Anti-tumoral effect of desmethylclomipramine in lung cancer stem cells

Lung cancer is the most feared of all cancers because of its heterogeneity and resistance to available treatments. Cancer stem cells (CSCs) are the cell population responsible for lung cancer chemoresistance and are a very good model for testing new targeted therapies. Clomipramine is an FDA-approved antidepressant drug, able to inhibit in vitro the E3 ubiquitin ligase Itch and potentiate the pro-apoptotic effects of DNA damaging induced agents in several cancer cell lines. Here, we investigated the potential therapeutic effect of desmethylclomipramine (DCMI), the active metabolite of Clomipramine, on the CSCs homeostasis. We show that DCMI inhibits lung CSCs growth, decreases their stemness potential and increases the cytotoxic effect of conventional chemotherapeutic drugs. Being DCMI an inhibitor of the E3 ubiquitin ligase Itch, we also verified the effect of Itch deregulation on CSCs survival. We found that the siRNA-mediated depletion of Itch induces similar anti-proliferative effects on lung CSCs, suggesting that DCMI might exert its effect, at least in part, by inhibiting Itch. Notably, Itch expression is a negative prognostic factor in two primary lung tumors datasets, supporting the potential clinical relevance of Itch inhibition to circumvent drug resistance in the treatment of lung cancer

Enabling planetary science across light-years. Ariel Definition Study Report

Ariel, the Atmospheric Remote-sensing Infrared Exoplanet Large-survey, was adopted as the fourth medium-class mission in ESA's Cosmic Vision programme to be launched in 2029. During its 4-year mission, Ariel will study what exoplanets are made of, how they formed and how they evolve, by surveying a diverse sample of about 1000 extrasolar planets, simultaneously in visible and infrared wavelengths. It is the first mission dedicated to measuring the chemical composition and thermal structures of hundreds of transiting exoplanets, enabling planetary science far beyond the boundaries of the Solar System. The payload consists of an off-axis Cassegrain telescope (primary mirror 1100 mm x 730 mm ellipse) and two separate instruments (FGS and AIRS) covering simultaneously 0.5-7.8 micron spectral range. The satellite is best placed into an L2 orbit to maximise the thermal stability and the field of regard. The payload module is passively cooled via a series of V-Groove radiators; the detectors for the AIRS are the only items that require active cooling via an active Ne JT cooler. The Ariel payload is developed by a consortium of more than 50 institutes from 16 ESA countries, which include the UK, France, Italy, Belgium, Poland, Spain, Austria, Denmark, Ireland, Portugal, Czech Republic, Hungary, the Netherlands, Sweden, Norway, Estonia, and a NASA contribution

Two novel p63 transcriptional target genes regulating cell metabolism and metastasis

