Clonal chromosomal mosaicism and loss of chromosome Y in elderly men increase vulnerability for SARS-CoV-2

The pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2, COVID-19) had an estimated overall case fatality ratio of 1.38% (pre-vaccination), being 53% higher in males and increasing exponentially with age. Among 9578 individuals diagnosed with COVID-19 in the SCOURGE study, we found 133 cases (1.42%) with detectable clonal mosaicism for chromosome alterations (mCA) and 226 males (5.08%) with acquired loss of chromosome Y (LOY). Individuals with clonal mosaic events (mCA and/or LOY) showed a 54% increase in the risk of COVID-19 lethality. LOY is associated with transcriptomic biomarkers of immune dysfunction, pro-coagulation activity and cardiovascular risk. Interferon-induced genes involved in the initial immune response to SARS-CoV-2 are also down-regulated in LOY. Thus, mCA and LOY underlie at least part of the sex-biased severity and mortality of COVID-19 in aging patients. Given its potential therapeutic and prognostic relevance, evaluation of clonal mosaicism should be implemented as biomarker of COVID-19 severity in elderly people. Among 9578 individuals diagnosed with COVID-19 in the SCOURGE study, individuals with clonal mosaic events (clonal mosaicism for chromosome alterations and/or loss of chromosome Y) showed an increased risk of COVID-19 lethality

Los factores de transcripción TBX15 e YY1 en cáncer. Función y regulación de TBX15. Expresión de YY1 en cáncer de tiroides

TBX15 e YY1 son dos factores de transcripción y los factores de transcripción, como moléculas transductoras de señales, ejercen un papel clave en la regulación de muchos procesos básicos del funcionamiento y fisiología de la célula, como pueden ser la proliferación celular, la inducción de la apoptosis o la reparación del DNA. Por lo tanto, la expresión y función aberrantes de los factores de transcripción son un punto importante en la aparición y desarrollo del cáncer. Los factores de transcripción en cáncer actúan como oncogenes o genes supresores de tumores y su expresión se encuentra alterada en muchos tipos de cáncer. En cáncer de tiroides se ha descrito la asociación de ciertos factores de transcripción específicos del tiroides con este tipo de cáncer, pero no existe información acerca de la implicación de otros factores de transcripción generales, ni tampoco sobre TBX15 e YY1. La implicación de YY1 en cáncer está documentada; sin embargo, no existen estudios relativos a la posible implicación de TBX15 en cáncer. En este contexto, la presente tesis aporta conocimiento sobre el papel del factor de transcripción TBX15 en el desarrollo del cáncer, y se analiza la expresión de YY1 en cáncer de tiroides. Nuestro estudio describe una nueva función del factor de transcripción TBX15 como inhibidor de la apoptosis celular, lo que puede contribuir al potencial proliferativo de las células cancerígenas y sugiere a TBX15 como una diana terapéutica potencial en el tratamiento del cáncer. También, hemos demostrado que NFkB regula positivamente la transcripción de TBX15 mediante su unión a una región reguladora en la zona 5’-distal del gen TBX15. La relación entre TBX15 y NFkB puede ser importante para entender el papel de TBX15 en el cáncer. Con referencia al factor de transcripción YY1, nuestros resultados representan el primer estudio sobre la implicación de YY1 en el cáncer de tiroides sin tener información previa sobre la expresión de este factor en este tipo cáncer. Mostramos como YY1 se encuentra sobreexpresado en cáncer diferenciado de tiroides, siendo más frecuente su expresión positiva en el tipo papilar que en el folicular, poniendo en evidencia la posible implicación de YY1 en el cáncer de tiroides.TBX15 and YY1 are transcription factors; these molecules are able to transduction signals, being essential in the regulation of many basic cellular processes including cell proliferation and apoptosis. Therefore, the anomalous expression and function of these transcription factors is crucial in the beginning and in the development of cancer. Transcription factors act as oncogenes or tumor suppressor genes and their expression is found altered in multiple types of cancer. Specific transcription factors of the thyroid gland have been reported to be associated with thyroid cancer; however there is no information about the implication of general transcription factors, such as TBX15 or YY1. The involvement of YY1 in cancer is well documented; whereas there are scarcely any studies describing the possible implication of TBX15 in cancer. In this context, the present thesis provides knowledge about the role of transcription factor TBX15 in the development of cancer; moreover, it also analyzes the expression of transcription factor YY1 in differentiated thyroid cancer. Our study reveals a novel function of transcription factor TBX15 as an inhibitor of cellular apoptosis, which can contribute to the proliferative potential of cancer cells, and may suggest TBX15 as a potential therapeutic target in cancer treatment. Furthermore, we have also proven that NFkB activates the transcription of TBX15 by binding to the 5’-flanking regulatory region of the gene TBX15. Thus, the interaction between TBX15 and NFkB could prove to be important to understand the function of TBX15 in cancer. Without any previous information regarding the expression of transcription factor YY1 in thyroid cancer, our results represent the first study about the implication of YY1 in this type of cancer. We demonstrate how YY1 is overexpressed in differentiated thyroid cancer, and what’s more, its positive expression has been found to be more frequent in the papillary type rather than in the follicular type. Therefore these results evidence the possible implication of transcription factor YY1 in thyroid cancer

