Abstract 1998: Characterization of immunoliposomes for HER2-targeted delivery of the dual Rac/Cdc42 inhibitor MBQ-167
Authors: Velazquez Luis E. , Dharmawardhane Suranganie 
Published: April 2023

Breast cancer is the second leading cause of mortality among women in the US. Among the various subtypes, human epidermal growth factor receptor 2 (HER2) positive breast cancer shows a very aggressive and invasive phenotype. This subtype is characterized by the overexpression of HER2 receptor on the cell surface that upon ligand binding and dimerization, leads to overactive cell signaling that promotes cell proliferation and metastasis. Even though therapies have been developed to treat this type of breast cancer by targeting HER2 and preventing dimerization (eg. Trastuzumab), patients can present with acquired or intrinsic therapy resistance. One of the mechanisms of resistance is the compensation of intracellular signaling from other receptors. This signaling converges on guanine nucleotide exchange factors (GEFs) that activate Rac and Cdc42 by exchanging GDP for GTP, thus activating downstream effectors that modulate the actin cytoskeleton to promote cell migration and invasion. Therefore, targeting Rac and Cdc42 activation selectively in HER2 positive breast cancer is a promising strategy for overcoming HER2-targeted therapy resistance. Previously, we characterized the dual Rac/Cdc42 inhibitor MBQ-167 that inhibits Rac and Cdc42 activity with IC50s of 103nM and 78nM, respectively. However, there is a need to develop selective delivery systems that transport MBQ-167 directly into HER2-positive breast cancer cells. Our objective is to deliver MBQ-167 selectively into HER2-positive cells using liposomes coated with Trastuzumab, a clinically used monoclonal antibody that targets HER2. We conjugated Trastuzumab to a lipid linker (DSPE-PEG-Maleimide) by reacting Trastuzumab with 2-iminothiolane (Traut’s reagent) under nitrogenated (low oxygen) conditions and then mixing with the lipid linker overnight. This reaction was characterized by measuring the thiol groups formed after the reaction with Traut’s reagent and after mixing with the lipid, followed by mixing the Trastuzumab-lipid conjugate with liposomes containing MBQ-167. To quantify the amount of MBQ-167 in the liposomes, we determined the excitation/emission parameters of the molecule and measured the concentration of MBQ-167 by fluorescence. We found an increase in thiol groups after the reaction with Traut’s reagent, which decreased after mixing with the lipids, suggesting the formation of the DSPE-PEG-Trastuzumab conjugate. Additionally, we determined the excitation/emission parameters (320nm/430nm), quantified a lower limit of detection (LLOD) at 0.1mM, and calculated an encapsulation efficiency of 97% (530μM). Future studies include testing our formulation in vitro and in vivo in HER2+ breast cancer models.

Citation Format: Luis E. Velazquez, Suranganie Dharmawardhane. Characterization of immunoliposomes for HER2-targeted delivery of the dual Rac/Cdc42 inhibitor MBQ-167 [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 1998.