Circadian Dysregulation of the TGFβ/SMAD4 Pathway Modulates Metastatic Properties and Cell Fate Decisions in Pancreatic Cancer Cells

Impairment of circadian rhythms impacts carcinogenesis. SMAD4, a clock-controlled gene and central component of the TGFβ canonical pathway, is frequently mutated in pancreatic ductal adenocarcinoma (PDA), leading to decreased survival. Here, we used an in vitro PDA model of SMAD4-positive and SMAD4-negative cells to investigate the interplay between circadian rhythms, the TGFβ canonical signaling pathway, and its impact on tumor malignancy. Our data show that TGFβ1, SMAD3, SMAD4, and SMAD7 oscillate in a circadian fashion in SMAD4-positive PDA cells, whereas altering the clock impairs the mRNA dynamics of these genes. Furthermore, the expression of the clock genes DEC1, DEC2, and CRY1 varied depending on SMAD4 status. TGFβ pathway activation resulted in an altered clock, cell-cycle arrest, accelerated apoptosis rate, enhanced invasiveness, and chemosensitivity. Our data suggest that the impact of TGFβ on the clock is SMAD4-dependent, and S MAD3, SMAD4, DEC1, and CRY1 involved in this cross-talk affect PDA patient survival

Analysis of the Circadian Regulation of Cancer Hallmarks by a Cross-Platform Study of Colorectal Cancer Time-Series Data Reveals an Association with Genes Involved in Huntington's Disease

Accumulating evidence points to a link between circadian clock dysfunction and the molecular events that drive tumorigenesis. Here, we investigated the connection between the circadian clock and the hallmarks of cancer in an in vitro model of colorectal cancer (CRC). We used a cross-platform data normalization method to concatenate and compare available microarray and RNA-sequencing time series data of CRC cell lines derived from the same patient at different disease stages. Our data analysis suggests differential regulation of molecular pathways between the CRC cells and identifies several of the circadian and likely clock-controlled genes (CCGs) as cancer hallmarks and circadian drug targets. Notably, we found links of the CCGs to Huntington's disease (HD) in the metastasis-derived cells. We then investigated the impact of perturbations of our candidate genes in a cohort of 439 patients with colon adenocarcinoma retrieved from the Cancer Genome Atlas (TCGA). The analysis revealed a correlation of the differential expression levels of the candidate genes with the survival of patients. Thus, our study provides a bioinformatics workflow that allows for a comprehensive analysis of circadian properties at different stages of colorectal cancer, and identifies a new association between cancer and HD

It’s About Time: The Circadian Network as Time-Keeper for Cognitive Functioning, Locomotor Activity and Mental Health

A variety of organisms including mammals have evolved a 24h, self-sustained timekeeping machinery known as the circadian clock (biological clock), which enables to anticipate, respond, and adapt to environmental influences such as the daily light and dark cycles. Proper functioning of the clock plays a pivotal role in the temporal regulation of a wide range of cellular, physiological, and behavioural processes. The disruption of circadian rhythms was found to be associated with the onset and progression of several pathologies including sleep and mental disorders, cancer, and neurodegeneration. Thus, the role of the circadian clock in health and disease, and its clinical applications, have gained increasing attention, but the exact mechanisms underlying temporal regulation require further work and the integration of evidence from different research fields. In this review, we address the current knowledge regarding the functioning of molecular circuits as generators of circadian rhythms and the essential role of circadian synchrony in a healthy organism. In particular, we discuss the role of circadian regulation in the context of behaviour and cognitive functioning, delineating how the loss of this tight interplay is linked to pathological development with a focus on mental disorders and neurodegeneration. We further describe emerging new aspects on the link between the circadian clock and physical exercise-induced cognitive functioning, and its current usage as circadian activator with a positive impact in delaying the progression of certain pathologies including neurodegeneration and brain-related disorders. Finally, we discuss recent epidemiological evidence pointing to an important role of the circadian clock in mental health.Peer Reviewe

A comprehensive bioinformatic analysis of circadian regulation of cellular pathways in an in-vitro model of colorectal cancer (CRC) and neurodegeneration

