The Circadian Clock Regulates Metabolic Phenotype Rewiring Via HKDC1 and Modulates Tumor Progression and Drug Response in Colorectal Cancer

Abstract

An endogenous molecular clockwork drives various cellular pathways including metabolism and the cell cycle. Its dysregulation is able to prompt pathological phenotypes including cancer. Besides dramatic metabolic alterations, cancer cells display severe changes in the clock phenotype with likely consequences in tumor progression and treatment response. In this study, we use a comprehensive systems-driven approach to investigate the effect of clock disruption on metabolic pathways and its impact on drug response in a cellular model of colon cancer progression. We identified distinctive time-related transcriptomic and metabolic features of a primary tumor and its metastatic counterpart. A mapping of the expression data to a comprehensive genome-scale reconstruction of human metabolism allowed for the in-depth functional characterization of 24 h-oscillating transcripts and pointed to a clock-driven metabolic reprogramming in tumorigenesis. In particular, we identified a set of five clock–regulated glycolysis genes, ALDH3A2, ALDOC, HKDC1, PCK2, and PDHB with differential temporal expression patterns. These findings were validated in organoids and in primary fibroblasts isolated from normal colon and colon adenocarcinoma from the same patient. We further identified a reciprocal connection of HKDC1 to the clock in the primary tumor, which is lost in the metastatic cells. Interestingly, a disruption of the core-clock gene BMAL1 impacts on HKDC1 and leads to a time-dependent rewiring of metabolism, namely an increase in glycolytic activity, as well as changes in treatment response. This work provides novel evidence regarding the complex interplay between the circadian clock and metabolic alterations in carcinogenesis and identifies new connections between both systems with pivotal roles in cancer progression and response to therapy.

The importance of determining circadian parameters in pharmacological studies

“…Circadian studies have been performed in a multitude of in vitro cell models, ranging from rodent and human immortalized cell lines,such as NIH/3T3‐immortalized mouse embryo fibroblasts (Akashi & Nishida, ) and rat‐1 fibroblasts (Balsalobre, Damiola, & Schibler, ; Izumo, Sato, Straume, & Johnson, ), or human cancer lines, as U2OS osteosarcoma cells (Relógio et al, ); primary cultures, as mouse neonatal cardiomyocytes (du Pré et al, ) and human fibroblasts (Brown et al, ), blood monocytes (Keller et al, ), hair follicle cells (Akashi et al, ), or keratinocytes (Sandu et al, ); to organoids (Fuhr et al, ) and tissue explants, as mouse fetal adrenal gland (Ungar & Halberg, ) or SCN explants (Hughes et al, ).…”

“…Technological advances have allowed longitudinal measurements in real‐time living cells with a higher temporal resolution, allowing assessment of the persistence and dynamics of molecular rhythms. Automated luminometer devices with photomultiplier tubes, as LumiCycle, are commonly used for real‐time recording (Fuhr et al, ). These systems employ photomultiplier tubes as light detectors, providing extremely high sensitivity and low‐noise data, particularly suitable for data acquisition of extremely dim luc‐based bioluminescence (Ramanathan et al, ).…”

“…Also, metabolic alterations resulting from a dysregulated clock were detected with a computational/experimental high‐throughput approach using a human colon cancer model. The authors identified well‐known metabolic genes to be under circadian control and further expanded their findings to the analysis of metabolic alterations due to the action of different drugs in a time‐dependent manner (Fuhr et al, ). Another large‐scale study, which characterized the variation in expression of clock genes across 32 cancer types, demonstrated that alterations of core‐clock genes are associated with key oncogenic pathways and tumour stage and correlate with patient survival (Ye et al, ).…”

“…These TFs, via transcriptional and translational feedback loops, regulate their own transcription, which leads to ≈24‐hr oscillations in their gene and protein expression in all cells, tissues, and organs. In addition, the CCN elements coordinate the transcription of many other genes, clock‐controlled genes (CCGs), in a circadian‐ and organ‐specific manner, regulating a variety of biological processes, at both the cell and tissue levels, such as cell division, bioenergetics, autophagy, metabolism, redox balance, hormone regulation, and immune responses (El‐Athman et al, ; Fuhr et al, ; Relógio et al, ; Scrima et al, ; Zhang, Lahens, Ballance, Hughes, & Hogenesch, ; reviewed in Pilorz et al, ). These cellular peripheral clocks, distributed throughout the body, are further coordinated by a central pacemaker, located at the suprachiasmatic nuclei (SCNs) in the hypothalamus (Albrecht, ).…”

The tumour suppressing role of the circadian clock

“…Fuhr et al recently found that 16.0% (3,998) of all genes were oscillating in SW480 colon cancer cells and 14.8% (3,693) were oscillating in its metastatic counterpart, SW620. However, only 5.5% (1,385) of all genes oscillate in both cell lines, which point to a switch in the 24 h oscillating gene expression profile as cells progress from primary tumour to metastatic .…”