Los Factores de transcripción TBX15 e YY1 en cáncer : función y regulación de TBX15: expresión de YY1 en cáncer de tiroides /

TBX15 e YY1 son dos factores de transcripción y los factores de transcripción, como moléculas transductoras de señales, ejercen un papel clave en la regulación de muchos procesos básicos del funcionamiento y fisiología de la célula, como pueden ser la proliferación celular, la inducción de la apoptosis o la reparación del DNA. Por lo tanto, la expresión y función aberrantes de los factores de transcripción son un punto importante en la aparición y desarrollo del cáncer. Los factores de transcripción en cáncer actúan como oncogenes o genes supresores de tumores y su expresión se encuentra alterada en muchos tipos de cáncer. En cáncer de tiroides se ha descrito la asociación de ciertos factores de transcripción específicos del tiroides con este tipo de cáncer, pero no existe información acerca de la implicación de otros factores de transcripción generales, ni tampoco sobre TBX15 e YY1. La implicación de YY1 en cáncer está documentada; sin embargo, no existen estudios relativos a la posible implicación de TBX15 en cáncer. En este contexto, la presente tesis aporta conocimiento sobre el papel del factor de transcripción TBX15 en el desarrollo del cáncer, y se analiza la expresión de YY1 en cáncer de tiroides. Nuestro estudio describe una nueva función del factor de transcripción TBX15 como inhibidor de la apoptosis celular, lo que puede contribuir al potencial proliferativo de las células cancerígenas y sugiere a TBX15 como una diana terapéutica potencial en el tratamiento del cáncer. También, hemos demostrado que NFkB regula positivamente la transcripción de TBX15 mediante su unión a una región reguladora en la zona 5'-distal del gen TBX15. La relación entre TBX15 y NFkB puede ser importante para entender el papel de TBX15 en el cáncer. Con referencia al factor de transcripción YY1, nuestros resultados representan el primer estudio sobre la implicación de YY1 en el cáncer de tiroides sin tener información previa sobre la expresión de este factor en este tipo cáncer. Mostramos como YY1 se encuentra sobreexpresado en cáncer diferenciado de tiroides, siendo más frecuente su expresión positiva en el tipo papilar que en el folicular, poniendo en evidencia la posible implicación de YY1 en el cáncer de tiroides.TBX15 and YY1 are transcription factors; these molecules are able to transduction signals, being essential in the regulation of many basic cellular processes including cell proliferation and apoptosis. Therefore, the anomalous expression and function of these transcription factors is crucial in the beginning and in the development of cancer. Transcription factors act as oncogenes or tumor suppressor genes and their expression is found altered in multiple types of cancer. Specific transcription factors of the thyroid gland have been reported to be associated with thyroid cancer; however there is no information about the implication of general transcription factors, such as TBX15 or YY1. The involvement of YY1 in cancer is well documented; whereas there are scarcely any studies describing the possible implication of TBX15 in cancer. In this context, the present thesis provides knowledge about the role of transcription factor TBX15 in the development of cancer; moreover, it also analyzes the expression of transcription factor YY1 in differentiated thyroid cancer. Our study reveals a novel function of transcription factor TBX15 as an inhibitor of cellular apoptosis, which can contribute to the proliferative potential of cancer cells, and may suggest TBX15 as a potential therapeutic target in cancer treatment. Furthermore, we have also proven that NFkB activates the transcription of TBX15 by binding to the 5'-flanking regulatory region of the gene TBX15. Thus, the interaction between TBX15 and NFkB could prove to be important to understand the function of TBX15 in cancer. Without any previous information regarding the expression of transcription factor YY1 in thyroid cancer, our results represent the first study about the implication of YY1 in this type of cancer. We demonstrate how YY1 is overexpressed in differentiated thyroid cancer, and what's more, its positive expression has been found to be more frequent in the papillary type rather than in the follicular type. Therefore these results evidence the possible implication of transcription factor YY1 in thyroid cancer

