571 results on '"Mauro Giacca"'
Search Results
52. Supplementary Figure 6 from Neuropilin-1 Identifies a Subset of Bone Marrow Gr1− Monocytes That Can Induce Tumor Vessel Normalization and Inhibit Tumor Growth
- Author
-
Mauro Giacca, Federico Bussolino, Enrico Giraudo, Guido Serini, Federica Maione, Milena Sinigaglia, Miguel Mano, Giulia Ruozi, Lorena Zentilin, Lucia Pattarini, Serena Zacchigna, Silvia Moimas, and Alessandro Carrer
- Abstract
PDF file - 1214K, Levels of expression of HIF-1 target genes in tumors inoculated with bone marrow Gr1+ cells or NEMs. Quantification of the mRNA levels of the indicated factors (VEGF-A, Angiopoietin-2 and Interleukin-6) was obtained by real-time PCR. The results are shown after normalization for the GAPDH gene. Data are mean �s.e.m. **=P
- Published
- 2023
53. Supplementary Figure 2 from Neuropilin-1 Identifies a Subset of Bone Marrow Gr1− Monocytes That Can Induce Tumor Vessel Normalization and Inhibit Tumor Growth
- Author
-
Mauro Giacca, Federico Bussolino, Enrico Giraudo, Guido Serini, Federica Maione, Milena Sinigaglia, Miguel Mano, Giulia Ruozi, Lorena Zentilin, Lucia Pattarini, Serena Zacchigna, Silvia Moimas, and Alessandro Carrer
- Abstract
PDF file - 3108K, Flow cytometry analysis of NEMs purified from the bone marrow. Flow cytometry analysis of bone marrow CD11b+/Gr1-/Nrp1+ cells (NEMs) using the indicated antibodies conjugated with phycoerythrin (PE). Percentages of positive cells, referred to the total number of NEMs, are reported. The plots reported in the first row are also shown in Figure 1D, fourth column.
- Published
- 2023
54. Supplementary Table 1 from Neuropilin-1 Identifies a Subset of Bone Marrow Gr1− Monocytes That Can Induce Tumor Vessel Normalization and Inhibit Tumor Growth
- Author
-
Mauro Giacca, Federico Bussolino, Enrico Giraudo, Guido Serini, Federica Maione, Milena Sinigaglia, Miguel Mano, Giulia Ruozi, Lorena Zentilin, Lucia Pattarini, Serena Zacchigna, Silvia Moimas, and Alessandro Carrer
- Abstract
PDF file - 91K, Supplementary Table 1. List of genes analyzed for expression by real-time, quantitative RT-PCR. All the amplifications were performed on a 7000 ABI Prism Instrument (Applied Biosystems), using pre-developed assays ("Mm" code), custom assays (for CCL5 and CXCR4) (Applied Biosystems) or SYBR Green (for Gr-1 and HPRT). The housekeeping genes GAPDH and HPRT were used to normalize the results.
- Published
- 2023
55. Supplementary Figure 1 from Neuropilin-1 Identifies a Subset of Bone Marrow Gr1− Monocytes That Can Induce Tumor Vessel Normalization and Inhibit Tumor Growth
- Author
-
Mauro Giacca, Federico Bussolino, Enrico Giraudo, Guido Serini, Federica Maione, Milena Sinigaglia, Miguel Mano, Giulia Ruozi, Lorena Zentilin, Lucia Pattarini, Serena Zacchigna, Silvia Moimas, and Alessandro Carrer
- Abstract
PDF file - 1560K, Quantification of the flow cytometry profiles for the characterization of the four bone marrow cell populations sorted according to the expression of Nrp1 and Gr1. The results (shown as bar charts in the upper part of the figure and as a table in the bottom part) refer to four independent experiments; shown are the mean � s.d. The markers analyzed are the same as in the representative experiment shown in Figure 1D.
- Published
- 2023
56. Supplementary Figure 4 from Neuropilin-1 Identifies a Subset of Bone Marrow Gr1− Monocytes That Can Induce Tumor Vessel Normalization and Inhibit Tumor Growth
- Author
-
Mauro Giacca, Federico Bussolino, Enrico Giraudo, Guido Serini, Federica Maione, Milena Sinigaglia, Miguel Mano, Giulia Ruozi, Lorena Zentilin, Lucia Pattarini, Serena Zacchigna, Silvia Moimas, and Alessandro Carrer
- Abstract
PDF file - 3482K, Proliferative activity of 4T1 tumors in vivo. A. Immunofluorescence staining of frozen tumor sections from mice injected with PBS, Gr1+ cells or NEMs using an antibody against PCNA (red); nuclei are counterstained in blue with DAPI. Analysis was performed after the animal sacrifice at day 18 after tumor inoculation. Scale bar: 100 �m. B. Quantitative analysis showing the percentage of PCNA+ nuclei in the tumor sections. Data are mean�s.e.m.; n=5 per group.
- Published
- 2023
57. Neutralization of NET-associated human ARG1 enhances cancer immunotherapy
- Author
-
Stefania Canè, Roza Maria Barouni, Marina Fabbi, John Cuozzo, Giulio Fracasso, Annalisa Adamo, Stefano Ugel, Rosalinda Trovato, Francesco De Sanctis, Mauro Giacca, Rita Lawlor, Aldo Scarpa, Borislav Rusev, Gabriella Lionetto, Salvatore Paiella, Roberto Salvia, Claudio Bassi, Susanna Mandruzzato, Silvano Ferrini, and Vincenzo Bronte
- Subjects
Pancreatic Neoplasms ,Mice ,Arginase ,Monoclonal ,Tumor Microenvironment ,Animals ,Humans ,Immunotherapy ,General Medicine ,CD8-Positive T-Lymphocytes ,Antibodies, Monoclonal ,Extracellular Traps ,Antibodies - Abstract
Myeloid cells can restrain antitumor immunity by metabolic pathways, such as the degradation of l -arginine, whose concentrations are regulated by the arginase 1 (ARG1) enzyme. Results from preclinical studies indicate the important role of arginine metabolism in pancreatic ductal adenocarcinoma (PDAC) progression, suggesting a potential for clinical application; however, divergent evolution in ARG1 expression and function in rodents and humans has restricted clinical translation. To overcome this dichotomy, here, we show that neutrophil extracellular traps (NETs), released by spontaneously activated neutrophils isolated from patients with PDAC, create a microdomain where cathepsin S (CTSS) cleaves human (h)ARG1 into different molecular forms endowed with enhanced enzymatic activity at physiological pH. NET-associated hARG1 suppresses T lymphocytes whose proliferation is restored by either adding a hARG1-specific monoclonal antibody (mAb) or preventing CTSS-mediated cleavage, whereas small-molecule inhibitors are not effective. We show that ARG1 blockade, combined with immune checkpoint inhibitors, can restore CD8 + T cell function in ex vivo PDAC tumors. Furthermore, anti-hARG1 mAbs increase the frequency of adoptively transferred tumor-specific CD8 + T cells in tumor and enhance the effectiveness of immune checkpoint therapy in humanized mice. Thus, this study shows that extracellular ARG1, released by activated myeloid cells, localizes in NETs, where it interacts with CTSS that in turn cleaves ARG1, producing major molecular forms endowed with different enzymatic activity at physiological pH. Once exocytosed, ARG1 activity can be targeted by mAbs, which bear potential for clinical application for the treatment of PDAC and require further exploration.
