs) if they are composed of several lipid bilayers (their size is in the range of 50-1000 nm), or unilamellar vesicles (ULVs) if they are formed by a single lipid bilayer. ULVs have permitted the development of many commercialized drug delivery products.…”

“…Indeed, given their higher flexibility in production and easier functionalization, polymersomes can be envisaged as an advantageous and cheaper alternative to liposomes. Extracellular vesicles + Difficult massive production and purification; biomolecules as only building blocks; limited tunability of the physicochemical properties [ 87,240] +++ Generally safe; extraction from biological sources with high biocompatibility [80,83] ++ Excellent loading of biomolecules; challenging loading of synthetic molecules; good delivery efficiency;rapid clearance [ 186,187] Liposomes ++ Easy production and purification; pre-and postsynthetic functionalization; tunable physicochemical properties [ 124,161] +++ Generally safe, unless modified with charged phospholipids; similar composition to EVs, but without cell-mediated modifications [103,213] ++ Good natural and synthetic molecules loading; preassembly and postassembly loading; good delivery efficiency; tunable clearance properties [ 198,199] Polymersomes +++ Customizable massive production; pre-and postsynthetic functionalization; tunable physicochemical properties [ 136,155] + Possible accumulations in the body; possibility to introduce biodegradable polymers [229,230] + Preassembly and postassembly loading; cell uptake dependent on the vesicle and cell membrane characteristic; tunable clearance properties [ 222,224]…”

Effect of Protein Corona on The Transfection Efficiency of Lipid-Coated Graphene Oxide-Based Cell Transfection Reagents

“…After the washes, the pellets were suspended in 20 µL of Laemmli loading buffer 1x, boiled for 10 min at 100 • C and centrifuged at 18,620 RCF for 15 min at 4 • C. Supernatants were collected and loaded on a stain-free gradient polyacrylamide gel (4-20% TGX precast gels, Bio-Rad, Hercules, CA, USA) and run at 100 V for about 150 min. Finally, gel images were acquired with a ChemiDoc™ gel imaging system (Bio-Rad, CA, USA) and processed by means of custom MatLab scripts (MathWorks, Natick, MA, USA) [26].…”