Harmful effects of true-to-life nanoplastics derived from PET water bottles in human alveolar macrophages

Altres ajuts: acords transformatius de la UABThe increasing presence of secondary micro/nanoplastics (MNPLs) in the environment requires knowing if they represent a real health concern. To such end, an important point is to test representative MNPLs such as the denominated true-to-life MNPLs, resulting from the degradation of plastic goods in lab conditions. In this study, we have used polyethylene terephthalate (PET) NPLs resulting from the degradation of PET water bottles. Since inhalation is an important exposure route to environmental MNPLS, we have used mouse alveolar macrophages (MH-S) as a target cell, and the study focused only on the cells that have internalized them. This type of approach is novel as it may capture the realistic adverse effects of PETNPLs only in the internalized cells, thereby mitigating any biases while assessing the risk of these MNPLs. Furthermore, the study utilized a set of biomarkers including intracellular reactive oxygen species (ROS) levels, variations on the mitochondrial membrane potential values, and the macrophage polarization to M1 (pro-inflammatory response) and M2 (anti-proinflammatory response) as possible cellular effects due to PETNPLs in only the cells that internalized PETNPLs. After exposures lasting for 3 and 24 h to a range of concentrations (0, 25, 50, and 100 μg/mL) the results indicate that no toxicity was induced despite the 100% internalization observed at the highest concentration. Significant intracellular levels of ROS were observed, mainly at exposures lasting for 24 h, in an indirect concentration-effect relationship. Interestingly, a reduction in the mitochondrial membrane potential was observed, but only at exposures lasting for 24 h, but without a clear concentration-effect relationship. Finally, PETNPL exposure shows a significant polarization from M0 to M1 and M2 subtypes. Polarization to M1 (pro-inflammatory stage) was more marked and occurred at both exposure times. Polarization to M2 (anti-inflammatory stage) was only observed after exposures lasting for 24 h. Due to the relevance of the described biomarkers, our results underscore the need for further research, to better understand the health implications associated with MNPL exposure

NF-kB-mediated the expression of TBX15 in cancer cells

TBX15 is a T-box transcription factor essential for development, also proposed as a marker in prostate cancer; and, recently, its antiapoptotic function indicates a role in carcinogenesis. Regulation of TBX15 is uncovered. In this study, we investigated the regulation of TBX15 expression in human cancer cells, by analyzing the regulatory function of a 5'-distal conserved region of TBX15. Bisulfite sequencing showed high methylation of the CpG island contained in this region that was not correlated with TBX15 mRNA levels, in the cancer cell lines analyzed; however, after 5-aza-dC treatment of TPC-1 cells an increase of TBX15 expression was observed. We also found a significant response of TBX15 to TNF-α activation of the NF-κB pathway using five cancer cell lines, and similar results were obtained when NF-κB was activated with PMA/ionomycin. Next, by luciferase reporter assays, we identified the TBX15 regulatory region containing two functional NF-κB binding sites with response to NF-κBp65, mapping on the -3302 and -3059 positions of the TBX15 gene. Moreover, a direct interaction of NF-κBp65 with one of the two NF-κB binding sites was indicated by ChIP assays. In summary, we provide novel data showing that NF-κB signaling up-regulates TBX15 expression in cancer cells. Furthermore, the link between TBX15 and NF-κB found in this study may be important to understand cancer and development processes