- Published
- 2023
58. The pathological maelstrom of COVID-19 and cardiovascular disease
- Author
-
Mauro Giacca and Ajay M. Shah
- Published
- 2022
59. 525 PROGNOSTIC RELEVANCE OF DEMOGRAPHIC FACTORS IN CARDIAC MAGNETIC RESONANCE-PROVEN ACUTE MYOCARDITIS. A COHORT STUDY
- Author
-
Antonio Cannata, Roman Roy, Antonio Jordan Rios, Emma Ferone, Barbara Cassimon, Matthew Sadler, Irfan Rind, Stefania Rosmini, Susan Piper, Mauro Giacca, Theresa Mcdonagh, Paul Scott, and Daniel Bromage
- Subjects
Cardiology and Cardiovascular Medicine - Abstract
Aims Acute myocarditis (AM) is a heterogeneous condition with variable estimates of survival. Contemporary criteria for the diagnosis of clinically suspected AM enable non-invasive assessment, resulting in greater sensitivity and more representative cohorts. We aimed to describe the demographic characteristics and long-term outcomes of patients with AM diagnosed using non-invasive criteria. Methods and Results A total of 199 patients with cardiac magnetic resonance (CMR)-confirmed AM were included. The majority (n=130, 65%) were male and the average age was 39±16 years. Half of patients were White (n=99, 52%) with the remainder from Black and Minority Ethnic (BAME) groups. The most common clinical presentation was with chest pain (n=156, 78%), with smaller numbers presenting with breathlessness (n=25, 13%) and arrhythmias (n=18, 9%). Patients admitted with breathlessness were sicker and more often required inotropes, steroids, and renal replacement therapy (p Conclusions AM is highly heterogeneous with an overall favourable prognosis. Three quarters of patients with AM present with chest pain and this is associated with a benign prognosis. AM presenting with life-threatening arrhythmias is associated with higher risk of adverse events.
- Published
- 2022
60. Tumor vascular remodeling by thrombospondin-1 enhances drug delivery and antineoplastic activity
- Author
-
Andrea Resovi, Patrizia Borsotti, Fabio Sangalli, Enrico Davoli, Alexander Berndt, Lorena Zentilin, Massimo Zucchetti, Raffaella Giavazzi, Alice Passoni, Dorina Belotti, Giovanni Valbusa, Denise Pinessi, Mauro Giacca, Giulia Taraboletti, Lavinia Morosi, and Elena Carlessi
- Subjects
Cisplatin ,Tumor microenvironment ,Neovascularization, Pathologic ,business.industry ,Angiogenesis ,Angiogenesis Inhibitors ,Antineoplastic Agents ,Vascular Remodeling ,chemistry.chemical_compound ,Pharmaceutical Preparations ,Paclitaxel ,chemistry ,In vivo ,Drug delivery ,Thrombospondin 1 ,Cancer research ,Humans ,Medicine ,Ectopic expression ,business ,Molecular Biology ,medicine.drug - Abstract
The disorganized and inefficient tumor vasculature is a major obstacle to the delivery and efficacy of antineoplastic treatments. Antiangiogenic agents can normalize the tumor vessels, improving vessel function and boosting the distribution and activity of chemotherapy. The type III repeats (T3R) domain of thrombospondin-1 contains different potential antiangiogenic sequences. We therefore hypothesized that it might affect the tumor vasculature. Ectopic expression of the T3R domain by the tumor cells or by the host, or administration of recombinant T3R, delayed the in vivo growth of experimental tumors. Tumors presented marked reorganization of the vasculature, with abundant but smaller vessels, associated with substantially less necrosis. Mechanistically, the use of truncated forms of the domain, containing different active sequences, pointed to the FGF2/FGFR/ERK axis as a target for T3R activity. Along with reduced necrosis, the expression of T3R promoted tumor distribution of chemotherapy (paclitaxel), with a higher drug concentration and more homogeneous distribution, as assessed by HPLC and MALDI imaging mass spectrometry. T3R-expressing tumors were more responsive to paclitaxel and cisplatin. This study shows that together with its known role as a canonical inhibitor of angiogenesis, thrombospondin-1 can also remodel tumor blood vessels, affecting the morphological and functional properties of the tumor vasculature. The ability of T3R to reduce tumor growth and improve the response to chemotherapy opens new perspectives for therapeutic strategies based on T3R to be used in combination therapies.
- Published
- 2021
61. SARS-CoV-2, myocardial injury and inflammation: insights from a large clinical and autopsy study
- Author
-
Marco Metra, Rossana Bussani, Alessia Paldino, Fulvio Silvestri, Serena Zacchigna, Chiara Collesi, Mauro Giacca, Marco Merlo, Matteo Dal Ferro, Ricardo Correa, Gianfranco Sinagra, Edoardo Schneider, Lorena Zentilin, Vincenzo Nuzzi, Dal Ferro, M., Bussani, R., Paldino, A., Nuzzi, V., Collesi, C., Zentilin, L., Schneider, E., Correa, R., Silvestri, F., Zacchigna, S., Giacca, M., Metra, M., Merlo, M., and Sinagra, G.
