Targeted Liposomal Chemotherapies to Treat Triple-Negative Breast Cancer

Abstract

Triple-negative breast cancers (TNBCs) are highly aggressive and recurrent. Standard cytotoxic chemotherapies are currently the main treatment options, but their clinical efficacies are limited and patients usually suffer from severe side effects. The goal of this study was to develop and evaluate targeted liposomes-delivered combined chemotherapies to treat TNBCs. Specifically, the IC50 values of the microtubule polymerization inhibitor mertansine (DM1), mitotic spindle assembly defecting taxane (paclitaxel, PTX), DNA synthesis inhibitor gemcitabine (GC), and DNA damage inducer doxorubicin (AC) were tested in both TNBC MDA-MB-231 and MDA-MB-468 cells. Then we constructed the anti-epidermal growth factor receptor (EGFR) monoclonal antibody (mAb) tagged liposomes and confirmed its TNBC cell surface binding using flow cytometry, internalization with confocal laser scanning microscopy, and TNBC xenograft targeting in NSG female mice using In Vivo Imaging System. The safe dosage of anti-EGFR liposomal chemotherapies, i.e., <20% body weight change, was identified. Finally, the in vivo anti-tumor efficacy studies in TNBC cell line-derived xenograft and patient-derived xenograft models revealed that the targeted delivery of chemotherapies (mertansine and gemcitabine) can effectively inhibit tumor growth. This study demonstrated that the targeted liposomes enable the new formulations of combined therapies that improve anti-TNBC efficacy.

Targeted EV to Deliver Chemotherapy to Treat Triple-Negative Breast Cancers

“…The relevant literature, in addition to [36,37,43,44,57] this study, revealed high expression of the surface receptors EGFR and CD47 in TNBC patient tissues. This study constructed an EGFR/CD47 dual-targeted mAb-EV to deliver the natural compound Ver-A for TNBC treatment.…”

“…The TNBC PDX xenograft donor mice were purchased from The Jackson Laboratory and maintained at low passages of 2-4 in host NSG mice following our established protocol [57]. Briefly, either fresh harvested tumor or fresh frozen tumor tissues were minced into small fragments (<1 mm 3 ) and subcutaneously (s.c.) injected into the right flank of NSG mouse.…”

Process Improvement of Adeno-Associated Virus Production

“…The transfected HKE 293AAV cells were harvested and centrifuged at 48-60 h for further AAV clarification. 3) Cationic liposomes transfection: We synthesized cationic liposomes following similar synthetic procedure of neutral liposomes (Si et al, 2021) with modifications: mixing 7 µmole 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE), 7 µmole 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP) and 10 ml of chloroform and evaporating at 60 °C and 50 rpm for 1 h. The generated liposomes were analyzed using NanoSight, which showed concentration of 3.2 × 10 11 particles/ml and mean size of 92.6 ± 0.4 nm. Total of 30 µg three plasmids (10 µg each plasmid) were mixed with 900 µL of cationic liposomes, incubated on ice for 30 min, and mixed with Opti-MEM medium with volume ratio of 1:1.…”

Targeted Extracellular Vesicles Delivered Verrucarin A to Treat Glioblastoma

“…The brain GBM tissue arrays, including 35 cases and 70 cores, were purchased from US Biomax (Derwood, MD, USA). IHC staining was performed to identify the surface receptor in patient tissues as we described before [ 40 , 48 ]. The TMA slides were stained with rabbit anti-human EGFR mAb (Abcam, Waltham, MA, USA) and counterstained with hematoxylin.…”

“…The sections of major organs harvested from the in vivo treatment with mAb-EV-Ver-A were stained with H&E. The detailed staining procedure has been reported in our previous publications [ 40 , 48 ].…”