Arginine Deiminase Induces Immunogenic Cell Death and Is Enhanced by N-acetylcysteine in Murine MC38 Colorectal Cancer Cells and MDA-MB-231 Human Breast Cancer Cells In Vitro

Abstract

The use of arginine deiminase (ADI) for arginine depletion therapy is an attractive anticancer approach. Combination strategies are needed to overcome the resistance of severe types of cancer cells to this monotherapy. In the current study, we report, for the first time, that the antioxidant N-acetylcysteine (NAC), which has been used in therapeutic practices for several decades, is a potent enhancer for targeted therapy that utilizes arginine deiminase. We demonstrated that pegylated arginine deiminase (ADI-PEG 20) induces apoptosis and G0/G1 phase arrest in murine MC38 colorectal cancer cells; ADI-PEG 20 induces Ca2+ overload and decreases the mitochondrial membrane potential in MC38 cells. ADI-PEG 20 induced the most important immunogenic cell death (ICD)-associated feature: cell surface exposure of calreticulin (CRT). The antioxidant NAC enhanced the antitumor activity of ADI-PEG 20 and strengthened its ICD-associated features including the secretion of high mobility group box 1 (HMGB1) and adenosine triphosphate (ATP). In addition, these regimens resulted in phagocytosis of treated MC38 cancer cells by bone marrow-derived dendritic cells (BMDCs). In conclusion, we describe, for the first time, that NAC in combination with ADI-PEG 20 not only possesses unique cytotoxic anticancer properties but also triggers the hallmarks of immunogenic cell death. Hence, ADI-PEG 20 in combination with NAC may represent a promising approach to treat ADI-sensitive tumors while preventing relapse and metastasis.

Targeting the Integrated Stress Response in Cancer Therapy

“…These elF2α kinase activators have been studied in cancer therapy. For example, Halofuginone and arginine deiminase were found to inhibit tumor growth, development and metastasis either as single agents or in combination with 5-FU or radiation (Abramovitch et al, 2004;Kim et al, 2009;Cook et al, 2010;Spector et al, 2010;Lamora et al, 2015;Brin et al, 2018;Singh et al, 2019;Wang et al, 2020;Huang and Hu, 2021). Our laboratory has identified two small molecules PG3-Oc (Tian et al, 2021) and ONC201 (Kline et al, 2016;Ishizawa et al, 2016) that suppress tumor growth through increased ISR signaling.…”

Arginine Signaling and Cancer Metabolism

“…Furthermore, using murine MC38 colon cell model, it was shown that ADI-PEG20 is able to induce apoptotic and immunogenic cell death with the phenotypes of cell exposure of calreticulin, the release of HMGA1 and ATP, which is enhanced by N-acetylcystein [ 106 ]. These “eat me” signals resulted in increased phagocytosis by bone marrow-derived dendritic cells.…”

Targeting HMGB1: An available Therapeutic Strategy for Breast Cancer Therapy

“…In addition, Somma et al found that G-quadruplex binders induced ICD in MDA-MB-231 breast cancer cells, accompanied by T-cell activation 140 . Furthermore, Huang et al indicated that the antioxidant N-acetylcysteine (NAC) enhanced not only the antitumor activity of polyglycolylated arginine deiminase (ADI-PEG20), but also the features of ICD, including HMGB1 and ATP secretion 19 . Nevertheless, the ICD inducers in clinical trials for breast cancer are limited so far.…”