- Subjects
Male ,0301 basic medicine ,Pathology ,Necrosis ,Myocarditi ,Autopsy ,Disease ,030204 cardiovascular system & hematology ,Severity of Illness Index ,Cohort Studies ,0302 clinical medicine ,80 and over ,Myocytes, Cardiac ,Aged, 80 and over ,education.field_of_study ,General Medicine ,Myocarditis ,Cardiac autopsy study ,COVID-19 ,Myocardial injury ,Cohort ,Cardiology ,Immunohistochemistry ,Female ,medicine.symptom ,Cardiology and Cardiovascular Medicine ,Cardiac ,medicine.medical_specialty ,Population ,Inflammation ,03 medical and health sciences ,Internal medicine ,medicine ,Humans ,education ,Aged ,Original Paper ,Myocytes ,SARS-CoV-2 ,business.industry ,Myocardium ,Correction ,medicine.disease ,030104 developmental biology ,business - Abstract
Objective Despite growing evidence about myocardial injury in hospitalized COronaVIrus Disease 2019 (COVID-19) patients, the mechanism behind this injury is only poorly understood and little is known about its association with SARS-CoV-2-mediated myocarditis. Furthermore, definite evidence of the presence and role of SARS-CoV-2 in cardiomyocytes in the clinical scenario is still lacking. Methods We histologically characterized myocardial tissue of 40 patients deceased with severe SARS-CoV-2 infection during the first wave of the pandemic. Clinical data were also recorded and analyzed. In case of findings supportive of myocardial inflammation, histological analysis was complemented by RT-PCR and immunohistochemistry for SARS-CoV-2 viral antigens and in situ RNA hybridization for the detection of viral genomes. Results Both chronic and acute myocardial damage was invariably present, correlating with the age and comorbidities of our population. Myocarditis of overt entity was found in one case (2.5%). SARS-CoV-2 genome was not found in the cardiomyocytes of the patient with myocarditis, while it was focally and negligibly present in cardiomyocytes of patients with known viral persistence in the lungs and no signs of myocardial inflammation. The presence of myocardial injury was not associated with myocardial inflammatory infiltrates. Conclusions In this autopsy cohort of COVID-19 patients, myocarditis is rarely found and not associated with SARS-CoV-2 presence in cardiomyocytes. Chronic and acute forms of myocardial damage are constantly found and correlate with the severity of COVID-19 disease and pre-existing comorbidities. Graphic abstract
- Published
- 2021
62. Abstract P1019: CycleTrack , A Genetic Method To Trace Cardiomyocyte Renewal In Small And Large Animal Models
- Author
-
Ilaria Secco, Ana Backovic, Simone Vodret, Matteo Dal Ferro, Mateusz Tomczyk, Khatia Gabisonia, Lucia Carlucci, Lorena Zentilin, Serena Zacchigna, Fabio A Recchia, and Mauro Giacca
- Subjects
Physiology ,Cardiology and Cardiovascular Medicine - Abstract
To what extent cardiac renewal occurs in adulthood and can be stimulated by therapeutic interventions is still a matter of investigation. In particular, over the last few years, significant effort has been made into awakening the endogenous regenerative potential of the adult mammalian heart after injury using different strategies. However, these studies are significantly hindered by the lack of tools to properly assess cardiomyocyte renewal over time. We have developed a genetic strategy, which we named CycleTrack, allowing a cumulative and accurate estimate of cardiomyocyte divisions in vivo. This method is based on the expression, in cardiomyocytes, of the Cre recombinase under the control of a 312-bp fragment of the Cyclin B2 promoter, which is specifically sensitive to cell proliferation. Replication-activated Cre acts on floxed GFP that is either present in the genome of transgenic mice or is exogenously delivered using Adeno-Associated Viral (AAV) vectors. As a result, cardiomyocytes traversing G2/M become irreversibly labeled. Using CycleTrack, we could monitor cardiomyocyte turnover in several physiological and pathological conditions. These included the measurement of the rate of proliferation of cardiomyocytes after apical resection in neonatal mice, the pro-regenerative effect of AAV9-mediated delivery of miRNA-199a and miRNA-590 after myocardial infarction in adult mice and the effect of pregnancy on myocardial hyperplasia. We also delivered the CycleTrack vectors into pig hearts to visualize cardiomyocyte turnover after myocardial infarction. Considering the large availability of Cre reporter mouse lines and the efficacy of AAVs to transfer genes into cardiomyocytes, we propose CycleTrack as a robust and straightforward tool to trace mitotic events for cardiac regeneration studies in small and large animal models.
- Published
- 2022
63. Abstract P1002: Elucidating The Role Of Dyrk1a Kinase In Controlling Cardiomyocyte Proliferation
- Author
-
Giorgia Rizzari, Ellen Dirkx, Consuelo Torrini, Mateusz Tomczyk, Miguel Mano, and Mauro Giacca
- Subjects
Physiology ,Cardiology and Cardiovascular Medicine - Abstract
Despite the development of new pharmacological and device-based therapies, no approaches currently exist to compensate for the irreversible loss of cardiomyocytes occurring after myocardial infarction. In a search for novel treatments that induce cardiomyocyte proliferation, we performed a high-content, fluorescence-microscopy-based, high-throughput screening in neonatal rat cardiomyocytes, using a library of FDA/EMA-approved drug library compounds. The screening, which was based on EdU incorporation as an S-phase marker, identified harmine as the most powerful drug enhancing proliferation in combination with miR-199a-3p and miR-590-3p, the most effective microRNAs that the laboratory had previously identified. Harmine treatment of CD1 mice showed an improved cardiac outcome upon harmine treatment compared to controls. Harmine is a potent inhibitor of the Dyrk1 family of protein kinases, with high specificity for the Dyrk1a isoform. Dyrk1a downregulation experiments in neonatal rat ventricular cardiomyocytes showed increased proliferation, consistent with a negative role of this kinase. The simultaneous knockdown of YAP1 blunted the effect of Dyrk1a downregulation. Of note, the YAP total protein levels were unchanged upon either Dyrk1a silencing or overexpression, but there was a marked modification of YAP subcellular localisation, being YAP1 nuclear upon knockdown and mostly cytoplasmic upon overexpression. Dyrk1a downregulation and harmine treatment activated YAP-controlled gene expression. In the course of these experiments, we also noticed a reciprocal regulation of YAP onto DYRK1a, as YAP downregulation markedly decreased Dyrk1a protein levels. This finding was reinforced by the observations that, in cardiomyocytes, the YAP-TEAD inhibitor verteporfin also decreased Dyrk1a protein levels. Of note, this downregulation was not rescued by the proteasome inhibitor MG132, thus indicating that the level of regulation is most likely transcriptional. These results support the role of Dyrk1a as an anti-proliferative factor in cardiomyocytes and highlight a novel feedback connection between Dyrk1a and YAP.
