Reassembly of 89 Zr-Labeled Cancer Cell Membranes into Multicompartment Membrane-Derived Liposomes for PET-Trackable Tumor-Targeted Theranostics

Abstract

Nanoengineering of cell membranes holds great potential to revolutionize tumor-targeted theranostics, owing to their innate biocompatibility and ability to escape from the immune and reticuloendothelial systems. However, tailoring and integrating cell membranes with drug and imaging agents into one versatile nanoparticle are still challenging. Here, multicompartment membrane-derived liposomes (MCLs) are developed by reassembling cancer cell membranes with Tween-80, and are used to conjugate ⁸⁹ Zr via deferoxamine chelator and load tetrakis(4-carboxyphenyl) porphyrin for in vivo noninvasive quantitative tracing by positron emission tomography imaging and photodynamic therapy (PDT), respectively. Radiolabeled constructs, ⁸⁹ Zr-Df-MCLs, demonstrate excellent radiochemical stability in vivo, target 4T1 tumors by the enhanced permeability and retention effect, and are retained long-term for efficient and effective PDT while clearing gradually from the reticuloendothelial system via hepatobiliary excretion. Toxicity evaluation confirms that the MCLs do not impose acute or chronic toxicity in intravenously injected mice. Additionally, ⁸⁹ Zr-labeled MCLs can execute rapid and highly sensitive lymph node mapping, even for deep-seated sentinel lymph nodes. The as-developed cell membrane reassembling route to MCLs could be extended to other cell types, providing a versatile platform for disease theranostics by facilely and efficiently integrating various multifunctional agents.

Keywords: cancer cell membranes; cancer theranostics; membrane-derived liposomes; positron emission tomography; targeted drug delivery.

Figure 1

Preparation and ⁸⁹Zr-labeling of MCLs. (A) A schematic illustration of MCL fabrication. (B) The particle diameter of tween-80 micelles, cell membrane nano-vesicle (CMVs), and MCLs. (C, D) TEM imaging of CMVs and MCLs. (E) Time-dependent ⁸⁹Zr labeling yields of Df-MCLs. The inset photo represents PET images of different fractions collected after PD-10 purification of Df-MCLs incubated with ⁸⁹Zr for 2 h. (F) Autoradiograph of TLC plates of ⁸⁹Zr-Df-MCLs at different incubation time points. (G) Serum stability study of ⁸⁹Zr-Df-MCLs.

Figure 2

Misdistribution and hepatobiliary excretion of ⁸⁹Zr-Df-MCLs. (A) In vivo serial PET images of mice taken at various time points (0.5, 1, 2, 5, 12, 24, 48, and 72 h) post intravenous injection of ⁸⁹Zr-Df-MCLs. Organ uptake was presented as %ID/g. (B) The hepatic clearance of ⁸⁹Zr-Df-MCLs excreted in the feces at various time points p.i. The data is presented as %ID. (n=3) (C) PET images of feces taken at various time points. Ex vivo PET imaging of main organs at different time points p.i. (D) 1 h, and (E) 24 h.

Figure 3

Tumor targeting and in vivo anti-tumor therapy of ⁸⁹Zr-Df-MCLs. (A) In vivo PET images of 4T1 tumor-bearing mice taken at various time points post intravenous injection of ⁸⁹Zr-Df-MCLs. (B) Quantification of ⁸⁹Zr-Df-MCLs uptake in the tumor at various time points p.i. The unit is the percentage of injected dose per gram of tissue (%ID/g) (n=3). (C) Misdistribution of ⁸⁹Zr-Df-MCLs 72 h after intravenous injection into 4T1 tumor-bearing mice as determined by ⁸⁹Zr radioactivity measurements in various organs (n=3). (D) Body weight measurements after various treatments showed no significant toxicity (n=5–6). (E) Tumor growth profiles of 4T1 tumors after each treatment. For the combination treatment group, six mice injected with TCPP-MCLs were irradiated with 660 nm laser (50 mW/cm², 40 min) at 24 h p.i.. Two groups of mice were used as controls: untreated (PBS) (n=5); and TCPP-MCLs only without laser treatment (n=6). (***p < 0.001).

Figure 4

Lymph node PET imaging of ⁸⁹Zr-Df-MCLs. (A) In vivo lymph node imaging with PET upon footpad injection of ⁸⁹Zr-Df-MCLs at different time points. (B) Quantification of ⁸⁹Zr-Df-MCLs uptake by the mouse footpad and lymph nodes at various time points. (C) In vivo and Ex vivo quantification of ⁸⁹Zr-Df-MCLs uptake by the mouse footpad and lymph nodes at 24 h time point after footpad injection. (D) Ex vivo PET imaging of lymph nodes and mouse at 24 h p.i.