Systematic identification of arsenic-binding proteins reveals that hexokinase-2 is inhibited by arsenic

Abstract

Arsenic is highly effective for treating acute promyelocytic leukemia (APL) and has shown significant promise against many other tumors. However, although its mechanistic effects in APL are established, its broader anticancer mode of action is not understood. In this study, using a human proteome microarray, we identified 360 proteins that specifically bind arsenic. Among the most highly enriched proteins in this set are those in the glycolysis pathway, including the rate-limiting enzyme in glycolysis, hexokinase-1. Detailed biochemical and metabolomics analyses of the highly homologous hexokinase-2 (HK2), which is overexpressed in many cancers, revealed significant inhibition by arsenic. Furthermore, overexpression of HK2 rescued cells from arsenic-induced apoptosis. Our results thus strongly implicate glycolysis, and HK2 in particular, as a key target of arsenic. Moreover, the arsenic-binding proteins identified in this work are expected to serve as a valuable resource for the development of synergistic antitumor therapeutic strategies.arsenic trioxide | human proteome microarray | glycolysis | hexokinase-2IA rsenic and its derivatives have been applied as therapy for a variety of diseases for more than 2,200 y (1). To date, the disease most successfully treated with these types of compounds is acute promyelocytic leukemia (APL). Administration of arsenic trioxide (ATO) combined with all-trans retinoic acid has demonstrated a remarkable 5-y overall survival rate of 85-90% as a consequence of the dramatic down-regulation of the key protein driving APL tumorigenicity, promyelocytic leukemia-retinoic acid receptor α (PML-RARα) (2). However, ATO also has been found to be effective against many other hematologic malignancies and solid tumors. For example, together with imatinib it is a promising treatment for chronic myelocytic leukemia (3), and it has been used alone with some success to treat multiple myeloma (4), myelodysplasia syndrome (5), and non-Hodgkin lymphoma (6). ATO also is under clinical investigation as a possible medication for lung cancer, hepatocellular carcinoma, melanoma, renal cell carcinoma, and colorectal cancer (https://www.clinicaltrials.gov/). At the cellular level, ATO has been shown to inhibit significantly the growth of almost all the cell lines (59 of 60) in the US National Cancer Institute anticancer drug screen that spans nine different tumor types (7). Thus ATO is one of the most promising broadly effective medications against cancer.Although its mode of action in APL is well established, the underlying mechanisms by which ATO acts in other types of cancers remain poorly understood. A variety of systematic studies, including studies that provided transcriptomic, chemogenomic, or proteomic characterizations (8, 9), have been performed to gain a better understanding of this broader anticancer activity of ATO. However, these studies examined only the cellular consequences of treatment with ATO without identifying the primary proteins directly bound and modulated by ATO. Knowledge of ATO...

Construction of a metabolomics profile of arsenic trioxide effect in gastric carcinoma cell line SGC7901

“…But, as demonstrated by a variety of other studies, cell lines of solid tumors were usually treated with 1-2 μM ATO for 12-24 h, and significant growth inhibition and apoptosis were observed in many of these cell lines, such as MGC803 [39], MCF-7, HeLa, and HIC [40]. According to our recent study [38], significant growth inhibition was also observed in SGC7901 cells after 12-24 h of ATO treatment at a dose of 2 μM. Our study employing the human protein microarray showed that ATO-binding proteins are involved in a variety of cellular signaling pathway, such as apoptosis, protein kinase, and acetylation/deacetylation pathway.…”

“…As such, we could then correlate the metabolomics data with transcriptomics and proteomics. This multiple level investigation would definitely help us to understand MOA of ATO at a more systematical and comprehensive level [38].…”

“…Several studies have shown that ATO perturbation in solid tumor may have different mechanisms from that of hematological carcinoma [34][35][36][37]. According to our recent study [38], ATO binds to >300 proteins. Usually ATO causes activity loss of the proteins that it binds with, but it will not cause direct change of the targeting proteins at either mRNA level or protein level.…”

“…Our study employing the human protein microarray showed that ATO-binding proteins are involved in a variety of cellular signaling pathway, such as apoptosis, protein kinase, and acetylation/deacetylation pathway. Interestingly, ATO-binding proteins were highly enriched in glycolysis pathway, particularly, overexpression of tumor-specific glycolysis rate-limiting enzyme hexokinase-2 could dramatically rescue ATO-treated cells from apoptosis [38].…”

miR-194 as predictive biomarker of responsiveness to neoadjuvant chemoradiotherapy in patients with locally advanced rectal adenocarcinoma

“…In addition, analysing the CTD results, the identified compounds have to be subdivided between toxic and therapeutic because the database structure does not provide for a subdivision a priori. On this basis, one of the most interesting compounds was 4-aminophenylarsenoxide, an arsenic derivative, already used as a chemotherapeutic together with imatinib in the treatment of chronic myeloid leukaemia, that has been used alone with some success to treat multiple myeloma, myelodysplasia syndrome and non-Hodgkin’s lymphoma 33. Another intriguing compound was PD 0325901, a second-generation MEK inhibitor, currently in phase I trial for treating patients with solid tumours that have spread to other parts of the body or cannot be removed by surgery (National Cancer Institute, https://www.cancer.gov).…”