- Published
- 2022
64. CRISPR activation and interference as investigative tools in the cardiovascular system
- Author
-
Melissa S. Carroll and Mauro Giacca
- Subjects
Cell Biology ,Biochemistry - Abstract
CRISPR activation and interference (CRISPRa/i) technology offers the unprecedented possibility of achieving regulated gene expression both in vitro and in vivo. The DNA pairing specificity of a nuclease dead Cas9 (dCas9) is exploited to precisely target a transcriptional activator or repressor in proximity to a gene promoter. This permits both the study of phenotypes arising from gene modulation for investigative purposes, and the development of potential therapeutics. As with virtually all other organ systems, the cardiovascular system can deeply benefit from a broader utilisation of CRISPRa/i. However, application of this technology is still in its infancy. Significant areas for improvement include the identification of novel and more effective transcriptional regulators that can be docked to dCas9, and the development of more efficient methods for their delivery and expression in vivo.
- Published
- 2022
65. CARMN Loss Regulates Smooth Muscle Cells and Accelerates Atherosclerosis in Mice
- Author
-
Marion J.J. Gijbels, Mukesh Kumar Lalwani, Matthew R. Bennett, Jessica P. Scanlon, Dónal O'Carroll, Kim van Kuijk, Francesca Vacante, David E. Newby, Amanda C Doran, Amira D. Mahmoud, Andrew H. Baker, Thomas C Williams, Judith C. Sluimer, Patrick W. F. Hadoke, Mauro Giacca, Michael B. Clark, Laura Denby, Rusty L. Montgomery, Jenny de Bruijn, Eileen Miller, Julie Rodor, Azzurra L De Pace, Medical Biochemistry, ACS - Atherosclerosis & ischemic syndromes, Pathologie, and RS: Carim - B07 The vulnerable plaque: makers and markers
- Subjects
Vascular smooth muscle ,Physiology ,phenotype ,LOW-DENSITY-LIPOPROTEIN ,Inflammation ,PROGRESSION ,Biology ,Article ,MIR-145 ,Transcriptome ,ACTIVATION ,In vivo ,PHENOTYPIC MODULATION ,microRNA ,medicine ,COLLAGENS ,Humans ,Cell growth ,LONG NONCODING RNA ,PROLIFERATION ,cholesterol ,Phenotype ,Long non-coding RNA ,EVOLUTION ,Cell biology ,DIFFERENTIATION ,cell proliferation ,inflammation ,RNA, Long Noncoding ,medicine.symptom ,atherosclerosis ,Cardiology and Cardiovascular Medicine - Abstract
Rationale: In the microenvironment of atherosclerotic lesions, vascular smooth muscle cells (vSMCs) switch to a dedifferentiated state but the underlying molecular mechanisms driving this switch are not fully understood. Long noncoding RNAs (lncRNAs) are dysregulated during vascular pathology, but relatively little is known about their involvement in controlling vSMCs function. Cardiac mesoderm enhancer-associated noncoding RNA (CARMN) is a lncRNA located immediately upstream of the microRNAs-143 and -145 (miR-143 and miR-145 ), both involved in vSMCs function. Objective: We investigated the role of the lncRNA CARMN, independent from miR-143 and miR-145, as a potential regulator of vSMC phenotypes in vitro and the consequences of its loss during the development of atherosclerosis in vivo. We hypothesized that loss of CARMN is a primary event controlling the functional switch towards proatherogenic vSMC phenotype and accelerates the development of the plaques in vivo. Method and Results: Expression of CARMN lncRNA was silenced using locked nucleic acids antisense oligonucleotides (GapmeRs) in human coronary arterial smooth muscle cells, revealing that GapmeR-mediated loss of CARMN negatively affects miR-143 and miR-145 microRNA expression. RNA sequencing of CARMN-depleted human coronary arterial smooth muscle cells revealed large transcriptomic changes, associated with vSMC proliferation, migration, inflammation, lipid metabolism, and dedifferentiation. The use of miR-143 and miR-145 mimics revealed that CARMN regulates human coronary arterial smooth muscle cell proliferation in a microRNA-independent manner. In humans and mice, CARMN and associated microRNAs were downregulated in advanced versus early atherosclerotic lesions. Using a CRISPR (clustered regularly interspaced short palindromic repeats)-Cas9 (CRISPR-associated protein 9) knockout approach, we explored the implications of CARMN depletion during atherosclerosis in vivo. Consistent with in vitro results, the knockout of CARMN impaired the expression of miR-143 and miR-145 under homeostatic conditions. Importantly, when atherosclerosis was induced in these mice, CARMN knockout increased the volume, size, proinflammatory Lgals3 (galectin 3)-expressing cells content, and altered plaque composition, yielding an advanced phenotype. Conclusions: We identified the early loss of CARMN lncRNA as critical event which primes vSMCs towards a proatherogenic phenotype in vitro and accelerates the development of atherosclerosis in vivo.
- Published
- 2021
66. Is heart failure with preserved ejection fraction a ‘dementia’ of the heart?
- Author
-
Mauro Giacca, Matteo Pardini, Gianfranco Sinagra, Marco Canepa, Niccolò Marchionni, Iacopo Olivotto, Federica del Monte, Giacomo Tini, Antonio Cannatà, Pier Giorgio Masci, James E. Udelson, Tini, G., Cannata, A., Canepa, M., Masci, P. G., Pardini, M., Giacca, M., Sinagra, G., Marchionni, N., Del Monte, F., Udelson, J. E., and Olivotto, I.