NF-kB-mediated the expression of TBX15 in cancer cells

TBX15 is a T-box transcription factor essential for development, also proposed as a marker in prostate cancer; and, recently, its antiapoptotic function indicates a role in carcinogenesis. Regulation of TBX15 is uncovered. In this study, we investigated the regulation of TBX15 expression in human cancer cells, by analyzing the regulatory function of a 5'-distal conserved region of TBX15. Bisulfite sequencing showed high methylation of the CpG island contained in this region that was not correlated with TBX15 mRNA levels, in the cancer cell lines analyzed; however, after 5-aza-dC treatment of TPC-1 cells an increase of TBX15 expression was observed. We also found a significant response of TBX15 to TNF-α activation of the NF-κB pathway using five cancer cell lines, and similar results were obtained when NF-κB was activated with PMA/ionomycin. Next, by luciferase reporter assays, we identified the TBX15 regulatory region containing two functional NF-κB binding sites with response to NF-κBp65, mapping on the -3302 and -3059 positions of the TBX15 gene. Moreover, a direct interaction of NF-κBp65 with one of the two NF-κB binding sites was indicated by ChIP assays. In summary, we provide novel data showing that NF-κB signaling up-regulates TBX15 expression in cancer cells. Furthermore, the link between TBX15 and NF-κB found in this study may be important to understand cancer and development processes

Expression of <i>TBX15</i>mRNA in different cell lines after stimulation with TNF-α.

<p>(A) qRT-PCR of <i>TBX15</i>mRNA analysis in FRO, CGTH, TPC-1 and HeLa cells after 2h of TNF-α treatment. (B) qRT-PCR of <i>TBX15</i>mRNA (left) and <i>CXCL1</i>mRNA (right) analysis in p65^+/+ and p65^<i>-/-</i> MEF cells after TNF-α treatment. Data were normalized to <i>RPL27</i> and expressed as fold change referred to untreated cells whose <i>TBX15</i>mRNA or <i>CXCL1</i>mRNA relative expression was defined as 1. Data are mean±SD of mRNA levels of two independent experiments in triplicates. ** indicates p-value < 0.01.</p

Chromatin inmunoprecipitation (ChIP) assays.

<p>(A) The PCR amplified regions comprising the predicted NFκB binding sites (kBS1, kBS2 and kBS3) are indicated as rectangles, with location of these specific DNA segments indicated by numbers. (B) TPC and CGTH cells were treated with TNF-α and ChIP assays performed using IgG (mock) and NFκBp65 antibodies. Imnmunoprecipitated DNA was analyzed by qPCR with primers targeted to the predicted NFκB binding sites (kBS1, kBS2 and kBS3). The quantitative data reflect occupancy calculated as % input and represented as mean±SD of two independent experiments. *: P<0.05; **: P<0.01.</p

Expression of <i>TBX15</i>mRNA and <i>CXCL1</i>mRNA in BHT cell lines after stimulation with TNF-α and PMA/I.

<p>(A) qRT-PCR of <i>TBX15</i>mRNA analysis in BHT and BHT IkBαSR cells after 3h of TNF-α or PMA/I treatment. (B) qRT-PCR of <i>CXCL1</i>mRNA analysis in BHT and BHT IkBαSR cells after 3h of TNF-α or PMA/I treatment. Data were normalized to <i>RPL27</i> and expressed as fold change referred to untreated cells whose <i>TBX15</i>mRNA or <i>CXCL1</i>mRNA relative expression was defined as 1. Data are mean ± SD of mRNA levels in triplicates. ** indicates p-value < 0.01.</p

Identification of functional NF-κB responsive elements in the 5’-flanking region of the human <i>TBX15</i> gene.

<p>(A) Location and sequence of the predicted NF-κB binding sites (kBS1, kBS2 and kBS3) in the studied region of the <i>TBX15</i> promoter, with indication of altered sequence in mutated reporter constructs (underlined). Site directed mutagenesis was performed using P2 reporter construct to obtain the mutated reporter constructs. (B) Luciferase analysis of each mutated reporter constructs. <i>Left panel</i>, co-tansfected with the pCDNA3-p65 expression vector (p65) or the pCDNA3 empty vector (EV). <i>Right panel</i>, in nonstimulated or stimulated TNF-α cells. At least three independent experiments were performed with duplicates. Bars represent the mean±SD. * indicates p-value < 0.05 and ** p-value < 0.01.</p