The metal complex copper diethyldithiocarbamate (CuET) induces cancer cell death by inhibiting protein degradation and induces proteotoxic stress, making CuET a promising cancer therapeutic. However, no clinical formulation of CuET exists to date as the drug is insoluble in water and exhibits poor bioavailability. To develop a scalable formulation, nanoliposomal (LP) CuET was synthesized using ethanol injection as a facile one-step method that is suitable for large-scale manufacturing. The nanoparticles are monodispersed, colloidally stable, and approximately 100 nm in diameter with an encapsulation efficiency of over 80%. LP-CuET demonstrates excellent stability in plasma, minimal size change, and little drug release after six-month storage at various temperatures. Additionally, melanoma cell lines exhibit significant sensitivity to LP-CuET and cellular uptake occurs predominantly through endocytosis in YUMM 1.7 cancer cells. Intracellular drug delivery is mediated by vesicle acidification with more nanoparticles being internalized by melanoma cells compared with RAW 264.7 macrophages. Additionally, the nanoparticles preferentially accumulate in YUMM 1.7 tumors where they induce cancer cell death in vivo. The development and characterization of a stable and scalable CuET formulation illustrated in this study fulfils the requirements needed for a potent clinical grade formulation.

In this paper, we introduce a SAW-based platform that generates acoustic microstreaming and Bjerknes forces to enhance the delivery of nanoparticles on the spheroid's surface and improve their penetration into deeper layers of spheroids.

Introduction: Colorectal cancer (CRC) is the second leading cause of cancer-related death worldwide, contributing to one million deaths yearly. While survival is expected in early-stage disease, the 5-year survival rate is only 15% in metastatic CRC (mCRC). Frequently patients develop resistance to their chemotherapy regimens resulting in CRC metastatic lesions to appear in the lungs, liver, and brain. In recent years, the development of immunotherapy has become a promising avenue for the treatment of cold-tumor environments such as CRC. Indeed, dense infiltration of lymphocytes in CRC correlates with longer patient survival. Thus, patients suffering from mCRC would benefit from the combination of classical chemotherapy/radiotherapy with small immunomodulatory molecules capable of overcoming cancer cell resistance to therapy through chemo- and radio-sensitization. Recently, Antabuse’s metabolite dithiocarb-copper complex (CuET) was found to have anti-cancer activity. Here, we report for the first time on the immunomodulatory properties of CuET in the context of T/NK cell antitumor response in mCRC. Methods: To assess CuET anticancer activity in vitro, we performed survival curve, colony-formation, and migration-invasion assays in the murine colorectal cancer cell lines MC-38 and CT-26, and in the human KRAS mutant HCT116 cell line. We assessed in vivo efficacy of CuET in ectopic and metastatic MC-38 and CT-26 models in respective C57BL/6 and BALB/c mice. We characterized tumor growth, mice survival, and tumor immunohistochemistry. We evaluated CuET’s immunomodulatory properties through immunophenotyping of mouse and human T/NK cell receptors as well as corresponding tumor cell ligands after CuET treatment. We assessed CuET’s effect on the functional anticancer cytotoxicity of human PBMC-derived T/NK cells. Results: In vitro, CuET significantly inhibits viability, clonogenicity, and migration of tumor cells MC-38, CT-26, and HCT116 with IC50 values in the nanomolar range. In vivo, systemic treatment with CuET significantly reduces tumor growth in mice and prolongs survival (p=0.0041) when compared to controls. CuET significantly inhibits MC-38 liver metastasis growth in C57BL/6 mice. CuET recruits macrophages and T lymphocytes inside tumor cores and induces apoptosis. CuET induces the expression of NKG2D activating receptors in NK and T lymphocytes, as well as the expression of the corresponding ligands MICA/B and ULBP1/2 on tumor cells, enhancing the T/NK cell cytotoxic response anticancer. Conclusion: CuET enhances the functional antitumor cytotoxic activity of NK and T lymphocytes in mice and in human PBMC. In summary, our findings demonstrate that Antabuse-derived copper-diethyldithiocarbamate, CuET, is a potent immunomodulator which could benefit patients with mCRC. Citation Format: Daciana Catalina Dumut, Ivo Frydrych, Miroslav Popper, Dusan Garic, Radu Alexandru Paun, Amanda Centorame, Olivia Canavan, Juhi Shah, Martin Mistrik, Petr Dzubak, Jiri Bartek, Marian Hajduch, Juan Bautista DeSanctis, Danuta Radzioch. Disulfiram metabolite modulates NK and T cell cytotoxicity against metastatic colorectal cancer through tumor derived NKG2D ligands [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 2329.

Cystic fibrosis (CF) is the most common autosomal recessive genetic disease in Caucasians, affecting more than 100,000 individuals worldwide. It is caused by pathogenic variants in the gene encoding CFTR, an anion channel at the plasma membrane of epithelial and other cells. Many CF pathogenic variants disrupt the biosynthesis and trafficking of CFTR or reduce its ion channel function. The most frequent mutation, loss of a phenylalanine at position 508 (F508del), leads to misfolding, retention in the endoplasmic reticulum, and premature degradation of the protein. The therapeutics available for treating CF lung disease include antibiotics, mucolytics, bronchodilators, physiotherapy, and most recently CFTR modulators. To date, no cure for this life shortening disease has been found. Treatment with the Triple combination drug therapy, TRIKAFTA®, is composed of three drugs: Elexacaftor (VX-445), Tezacaftor (VX-661) and Ivacaftor (VX-770). This therapy, benefits persons with CF, improving their weight, lung function, energy levels (as defined by reduced fatigue), and overall quality of life. We examined the effect of combining LAU-7b oral treatment and Triple therapy combination on lung function in a F508deltm1EUR mouse model that displays lung abnormalities relevant to human CF. We assessed lung function, lung histopathology, protein oxidation, lipid oxidation, and fatty acid and lipid profiles in F508deltm1EUR mice.