- Subjects
Cardiac function curve ,medicine.medical_specialty ,Diastole ,030204 cardiovascular system & hematology ,Left ventricular hypertrophy ,Ventricular Function, Left ,03 medical and health sciences ,0302 clinical medicine ,Myocardial fibrosis ,Internal medicine ,Humans ,Medicine ,Dementia ,Calcium handling ,030212 general & internal medicine ,Cardiac aging ,HFpEF ,Pathological ,Heart Failure ,business.industry ,Heart ,Stroke Volume ,medicine.disease ,Physiological Aging ,Heart failure ,Cardiology ,Cardiology and Cardiovascular Medicine ,Heart failure with preserved ejection fraction ,business - Abstract
Heart failure with preserved ejection fraction (HFpEF) remains an elusive entity, due to its heterogeneous clinical profile and an arbitrarily defined nosology. Several pathophysiological mechanisms recognized as central for the development of HFpEF appear to be in common with the process of physiological aging of the heart. Both conditions are characterized by progressive impairment in cardiac function, accompanied by left ventricular hypertrophy, diastolic dysfunction, sarcomeric, and metabolic abnormalities. The neurological paradigm of dementia-intended as a progressive, multifactorial organ damage with decline of functional reserve, eventually leading to irreversible dysfunction-is well suited to represent HFpEF. In such perspective, certain phenotypes of HFpEF may be viewed as a maladaptive response to environmental modifiers, causing premature and pathological aging of the heart. We here propose that the 'HFpEF syndrome' may reflect the interplay of adverse structural remodelling and erosion of functional reserve, mirroring the processes leading to dementia in the brain. The resulting conceptual framework may help advance our understanding of HFpEF and unravel potential therapeutical targets.
- Published
- 2021
67. Molecular tools to detect the activation of key signalling pathways driving cardiomyocyte proliferation
- Author
-
Rebecca Artioli, Karim Rahhali, Lorena Zentilin, Chiara Collesi, and Mauro Giacca
- Subjects
Cardiology and Cardiovascular Medicine ,Molecular Biology - Published
- 2022
68. Cardioprotective effect of Fam3c after ischemic damage
- Author
-
Antonio Mura, Francesca Bortolotti, Giulia Ruozi, Mateusz Tomczyk, Edoardo Schneider, Simone Vodret, Giulio Ciucci, Lorena Zentilin, and Mauro Giacca
- Subjects
Cardiology and Cardiovascular Medicine ,Molecular Biology - Published
- 2022
69. Gene editing for cardiomyopathy takes a step forward
- Author
-
MAURO GIACCA
- Subjects
Physiology ,Physiology (medical) ,Cardiology and Cardiovascular Medicine - Published
- 2022
70. Prognostic Prediction of Genotype vs Phenotype in Genetic Cardiomyopathies
- Author
-
Alessia Paldino, Matteo Dal Ferro, Davide Stolfo, Ilaria Gandin, Kristen Medo, Sharon Graw, Marta Gigli, Giulia Gagno, Denise Zaffalon, Matteo Castrichini, Marco Masè, Antonio Cannatà, Francesca Brun, Garrett Storm, Giovanni Maria Severini, Stefania Lenarduzzi, Giorgia Girotto, Paolo Gasparini, Francesca Bortolotti, Mauro Giacca, Serena Zacchigna, Marco Merlo, Matthew R.G. Taylor, Luisa Mestroni, Gianfranco Sinagra, Paldino, Alessia, Dal Ferro, Matteo, Stolfo, Davide, Gandin, Ilaria, Medo, Kristen, Graw, Sharon, Gigli, Marta, Gagno, Giulia, Zaffalon, Denise, Castrichini, Matteo, Masè, Marco, Cannatà, Antonio, Brun, Francesca, Storm, Garrett, Severini, Giovanni Maria, Lenarduzzi, Stefania, Girotto, Giorgia, Gasparini, Paolo, Bortolotti, Francesca, Giacca, Mauro, Zacchigna, Serena, Merlo, Marco, Taylor, Matthew R G, Mestroni, Luisa, and Sinagra, Gianfranco
- Subjects
Cardiomyopathy, Dilated ,DCM ,Genotype ,phenotype ,genotype ,Arrhythmias, Cardiac ,pathogenic/likely pathogenic variants ,Prognosis ,pathogenic/likely pathogenic variant ,ALVC ,Phenotype ,Death, Sudden, Cardiac ,ARVC ,Cytidine Monophosphate ,Humans ,Cardiology and Cardiovascular Medicine ,Cardiomyopathies - Abstract
Background: Diverse genetic backgrounds often lead to phenotypic heterogeneity in cardiomyopathies (CMPs). Previous genotype-phenotype studies have primarily focused on the analysis of a single phenotype, and the diagnostic and prognostic features of the CMP genotype across different phenotypic expressions remain poorly understood. Objectives: We sought to define differences in outcome prediction when stratifying patients based on phenotype at presentation compared with genotype in a large cohort of patients with CMPs and positive genetic testing. Methods: Dilated cardiomyopathy (DCM), arrhythmogenic right ventricular cardiomyopathy, left-dominant arrhythmogenic cardiomyopathy, and biventricular arrhythmogenic cardiomyopathy were examined in this study. A total of 281 patients (80% DCM) with pathogenic or likely pathogenic variants were included. The primary and secondary outcomes were: 1) all-cause mortality (D)/heart transplant (HT); 2) sudden cardiac death/major ventricular arrhythmias (SCD/MVA); and 3) heart failure-related death (DHF)/HT/left ventricular assist device implantation (LVAD). Results: Survival analysis revealed that SCD/MVA events occurred more frequently in patients without a DCM phenotype and in carriers of DSP, PKP2, LMNA, and FLNC variants. However, after adjustment for age and sex, genotype-based classification, but not phenotype-based classification, was predictive of SCD/MVA. LMNA showed the worst trends in terms of D/HT and DHF/HT/LVAD. Conclusions: Genotypes were associated with significant phenotypic heterogeneity in genetic cardiomyopathies. Nevertheless, in our study, genotypic-based classification showed higher precision in predicting the outcome of patients with CMP than phenotype-based classification. These findings add to our current understanding of inherited CMPs and contribute to the risk stratification of patients with positive genetic testing.
