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This work describes a comprehensive systems biology approach to couple circadian molecular processes with tumourigenesis., Diese Arbeit beschreibt einen systembiologischen Ansatz zur Kopplung zirkadianisch-molekularer Prozesse mit Tumorgenese.

Die zirkadiane Uhr ist ein endogenes Zeitmesssystem, das die Anpassung physiologischer Prozesse an die geophysikalische Zeit ermöglicht. Die zirkadiane Uhr besteht aus einem zentralen Schrittmacher und peripheren Uhren in jeder Zelle. Bei Säugern ist eine bestimmte Anzahl von Uhr-Genen in regulatorischen Schleifen miteinander verbunden, wodurch Oszillationen in der Expression der Uhr-Gene sowie in zahlreichen Zielgenen erzeugt werden. Zielgene der zirkadianen Uhr sind unter anderem an zellulären Prozessen beteiligt, die eine Rolle bei der Tumorentstehung und -progression spielen. Funktionsstörungen der zirkadianen Uhr stehen im Zusammenhang mit verschiedenen Krankheitsbildern, unter anderem Krebs. Ziel dieses Projekts war es, die Rolle der zirkadianen Uhr bei der Tumorentstehung und entwicklung zu untersuchen. Der Fokus lag dabei auf tumorspezifischen Stoffwechselwegen. Die Rolle einer deregulierten zirkadianen Uhr wurde in einem in vitro Zellmodel untersucht. Die SW480 Zelllinie wurde aus einem Primärtumor isoliert und die SW620 Zelllinie aus einer Lymphknotenmetastase desselben Patienten. Die untersuchten Zelllinien zeigten deutliche Unterschiede in Bezug auf ihre Uhr Phänotypen mit globalen Konsequenzen auf oszillierende Gene und Stoffwechselwege. Die Runterregulation des Uhrgens Bmal1 führte in SW480 Zellen zu einem metastatischen Phänotyp, der stark dem von SW620 Wildtypzellen ähnelte. Darüber hinaus führte die Runterregulation von Bmal1 zu einer Veränderung des metabolischen Phänotyps und zu einer modifizierten Antwort auf die Behandlung mit einem Glykolyseinhibitor. Die in diesem Projekt erzielten Ergebnisse unterstützen die postulierte Rolle von Bmal1 als Tumorsuppressor und verdeutlichen das reziproke Wechselspiel zwischen der zirkadianen Uhr und dem Stoffwechsel von Krebszellen und zeigen mögliche Auswirkungen einer deregulierten Uhr auf den Zellmetabolismus während der Tumorentwicklung. The circadian clock is an internal timing system that allows the entrainment of physiological and behavioural processes to the geophysical time with a periodicity of about 24 hours. It consists of a central pacemaker and peripheral clocks in every cell. In mammals, a distinct set of genes is interconnected in regulatory feedback loops, thereby generating oscillations in gene expression in the core-clock itself as well as in many target genes. Clock target genes are, among others, involved in cellular processes connected to tumour development and progression, including metabolic pathways, drug response pathways and the cell cycle. Malfunctions of the circadian clock are associated with different pathologies including cancer. The aim of this project was to study the role of the circadian clock in tumour development and progression with a focus on cancer metabolism and treatment response. The role of a deregulated clock was investigated in SW480 cells derived from a primary tumour and SW620 cells derived from a lymph node metastasis of the same patient. The investigated cell lines showed clear differences with respect to their clock phenotypes with consequences on global oscillating gene expression and alterations in metabolic pathways. A knockdown of the core-clock gene Bmal1 in SW480 cells induced a metastatic phenotype similar to SW620 wild type cells, as indicated by faster proliferation, lower apoptosis rate and a highly energetic metabolic phenotype. Furthermore, Bmal1-KD induced metabolic phenotype rewiring as seen by altered glycolytic activity and mitochondrial respiration and modified treatment response to metabolism-targeting anticancer treatment. The results obtained in this project reinforce the postulated role of Bmal1 as a tumour suppressor and elucidate a reciprocal interplay between the circadian clock and cancer metabolism with implications in metabolic phenotype rewiring during tumour progression.

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., Highlights • Primary and metastatic colon cancer cells from the same patient show differential time-dependent metabolic profiles. • Perturbations of the circadian clock induce differential alterations at the transcriptome level including metabolic pathways. • Core-clock gene (BMAL1) knockdown affects metabolic activity of cancer cells and impinges on treatment response. An evolutionary conserved molecular clockwork allows organisms to adapt physiology and biological processes to the geophysical time by driving various cellular pathways including metabolism and the cell cycle. In addition to dramatic metabolic alterations, cancer cells show severe changes in the biological clock likely to affect tumor progression and treatment response. Increasing efforts have been made to elucidate the connection between the circadian clock circuitry, tumor progression and cancer-associated metabolic alterations, yet a more detailed knowledge of this interplay is still missing. In this study, we investigated distinctive time-related transcriptomic and metabolic features in a cellular model of colon cancer progression and patient samples. We show that a disrupted biological clock leads to altered temporal profiles of gene expression, metabolic reprogramming and changes in drug response, and identify the hexokinase HKDC1 as a crucial element in this connection. Our results provide novel evidence regarding the complex interplay between the circadian clock and metabolic alterations in carcinogenesis and pave the way to a possible optimization of cancer treatment.