At the heart of the universe lies the synchrony of nature at every scale, including in our bodies. Building upon this concept Nobel Prize in Physiology or Medicine in 2017 was awarded to the discovery of the circadian clock which is the fascinating molecular genetic mechanism that allows organisms to anticipate, respond, adjust, and synchronize with recurring periodic events. The human circadian system, which is essential for fine-tuning the timing of biological events around the 24-hour solar day, comprises a network of clocks distributed throughout the body, with the suprachiasmatic nucleus in the brain acting as a central regulator. Any disruption to cellular clocks or disruption of their synchrony can lead to alterations in cellular pathways such as cell cycle or DNA repair. This doctoral dissertation aimed to characterize expression changes at the transcriptomic level upon disruption of core-clock genes on common cellular pathways associated with cancer and neurodegeneration (ND). High-throughput data sets retrieved from idiopathic Parkinson's disease (IPD) patients (438 samples) and an in vitro colorectal carcinoma (CRC), including Wild Type (WT) and clock Knock Out (KO) cells (64 samples) were analyzed. The gene expression changes observed in individuals with IPD were similar to the patterns seen in core-clock KO cells. Curated literature and KEGG pathways searches were used to determine a set of relevant pathways and their corresponding involved genes in ND and cancer. Among those genes we found genes associated with genetic forms of PD such as SNCA, GBA, and core-clock regulator DBP. Notably, co-expression of core-clock network was altered in IPD patients compared to healthy controls, pointing to variations in the expression profile of PD-associated genes that may result from the disruption of the core-clock (as observed in vitro in the CRC model). Additionally, our subsequent analysis using the cancer genome atlas (TCGA) database revealed that genes, identified in our analysis, belong to cancer hallmark pathways and had a significant impact on clinical traits such as overall survival in colon cancer patients. Researchers have just started to shed light on the consequences of the disruptions in circadian rhythms, which could contribute to a deeper understanding of the molecular mechanisms underlying pathological development. Our results further point to a common regulation of specific pathways in cancer and ND and highlight the importance of the circadian system in this regulation. By characterizing these oscillations, an overlooked feature in the clinical routines, may actually reveal molecular signatures that can improve the diagnosis and monitoring of these pathologies.Dem Universum liegt eine Synchronität der Natur in jedem Bereich zugrunde, auch dem menschlichen Körper. Basierend auf diesem Konzept wurde die Entdeckung des faszinierenden molekulargenetischen Mechanismus der zirkadianen Uhr, mit dem Nobelpreis im Jahr 2017 gewürdigt. Diese ermöglicht Organismen periodisch wiederkehrende Prozesse zu antizipieren, darauf zu reagieren, sich anzupassen und zu synchronisieren. Das zirkadiane System des Menschen, das für die zeitliche Anpassung biologischer Ereignisse an den 24-Stunden-Tag notwendig ist, besteht aus einem Netzwerk molekularer Uhren, welches im ganzen Körper verteilt ist und in dem der suprachiasmatische Nukleus im Gehirn als Hauptregulator funktioniert. Jede Störung der zellulären Uhren oder ihrer Synchronität kann zu Veränderungen zellulärer Signalwege wie z.B. dem Zellzyklus oder der DNA-Reparatur führen. Ziel dieser Dissertation war es, Veränderungen herkömmlicher zellulärer Prozesse auf Transkriptom-Ebene zu charakterisieren, welche durch Störung des zirkadianen Systems auftreten und mit Krebs und Neurodegeneration (ND) in Verbindung stehen. Zu diesem Zweck wurden Hochdurchsatzdaten von Patienten mit idiopathischer Parkinson-Krankheit (IPD) (438 Proben), sowie von einem in-vitro-Modell des kolorektalen Karzinoms (CRC), einschließlich Wildtyp (WT)- und zirkadiane Knock-Out (KO)-Zellen, (64 Proben) analysiert. Mehrere Genexpressionsveränderungen der IPD-Patienten ähnelten Expressionsprofilen der CRC zirkadianen KO-Zellen. Mithilfe einer kuratierten Literatur- und KEGG-Suche wurden eine Reihe relevanter Signalwege und die daran beteiligten Gene bei ND und Krebs ermittelt. Einige dieser Gene sind dafür bekannt mit einer genetischen Formen der Parkinson-Krankheit in Verbindung zu stehen, z.B. SNCA, GBA und das Core-Clock-Gen DBP. Im Vergleich zu gesunden Kontrollpersonen, wurden bei Parkinson-Patienten insbesondere Veränderungen des Core-Clock-Netzwerks festgestellt. Dies weist darauf hin, dass Variationen der Expressionsprofile Parkinson-assoziierter Gene möglicherweise aus der Störung von Core-Clock-Genen resultieren (wie in-vitro im CRC-Modell beobachtet). Darüber hinaus ergab unsere anschließende Analyse mit Hilfe der The Cancer-Genome Atlas (TCGA) Datenbank, dass die identifizierten Gene zu Krebs-Hallmark-Signalwegen gehören und einen signifikanten Einfluss auf klinische Eigenschaften wie die Überlebensdauer bei Darmkrebspatienten haben. Als Wissenschaftler stehen wir noch am Anfang klinische Auswirkungen von Störungen des zirkadianen Systems zu erforschen - in Zukunft wird dies jedoch zu einem besseren Verständnis molekularer Mechanismen von Krankheitsentwicklung beitragen. Unsere Ergebnisse stützen die Hypothese einer gemeinsamen Regulierung bestimmter Signalwege bei Krebs und ND und unterstreichen die dabei wichtige Rolle des zirkadianen Systems. Da die Rolle zirkadianer Rhythmik in der klinischen Routine noch oft übersehen wird, könnte deren Charakterisierung durch Identifikation entsprechender molekular Signaturen tatsächlich die Diagnose und Überwachung dieser Pathologien verbessern