- Published
- 2022
71. Regenerative potential of epicardium-derived extracellular vesicles mediated by conserved miRNA transfer
- Author
-
Moritz Matthaei, Tahnee Kennedy, Mala Gunadasa-Rohling, Cristina Villa del Campo, Paul R. Riley, Norman Y. Liaw, Luca Braga, Wolfram-Hubertus Zimmermann, Mauro Giacca, Niels Voigt, and Gabriela Salinas
- Subjects
Time Factors ,Physiology ,medicine.medical_treatment ,Human Embryonic Stem Cells ,Myocardial Infarction ,030204 cardiovascular system & hematology ,Regenerative Medicine ,Coronary artery disease ,0302 clinical medicine ,Medicine ,Myocytes, Cardiac ,AcademicSubjects/MED00200 ,Myocardial infarction ,Zebrafish ,Heart transplantation ,FUCCI ,Human engineered myocardium ,0303 health sciences ,education.field_of_study ,biology ,MicroRNA ,Epicardium ,Cell biology ,Cardiology and Cardiovascular Medicine ,Pericardium ,Population ,Exosome ,Cell Line ,Extracellular Vesicles ,03 medical and health sciences ,Physiology (medical) ,Paracrine Communication ,Animals ,Humans ,Regeneration ,education ,Cell Proliferation ,030304 developmental biology ,business.industry ,Regeneration (biology) ,Recovery of Function ,Original Articles ,medicine.disease ,biology.organism_classification ,Myocardial Contraction ,Rats ,Mice, Inbred C57BL ,Disease Models, Animal ,MicroRNAs ,Animals, Newborn ,Heart failure ,business - Abstract
Aims After a myocardial infarction, the adult human heart lacks sufficient regenerative capacity to restore lost tissue, leading to heart failure progression. Finding novel ways to reprogram adult cardiomyocytes into a regenerative state is a major therapeutic goal. The epicardium, the outermost layer of the heart, contributes cardiovascular cell types to the forming heart and is a source of trophic signals to promote heart muscle growth during embryonic development. The epicardium is also essential for heart regeneration in zebrafish and neonatal mice and can be reactivated after injury in adult hearts to improve outcome. A recently identified mechanism of cell–cell communication and signalling is that mediated by extracellular vesicles (EVs). Here, we aimed to investigate epicardial signalling via EV release in response to cardiac injury and as a means to optimize cardiac repair and regeneration. Methods and results We isolated epicardial EVs from mouse and human sources and targeted the cardiomyocyte population. Epicardial EVs enhanced proliferation in H9C2 cells and in primary neonatal murine cardiomyocytes in vitro and promoted cell cycle re-entry when injected into the injured area of infarcted neonatal hearts. These EVs also enhanced regeneration in cryoinjured engineered human myocardium (EHM) as a novel model of human myocardial injury. Deep RNA-sequencing of epicardial EV cargo revealed conserved microRNAs (miRs) between human and mouse epicardial-derived exosomes, and the effects on cell cycle re-entry were recapitulated by administration of cargo miR-30a, miR-100, miR-27a, and miR-30e to human stem cell-derived cardiomyocytes and cryoinjured EHM constructs. Conclusion Here, we describe the first characterization of epicardial EV secretion, which can signal to promote proliferation of cardiomyocytes in infarcted mouse hearts and in a human model of myocardial injury, resulting in enhanced contractile function. Analysis of exosome cargo in mouse and human identified conserved pro-regenerative miRs, which in combination recapitulated the therapeutic effects of promoting cardiomyocyte proliferation., Graphical Abstract
- Published
- 2021
72. Characterization of Viral Genome Encapsidated in Adeno-associated Recombinant Vectors Produced in Yeast Saccharomyces cerevisiae
- Author
-
Veronica Della Latta, Ilenia Iaia, Mauro Giacca, Lorena Zentilin, Tiziana Cervelli, Filippo Cipriani, Alvaro Galli, and Maria Serena Milella
- Subjects
0106 biological sciences ,viruses ,Saccharomyces cerevisiae ,Bioengineering ,Biology ,medicine.disease_cause ,01 natural sciences ,Applied Microbiology and Biotechnology ,Biochemistry ,Genome ,law.invention ,03 medical and health sciences ,law ,010608 biotechnology ,medicine ,Vector (molecular biology) ,Molecular Biology ,Adeno-associated virus ,Gene ,030304 developmental biology ,0303 health sciences ,biology.organism_classification ,Virology ,Yeast ,Capsid ,Recombinant DNA ,Biotechnology - Abstract
Adeno-associated virus (AAV) is a small, non-enveloped virus used as vector in gene therapy, mainly produced in human cells and in baculovirus systems. Intense studies on these platforms led to the production of vectors with titers between 103 and 105 viral genomes (vg) per cells. In spite of this, vector yields need to be improved to satisfy the high product demands of clinical trials and future commercialization. Our studies and those of other groups have explored the possibility to exploit the yeast Saccharomyces cerevisiae to produce rAAV. We previously demonstrated that yeast supports AAV genome replication and capsid assembly. The purpose of this study was to evaluate the quality of the encapsidated AAV DNA. Here, we report the construction of a yeast strain expressing Rep68/40 from an integrated copy of the Rep gene under the control of the yeast constitutive ADH promoter and Capsid proteins from the Cap gene under the control of an inducible GAL promoter. Our results indicate that a portion of AAV particles generated by this system contains encapsidated AAV DNA. However, the majority of encapsidated DNA consists of fragmented regions of the transgene cassette, with ITRs being the most represented sequences. Altogether, these data indicate that, in yeast, encapsidation occurs with low efficiency and that rAAVs resemble pseudo-vectors that are present in clinical-grade rAAV preparations.