La famiglia di p53, costituita dai tre fattori di trascrizione p53, p63 e p73, assume un ruolo chiave nel direzionare il destino di una cellula verso la sopravvivenza o attivando meccanismi di morte cellulare programmata. Sebbene la loro struttura genica sia simile, p63 e p73 codificano per varie isoforme generate a partire da promotori diversi all’estremità 5’ e da meccanismi di splicing alternativo al 3’, a seguito del quale derivano isoforme TA complete del dominio di trans-attivazione all’N-terminale o isoforme tronche denominate ΔN. Il capostipite della famiglia, p53, è principalmente descritto come il guardiano del genoma ed è noto agire come soppressore dei tumori, essendo mutato in più del 50% dei tumori umani. Nell’ultimo decennio, l’identificazione di nuovi geni bersaglio di p53 implicati nella regolazione della glicolisi, come SCO2 e PGM, e nella difesa antiossidante come TIGAR, sestrine, ALDH4 e GPX1 sottolinea l’importanza di p53 nella regolazione del metabolismo energetico e dello stress ossidativo. Inoltre, recentemente, l’altro membro della famiglia, p73, è stato mostrato essere coinvolto nel metabolismo energetico attraverso il controllo trascrizionale del gene COX4A. Il fattore di trascrizione p63 svolge un ruolo chiave nello sviluppo e nel mantenimento dei tessuti epidermici e di tutti quelli di origine epiteliale. Nonostante p63 condivida alcuni ruoli biologici con gli altri membri della famiglia, come la regolazione del ciclo cellulare o della morte cellulare, il suo ruolo in alcuni processi cellulari basilari come il metabolismo energetico o in alcuni stadi patologici, come nei processi tumorigenici o metastatici non è stato ancora chiarito. In questo studio abbiamo dimostrato il coinvolgimento di p63 nei meccanismi di regolazione di due nuovi geni bersaglio: la glutaminasi mitocondriale 2 (GLS2) e il gene soppressore delle metastasi (MTSS1). La glutaminasi GLS2 idrolizza la glutammina in glutammato, un importante metabolita coinvolto nella via energetica di sintesi dell’ATP e nei processi di difesa cellulare contro le specie ossidanti. Abbiamo dimostrato che l’espressione di GLS2 aumenta in seguito all’espressione ectopica del’isoforma TAp63 in varie linee cellulari. Tale aumento è confermato anche dall’induzione endogena di TAp63 in seguito all’inibizione delle deacetilasi istoniche. Attraverso saggi di immunoprecipitazione della cromatina e saggi della luciferasi è stato confermato il legame diretto di TAp63 sulla sequenza di DNA consenso per p53/p63 localizzata all’interno del promotore di GLS2. Abbiamo inoltre osservato che l’espressione di GLS2 e di TAp63 aumenta contemporaneamente sia durante il differenziamento in vitro di cheratinociti primari umani, e sia in cellule tumorali esposte a stress ossidativo. I nostri risultati dimostrano che la deplezione di GLS2, da una parte inibisce il differenziamento dei cheratinociti umani in vitro e, dall’altra, aumenta la morte cellulare indotta da ROS in cellule tumorali. Questi dati mostrano che l’asse TAp63/GLS2 può essere funzionalmente importante sia nei processi fisiologici che patologici, rappresentando una via cellulare anti-ossidante in grado di compensare l’attività di p53 quando assente. Infatti, l’espressione di p63 correla con quella di GLS2 nei tumori umani del carcinoma del colon e nei carcinomi a cellule squamose della testa e del collo, suggerendo che in alcuni tumori questo sistema potrebbe essere importante nel proteggere le cellule tumorali dallo stress ossidativo. Un altro gene che abbiamo identificato essere regolato da p63 è MTSS1. Il gene MTSS1 è stato originariamente identificato come soppressore delle metastasi, la cui espressione era assente nel carcinoma metastatico della vescica e della prostata. Tuttavia, recentemente è stato dimostrato che la proteina MTSS1 è in grado di agire come un oncogene e come un fattore pro-migratorio nel melanoma. I nostri risultati caratterizzano il gene MTSS1 come un nuovo bersaglio trascrizionale di p63, implicato nella regolazione dei processi di migrazione e invasione cellulare. Abbiamo trovato che sia in cellule normali che in cellule tumorali, l’espressione ectopica di p63, ed in particolare delle isoforme di ΔNp63, è in grado di guidare l’espressione di MTSS1. Inoltre abbiamo confermato che ques11ta regolazione è possibile grazie al legame diretto di p63 a un elemento di risposta nel DNA, avente un motivo di sequenza specifico per p63 e localizzato nelle regioni introniche del locus genico di MTSS1. Inoltre, da un punto di vista biologico, l’espressione di MTSS1 ΔNp63-dipendente, sembra regolare le proprietà migratorie e invasive di cellule tumorali. Da sottolineare, i nostri dati indicano che l’espressione di MTSS1 correla positivamente con quella di ΔNp63 in 3 datasets di tumori alla mammella, alla vescica e alla prostata. Inoltre, in tre datasets di tumori umani della mammella, la co-espressione di MTSS1 e p63 è un fattore prognostico negativo per la sopravvivenza del paziente, suggerendo che questo asse p63/MTSS1 possa essere funzionalmente importante nel regolare la progressione del tumore. In conclusione, attraverso l’individuazione e la caratterizzazione di due nuovi geni bersaglio di p63, abbiamo sottolineato l’importanza di p63 in due processi funzionalmente interconnessi quali il metabolismo cellulare e i processi di migrazione e invasione cellulare, che sono a loro volta fondamentali per la corretta regolazione della proliferazione e della sopravvivenza cellulare sia in sistemi fisiologici che patologici.The transcription factor p63 is critical for the development and maintenance of epidermal tissues and its role in other processes such as cell energy metabolism, or in pathological conditions, including tumorigenesis and metastasis, is far to be completely characterized. Here, we report the characterization of two novel p63 target genes: the mitochondrial Glutaminase 2 (GLS2), and the Metastasis Suppressor 1 (MTSS1) gene. GLS2 converts glutamine into glutamate, an important metabolite involved in ATP production and anti-oxidant cellular defense. We found that the p63 TA-isoforms induce GLS2 expression in several cell lines and, by ChIP analysis and luciferase assay, we demonstrated the direct binding of TAp63 to the p53/p63 consensus DNA responsive sequence localized within the GLS2 promoter. GLS2 and TAp63 expression concomitantly increases both during the in vitro differentiation of primary human keratinocytes, and in cancer cells exposed to oxidative stresses, while depletion of GLS2 inhibits skin differentiation and sensitizes tumor cells to ROS-induced apoptosis. Our findings show that TAp63/GLS2 axis may be functionally important as a cellular anti-oxidant pathway in the absence of p53 for both physiological and pathological processes. Metastasis Suppressor 1 (MTSS1) was originally identified as a metastasis suppressor protein whose expression is missing in metastatic bladder carcinoma and prostate cancer. However, recent findings indicate that MTSS1 acts as oncogene and pro-migratory factor in melanoma tumors. Here, we characterized MTSS1 as a new p63 transcriptional target gene involved in the regulation of the migration/invasion processes. We found that in normal and in cancer cell lines p63 is able to drive the expression of MTSS1 through binding to one p63 binding element localized in the MTSS1 locus. We also found that the p63-mediated expression of MTSS1 plays a critical role in the regulation of the migration and invasion properties of tumor cells. Importantly, the expression of MTSS1 positively correlates with that of ΔNp63 isoforms in breast, bladder and prostate human tumors datasets. Furthermore, in three human breast tumors datasets the MTSS1-p63 co-expression is a negative prognostic factor on patient survival, suggesting that the MTSS1/p63 axis might be functionally important to regulate tumor progression. In conclusion, by identifying and characterizing two novel p63 target genes, we have unveiled the importance of p63 in two functionally interconnected pathways, cell metabolism and migration/invasion processes, which are critical for the proper regulation of cell proliferation and survival in both physiological and pathological process