Sex and age-dependent characterization of the circadian clock as a potential biomarker for physical performance: A prospective study protocol.

IntroductionCircadian rhythms (CR) regulate daily cycles in behavior, physiology and molecular processes. CRs are endogenous and vary across individuals. Seasonal changes can influence CR. Accordingly, rhythms with different characteristics (amplitude, phase) are depicted during the summer months, as compared to winter. Increasing evidence points to an influence of circadian regulation on physical performance. Here, we aim to obtain a comprehensive circadian gene expression profile for physically active individuals, which can potentially be used for the identification of optimal time intervals for physical exercise.Methods and analysisTo explore these different aspects, we propose a study where we will carry out a molecular analysis of CR by measuring the expression of specific clock and clock-controlled genes, based on a non-invasive approach using RNA extracted from saliva in physically active, healthy participants. We will collect data across two seasons and use computational algorithms to integrate the molecular data with hormonal data (cortisol and melatonin), and generate a profile of CR in healthy individuals of different sex and age groups. Finally, we will use computational tools to predict optimal time intervals for physical performance based on the above-described data, thereby retrieving valuable data on the circadian clock as a key factor for health maintenance and optimization

A computational analysis in a cohort of Parkinson’s disease patients and clock-modified colorectal cancer cells reveals common expression alterations in clock-regulated genes

© 2021 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).Increasing evidence suggests a role for circadian dysregulation in prompting disease-related phenotypes in mammals. Cancer and neurodegenerative disorders are two aging related diseases reported to be associated with circadian disruption. In this study, we investigated a possible effect of circadian disruption in Parkinson's disease (PD) and colorectal cancer (CRC). We used high-throughput data sets retrieved from whole blood of idiopathic PD (IPD) patients and time course data sets derived from an in vitro model of CRC including the wildtype and three core-clock knockout (KO) cell lines. Several gene expression alterations in IPD patients resembled the expression profiles in the core-clock KO cells. These include expression changes in DBP, GBA, TEF, SNCA, SERPINA1 and TGFB1. Notably, our results pointed to alterations in the core-clock network in IPD patients when compared to healthy controls and revealed variations in the expression profile of PD-associated genes (e.g., HRAS and GBA) upon disruption of the core-clock genes. Our study characterizes changes at the transcriptomic level following circadian clock disruption on common cellular pathways associated with cancer and neurodegeneration (e.g., immune system, energy metabolism and RNA processing), and it points to a significant influence on the overall survival of colon cancer patients for several genes resulting from our analysis (e.g., TUBB6, PAK6, SLC11A1).info:eu-repo/semantics/publishedVersio

Skeletal muscle gene expression dysregulation in long-term spaceflights and aging is clock-dependent

Abstract The circadian clock regulates cellular and molecular processes in mammals across all tissues including skeletal muscle, one of the largest organs in the human body. Dysregulated circadian rhythms are characteristic of aging and crewed spaceflight, associated with, for example, musculoskeletal atrophy. Molecular insights into spaceflight-related alterations of circadian regulation in skeletal muscle are still missing. Here, we investigated potential functional consequences of clock disruptions on skeletal muscle using published omics datasets obtained from spaceflights and other clock-altering, external (fasting and exercise), or internal (aging) conditions on Earth. Our analysis identified alterations of the clock network and skeletal muscle-associated pathways, as a result of spaceflight duration in mice, which resembles aging-related gene expression changes observed in humans on Earth (e.g., ATF4 downregulation, associated with muscle atrophy). Furthermore, according to our results, external factors such as exercise or fasting lead to molecular changes in the core-clock network, which may compensate for the circadian disruption observed during spaceflights. Thus, maintaining circadian functioning is crucial to ameliorate unphysiological alterations and musculoskeletal atrophy reported among astronauts