- Published
- 2021
73. Nrf2 attenuates the innate immune response after experimental myocardial infarction
- Author
-
Daniel I. Bromage, Silvia C. Trevelin, Josef Huntington, Victoria X. Yang, Ananya Muthukumar, Sarah J. Mackie, Greta Sawyer, Xiaohong Zhang, Celio X.C. Santos, Niloufar Safinia, Ioannis Smyrnias, Mauro Giacca, Aleksandar Ivetic, and Ajay M. Shah
- Subjects
Mice, Knockout ,Ventricular Remodeling ,NF-E2-Related Factor 2 ,Myocardium ,Biophysics ,Myocardial Infarction ,Cell Biology ,Biochemistry ,Immunity, Innate ,Mice, Inbred C57BL ,Disease Models, Animal ,Mice ,Animals ,Cytokines ,Molecular Biology - Abstract
ObjectivesWe aimed to investigate the contribution of the transcription factor nuclear factor erythroid-derived 2-like 2 (Nrf2) to the inflammatory response after experimental myocardial infarction (MI).BackgroundThere is compelling evidence implicating dysregulated inflammation in the mechanism of ventricular remodeling and heart failure (HF) after MI. The transcription factor Nrf2 (encoded by Nfe2l2) is a promising target in this context. It impedes transcriptional upregulation of pro-inflammatory cytokines and is anti-inflammatory in various murine models.MethodsWe subjected Nrf2-/- mice and wild type (WT) controls to permanent left coronary artery (LCA) ligation. The inflammatory response was investigated with fluorescence-activated cell sorting (FACS) analysis of peripheral blood and heart cell suspensions, together with qRT-PCR of infarcted tissue for chemokines and their receptors. To investigate whether Nrf2-mediated transcription is a dedicated function of leukocytes, we interrogated publicly available RNA-sequencing (RNA-seq) data from mouse hearts after permanent LCA ligation for Nrf2-regulated gene (NRG) expression.ResultsFACS analysis demonstrated a profoundly inflamed phenotype in the hearts of global Nrf2-/- mice as compared to WT mice after MI. Moreover, infarcted tissue from Nrf2-/- mice displayed higher expression of inflammatory cytokines, chemokines, and their receptors, including IL6, Ccl2, and Cxcr4. RNA-seq analysis showed upregulated NRG expression in WT mice after MI compared to untreated mice, which was significantly higher in bioinformatically isolated CCR2+ cells.ConclusionsTaken together, the results suggest that Nrf2 signalling in leukocytes, and possibly CCR2+ monocyte-derived cardiac resident macrophages, may be potential targets to prevent post-MI ventricular remodeling.
- Published
- 2022
74. n-3 PUFA dietary lipid replacement normalizes muscle mitochondrial function and oxidative stress through enhanced tissue mitophagy and protects from muscle wasting in experimental kidney disease
- Author
-
Gianluca, Gortan Cappellari, Annamaria, Semolic, Giulia, Ruozi, Davide, Barbetta, Francesca, Bortolotti, Pierandrea, Vinci, Michela, Zanetti, Robert H, Mak, Giacomo, Garibotto, Mauro, Giacca, Rocco, Barazzoni, Gortan Cappellari, Gianluca, Semolic, Annamaria, Ruozi, Giulia, Barbetta, Davide, Bortolotti, Francesca, Vinci, Pierandrea, Zanetti, Michela, Mak, Robert H, Garibotto, Giacomo, Giacca, Mauro, and Barazzoni, Rocco
- Subjects
Endocrinology, Diabetes and Metabolism ,Mitophagy ,Skeletal muscle ,Dietary Fats ,Rats ,Mitochondria ,CKD ,Uremia ,n3-PUFA ,Muscular Atrophy ,Oxidative Stress ,Adenosine Triphosphate ,Endocrinology ,Fatty Acids, Omega-3 ,Animals ,Renal Insufficiency, Chronic ,Muscle, Skeletal ,Reactive Oxygen Species - Abstract
Introduction and methods: Skeletal muscle mitochondrial dysfunction may cause tissue oxidative stress and consequent catabolism in chronic kidney disease (CKD), contributing to patient mortality. We investigated in 5/ 6-nephrectomized (Nx) rats the impact of n3-polyunsaturated fatty-acids (n3-PUFA) isocaloric partial dietary replacement on gastrocnemius muscle (Gm) mitochondrial master-regulators, ATP production, ROS generation and related muscle-catabolic derangements. Results: Nx had low Gm mitochondrial nuclear respiratory factor-2 and peroxisome proliferator-activated receptor gamma coactivator-1alpha, low ATP production and higher mitochondrial fission-fusion protein ratio with ROS overproduction. n3-PUFA normalized all mitochondrial derangements and pro-oxidative tissue redox state (oxydized to total glutathione ratio). n3-PUFA also normalized Nx-induced muscle-catabolic proinflammatory cytokines, insulin resistance and low muscle weight. Human uremic serum reproduced mitochondrial derangements in C2C12 myotubes, while n3-PUFA coincubation prevented all effects. n3-PUFA also enhanced muscle mitophagy in-vivo and siRNA-mediated autophagy inhibition selectively blocked n3-PUFAinduced normalization of C2C12 mitochondrial ROS production. Conclusions: In conclusion, dietary n3-PUFA normalize mitochondrial master-regulators, ATP production and dynamics in experimental CKD. These effects occur directly in muscle cells and they normalize ROS production through enhanced mitophagy. Dietary n3-PUFA mitochondrial effects result in normalized catabolic derangements and protection from muscle wasting, with potential positive impact on patient survival.
- Published
- 2022
75. Cardiovascular Damage in COVID-19: What We Know Two Years Later
- Author
-
Vincenzo Nuzzi, Eva Del Mestre, Alessia Degrassi, Daniel I. Bromage, Paolo Manca, Susan Piper, Jessica Artico, Piero Gentile, Paul A. Scott, Mario Chiatto, Marco Merlo, Nilesh Pareek, Mauro Giacca, Gianfranco Sinagra, Theresa A. McDonagh, Antonio Cannata, Nuzzi, Vincenzo, Del Mestre, Eva, Degrassi, Alessia, Bromage, Daniel I, Manca, Paolo, Piper, Susan, Artico, Jessica, Gentile, Piero, Scott, Paul A, Chiatto, Mario, Merlo, Marco, Pareek, Nilesh, Giacca, Mauro, Sinagra, Gianfranco, Mcdonagh, Theresa A, and Cannata, Antonio
- Subjects
Acute cardiac damage ,Heart Diseases ,SARS-CoV-2 ,Humans ,COVID-19 ,Heart ,Cardiovascular injury ,Biomarker ,Cardiology and Cardiovascular Medicine ,Pandemics ,Biomarkers - Abstract
Purpose of the Review The Coronavirus disease 2019 (COVID-19) pandemic has profoundly influenced cardiological clinical and basic research in the past two years. In the present review, we summarize the current knowledge on myocardial involvement in COVID-19, providing an overview on the incidence, the pathogenetic mechanisms, and the clinical implications of cardiac injury in this setting. Recent Findings The possibility of heart involvement in patients with COVID-19 has received great attention since the beginning of the pandemic. After more than two years, several steps have been taken in understanding the mechanisms and the incidence of cardiac injury during COVID-19 infection. Similarly, studies globally have clarified the implications of co-existing heart disease and COVID-19. Summary Severe COVID-19 infection may be complicated by myocardial injury. To date, a direct damage from the virus has not been demonstrated. The presence of myocardial injury should be systematically assessed for a prognostication purpose and for possible therapeutic implications.