A MYC and RAS co-activation signature in localized prostate cancer drives bone metastasis and castration resistance

Understanding the intricacies of lethal prostate cancer poses specific challenges due to difficulties in accurate modeling of metastasis in vivo. Here we show that NPK(EYFP) mice (for Nkx3.1(CreERT2/+); Pten(flox/flox); Kras(LSL-G12D/+); R26R-CAG-(LSL-EYFP/+)) develop prostate cancer with a high penetrance of metastasis to bone, thereby enabling detection and tracking of bone metastasis in vivo and ex vivo. Transcriptomic and whole-exome analyses of bone metastasis from these mice revealed distinct molecular profiles conserved between human and mouse and specific patterns of subclonal branching from the primary tumor. Integrating bulk and single-cell transcriptomic data from mouse and human datasets with functional studies in vivo unravels a unique MYC/RAS co-activation signature associated with prostate cancer metastasis. Finally, we identify a gene signature with prognostic value for time to metastasis and predictive of treatment response in human patients undergoing androgen receptor therapy across clinical cohorts, thus uncovering conserved mechanisms of metastasis with potential translational significance

Ariel: Enabling planetary science across light-years

Ariel Definition Study ReportAriel Definition Study Report, 147 pages. Reviewed by ESA Science Advisory Structure in November 2020. Original document available at: https://www.cosmos.esa.int/documents/1783156/3267291/Ariel_RedBook_Nov2020.pdf/Ariel, the Atmospheric Remote-sensing Infrared Exoplanet Large-survey, was adopted as the fourth medium-class mission in ESA's Cosmic Vision programme to be launched in 2029. During its 4-year mission, Ariel will study what exoplanets are made of, how they formed and how they evolve, by surveying a diverse sample of about 1000 extrasolar planets, simultaneously in visible and infrared wavelengths. It is the first mission dedicated to measuring the chemical composition and thermal structures of hundreds of transiting exoplanets, enabling planetary science far beyond the boundaries of the Solar System. The payload consists of an off-axis Cassegrain telescope (primary mirror 1100 mm x 730 mm ellipse) and two separate instruments (FGS and AIRS) covering simultaneously 0.5-7.8 micron spectral range. The satellite is best placed into an L2 orbit to maximise the thermal stability and the field of regard. The payload module is passively cooled via a series of V-Groove radiators; the detectors for the AIRS are the only items that require active cooling via an active Ne JT cooler. The Ariel payload is developed by a consortium of more than 50 institutes from 16 ESA countries, which include the UK, France, Italy, Belgium, Poland, Spain, Austria, Denmark, Ireland, Portugal, Czech Republic, Hungary, the Netherlands, Sweden, Norway, Estonia, and a NASA contribution

Ariel: Enabling planetary science across light-years

Ariel, the Atmospheric Remote-sensing Infrared Exoplanet Large-survey, was adopted as the fourth medium-class mission in ESA's Cosmic Vision programme to be launched in 2029. During its 4-year mission, Ariel will study what exoplanets are made of, how they formed and how they evolve, by surveying a diverse sample of about 1000 extrasolar planets, simultaneously in visible and infrared wavelengths. It is the first mission dedicated to measuring the chemical composition and thermal structures of hundreds of transiting exoplanets, enabling planetary science far beyond the boundaries of the Solar System. The payload consists of an off-axis Cassegrain telescope (primary mirror 1100 mm x 730 mm ellipse) and two separate instruments (FGS and AIRS) covering simultaneously 0.5-7.8 micron spectral range. The satellite is best placed into an L2 orbit to maximise the thermal stability and the field of regard. The payload module is passively cooled via a series of V-Groove radiators; the detectors for the AIRS are the only items that require active cooling via an active Ne JT cooler. The Ariel payload is developed by a consortium of more than 50 institutes from 16 ESA countries, which include the UK, France, Italy, Belgium, Poland, Spain, Austria, Denmark, Ireland, Portugal, Czech Republic, Hungary, the Netherlands, Sweden, Norway, Estonia, and a NASA contribution