Engineering Multifunctional Cell Membrane-Derived Nanoparticles for Hepatic Steatosis and Hepatocellular Carcinoma Therapy

Zahid, Alap Ali

Engineering Multifunctional Cell Membrane-Derived Nanoparticles for Hepatic Steatosis and Hepatocellular Carcinoma Therapy

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Primary Zahid_Alap Ali_PhD_2026_thesis.pdf (7.53 MB)

Date

2026-03-11

Authors

Zahid, Alap Ali

Abstract

A significant challenge in pharmacotherapy is poor drug delivery, highlighting the need for platforms that efficiently deliver therapeutic agents to diseased tissues and improve efficacy. Biologically derived, engineered cell membrane-derived nanoparticles (CMNs) have emerged as promising drug-delivery systems in biomedical applications. Therefore, this thesis investigates the potential of CMNs as a drug-delivery platform to improve therapeutic efficacy for liver diseases, particularly metabolic dysfunction-associated steatotic liver disease (MASLD) and hepatocellular carcinoma (HCC). CMNs offer biocompatibility and tunable properties, with a favorable nanobiointerface that mimics native tissues, reduces immune recognition, and evades rapid clearance. Compared to synthetic nanoparticles or exosomes, CMNs are easier to produce, yield higher quantities, are noncytotoxic, and can be readily surface modified. In this thesis, CMNs were successfully fabricated via cell extrusion, with uniform shape and size confirmed by transmission electron microscopy (TEM) and other physicochemical characterization techniques. To assess CMN integrity via phosphatidylserine (PS) externalization (an ‘eat-me’ signal for phagocytic clearance), flow cytometry was performed. CMNs revealed only 1.13% PS in the outer leaflet compared with controls, indicating healthy “ghost” nanoparticles capable of prolonged systemic circulation. The CMNs demonstrated excellent drug-loading versatility and efficient cellular internalization capabilities, resulting in enhanced therapeutic efficacy. This was confirmed by loading a model drug (Rosuvastatin, RS), in which the RS-loaded CMN reduced hepatic steatosis in an in vitro hepatocyte steatosis model. Next, to achieve the targeted delivery of CMNs for hepatocyte-specific targeting, CMNs were prepared from red blood cells (RBCs) and functionalized with hepatocyte-specific targeting molecules to improve cellular specificity. The optimized CMNs were then loaded with the FDA-approved drug Resmetirom for MASLD therapy, significantly reducing hepatic steatosis in vitro. To further evaluate CMN's therapeutic potential, RBC-derived CMNs were applied to HCC treatment by incorporating the sonosensitizer indocyanine green (ICG) as a stimulus-responsive platform. In vitro evaluations showed that, upon ultrasound exposure, CMN-ICG produced 2.62-fold more reactive oxygen species (ROS) and induced mild hyperthermia (43-46 °C), ultimately leading to cancer cell death. Overall, these findings demonstrate the potential of CMNs as effective drug-delivery platforms and establish a foundation for future in vivo studies, supporting their clinical translation for liver diseases.

Summary for Lay Audience

A recent study reports that 90% of clinical drug development efforts yield suboptimal results, primarily because systemically administered drugs are unable to reach the site of injury effectively due to biological barriers and other physiological conditions present in the body. To overcome this challenge, nanoparticle-based drug-delivery platforms have been widely investigated, in which therapeutic agents are loaded into nanoparticle structures, enabling them to evade immune responses, prolong circulation time, and deliver the agents more effectively to the site of injury. Based on this promising strategy of nanoparticle-based drug-delivery platforms, we developed cell membrane-derived nanoparticles (CMNs) from mammalian cells. A mammalian cell is made up of phospholipid bilayers (called a cell membrane) that act as a cell wall. If this cell wall is fragmented into small pieces, it tends to self-assemble into a spherical shape. We leverage this self-assembly process to extrude cells through a series of filters, ultimately achieving CMNs with a size below 200 nm. The developed CMNs require no chemical interventions during preparation, exhibit promising cell-camouflaging properties (acting as ghost nanoparticles), and can target specific cells in the body. Given these benefits, this thesis focuses on developing CMNs from commercially available liver cells and murine red blood cells (RBCs) as a drug-delivery platform for the treatment of fatty liver disease and liver cancer. The thesis demonstrates that CMNs can load a variety of drugs into their lipid bilayer (cell membrane). CMNs are easier to produce and can be prepared from patient-specific cells for personalized therapy. The use of the designed CMNs is expected to: (Chapter 2) enhance drug therapeutic efficacy, (Chapter 3) specifically target liver cells, and (Chapter 4) non-invasive treatment of liver cancer using ultrasound-responsive CMNs that generate toxins to kill cancer cells. Overall, the findings from this thesis suggest that CMNs is a promising drug-delivery platform for enhancing therapeutic efficacy in fatty liver disease and can be used as an ultrasound-responsive nanoparticle system for treating liver cancer. We envision that this platform technology can be used to deliver various drugs for the treatment of other difficult-to-treat diseases.

Keywords

biomaterials, drug delivery, self-assembled nanoparticles, cell membrane nanoparticles, targeted nanoparticles, ultrasound-responsive nanoparticles, MASLD, liver cancer