An HK2 Antisense Oligonucleotide Induces Synthetic Lethality in HK1−HK2+ Multiple Myeloma

Abstract

A synthetically lethal precision medicine therapy for HK1-HK2+ multiple myeloma using an HK2 antisense oligonucleotide, metformin, and perhexiline. Most normal tissues Tolerated Tolerated HK1 + HK2 + multiple myeloma HK 2-A SO Me tfo rmi n Per hex ilin e HK1-HK2 + multiple myeloma Glucose-6-P Glucose-6-P Glucose Glucose HK1 HK2 HK2 Synthetically lethal Although the majority of adult tissues express only hexokinase 1 (HK1) for glycolysis, most cancers express hexokinase 2 (HK2) and many coexpress HK1 and HK2. In contrast to HK1 þ HK2 þ cancers, HK1 À HK2 þ cancer subsets are sensitive to cytostasis induced by HK2 shRNA knockdown and are also sensitive to synthetic lethality in response to the combination of HK2 shRNA knockdown, an oxidative phosphorylation (OXPHOS) inhibitor diphenyleneiodonium (DPI), and a fatty acid oxidation (FAO) inhibitor perhexiline (PER). The majority of human multiple myeloma cell lines are HK1 À HK2 þ. Here we describe an antisense oligonucleotide (ASO) directed against human HK2 (HK2-ASO1), which suppressed HK2 expression in human multiple myeloma cell cultures and human multiple myeloma mouse xenograft models. The HK2-ASO1/DPI/PER triple-combination achieved synthetic lethality in multiple myeloma cells in culture and prevented HK1 À HK2 þ multiple myeloma tumor xenograft progression. DPI was replaceable by the FDA-approved OXPHOS inhibitor metformin (MET), both for synthetic lethality in culture and for inhibition of tumor xenograft progression. In addition, we used an ASO targeting murine HK2 (mHK2-ASO1) to validate the safety of mHK2-ASO1/MET/PER combination therapy in mice bearing murine multiple myeloma tumors. HK2-ASO1 is the first agent that shows selective HK2 inhibition and therapeutic efficacy in cell culture and in animal models, supporting clinical development of this synthetically lethal combination as a therapy for HK1 À HK2 þ multiple myeloma. Significance: A first-in-class HK2 antisense oligonucleotide suppresses HK2 expression in cell culture and in in vivo, presenting an effective, tolerated combination therapy for preventing progression of HK1 À HK2 þ multiple myeloma tumors.

Hypoxia‐inducible hexokinase‐2 enhances anti‐apoptotic function via activating autophagy in multiple myeloma

“…The development of a specific inhibitor of HK2 may improve the prognosis of patients with refractory MM. Indeed, Xu et al 45 recently demonstrated the effectiveness of antisense oligos of HK2 against myeloma xenografted mice. However, because inhibition of the glycolytic system is also likely to damage normal tissues, it may be necessary to develop a drug delivery system that allows a HK2 inhibitor to exert its effect only in the bone marrow microenvironment.…”

Cyclin D1 targets hexokinase 2 to control aerobic glycolysis in myeloma cells

“…HK2 overexpression is observed in many solid tumors, and is associated with a poor prognosis 29 . Most MM tumor cells express HK2 30 . Our analysis of two cohorts of MM patients further indicated that high HK2 levels were correlated with a shorter EFS and OS.…”

“…Pharmacological agents that target glucose metabolism are generally highly cytotoxic 32 . However, the knockdown of HK2 expression with an antisense oligonucleotide (HK2-ASO) has a strong cytostatic effect on MM cell lines and in xenograft models 30 . Interestingly, HK2-ASO acts in synergy with an inhibitor of mitochondrial complex I (metformin), both in vitro and in vivo.…”

Enhanced antitumor effects of follicle-stimulating hormone receptor-mediated hexokinase-2 depletion on ovarian cancer mediated by a shift in glucose metabolism

“…Among HK family members, the expression of HK2 is relatively limited in normal adult tissues, including skeletal and adipose tissues, and HK2 is more catalytically efficient [ 9 ]. Moreover, HK2 is highly expressed in many tumor tissues and could be a potential target for antitumor therapy based on the promising results of preclinical studies [ 9 – 14 ].…”