- Published
- 2022
76. Scientists on the Spot: Re-awakening the heart’s regenerative capacity
- Author
-
Monika M Gladka, Mauro Giacca, Medical Biology, and ACS - Heart failure & arrhythmias
- Subjects
Endogenous ,Physiology ,Regeneration (biology) ,MicroRNA ,Biology ,Embryonic stem cell ,Cell biology ,Cardiac repair ,Physiology (medical) ,microRNA ,Regeneration ,Embryonic ,Cardiology and Cardiovascular Medicine - Published
- 2021
77. Cardiac dysfunction in cancer patients
- Author
-
Wolfgang A. Linke, Yuri D'Alessandra, Anikó Görbe, Péter Ferdinandy, Carlo G. Tocchetti, Michele Ciccarelli, Daniela Sorriento, Serena Zacchigna, Stephane Heymans, Dana Dawson, Manuel Mayr, Nazha Hamdani, Rosalinda Madonna, Pietro Ameri, Inês Falcão-Pires, Emilio Hirsch, Rudolf A. de Boer, Alexander R. Lyon, Michele Russo, Alessandra Ghigo, Bernadett Kiss, Mauro Giacca, Jolanda van der Velden, Thomas Thum, Luc Bertrand, Tocchetti, Carlo G, Ameri, Pietro, de Boer, Rudolf A, D'Alessandra, Yuri, Russo, Michele, Sorriento, Daniela, Ciccarelli, Michele, Kiss, Bernadett, Bertrand, Luc, Dawson, Dana, Falcao-Pires, Ine, Giacca, Mauro, Hamdani, Nazha, Linke, Wolfgang A, Mayr, Manuel, van der Velden, Jolanda, Zacchigna, Serena, Ghigo, Alessandra, Hirsch, Emilio, Lyon, Alexander R, Görbe, Anikó, Ferdinandy, Péter, Madonna, Rosalinda, Heymans, Stephane, Thum, Thomas, UCL - SSS/IREC/CARD - Pôle de recherche cardiovasculaire, Cardiologie, MUMC+: MA Med Staf Spec Cardiologie (9), RS: Carim - H02 Cardiomyopathy, and Cardiovascular Centre (CVC)
- Subjects
0301 basic medicine ,RNA, Untranslated ,HISTONE DEACETYLASE ,Physiology ,heart failure ,Disease ,Cell Communication ,030204 cardiovascular system & hematology ,Gut flora ,Multicellular ,cause of death ,0302 clinical medicine ,cardiovascular disease ,Risk Factors ,inflammatory cell ,Neoplasms ,Myocytes, Cardiac ,IMMUNE CHECKPOINT INHIBITORS ,cancer fibroblasts ,antineoplastic agent ,Cause of death ,stromal cells ,toxic effect ,whole genome sequencing ,biology ,CLONAL HEMATOPOIESIS ,aging ,cardiac myocytes ,myocardium ,inflammatory cells ,cardiovascular diseases ,endothelial cells ,antineoplastic agents ,medical oncology ,heart ,cellular biology ,cardiac function ,impaired microbiome ,european society of cardiology ,EUROPEAN-SOCIETY ,3. Good health ,Cell biology ,CHAIN FATTY-ACIDS ,PRESERVED EJECTION FRACTION ,CARDIOVASCULAR-DISEASE ,endothelial cell ,Cardio-Oncology ,Inflammation Mediators ,Cardiology and Cardiovascular Medicine ,Common pathways in heart failure and cancer ,Signal Transduction ,Cardiac function curve ,Stromal cell ,Heart Diseases ,cardiac myocyte ,Antineoplastic Agents ,Risk Assessment ,03 medical and health sciences ,LUNG-CANCER ,Multicellular and multiorgan mechanisms ,Physiology (medical) ,cancer fibroblast ,medicine ,Animals ,Humans ,Lung cancer ,business.industry ,Cancer ,stromal cell ,medicine.disease ,biology.organism_classification ,Cardiotoxicity ,Gastrointestinal Microbiome ,030104 developmental biology ,Gene Expression Regulation ,Multiorgan mechanisms ,Heart failure ,INFLAMMATORY RESPONSES ,business ,DOXORUBICIN-INDUCED CARDIOTOXICITY - Abstract
In western countries, cardiovascular (CV) disease and cancer are the leading causes of death in the ageing population. Recent epidemiological data suggest that cancer is more frequent in patients with prevalent or incident CV disease, in particular, heart failure (HF). Indeed, there is a tight link in terms of shared risk factors and mechanisms between HF and cancer. HF induced by anticancer therapies has been extensively studied, primarily focusing on the toxic effects that anti-tumour treatments exert on cardiomyocytes. In this Cardio-Oncology update, members of the ESC Working Groups of Myocardial Function and Cellular Biology of the Heart discuss novel evidence interconnecting cardiac dysfunction and cancer via pathways in which cardiomyocytes may be involved but are not central. In particular, the multiple roles of cardiac stromal cells (endothelial cells and fibroblasts) and inflammatory cells are highlighted. Also, the gut microbiota is depicted as a new player at the crossroads between HF and cancer. Finally, the role of non-coding RNAs in Cardio-Oncology is also addressed. All these insights are expected to fuel additional research efforts in the field of Cardio-Oncology.
- Published
- 2020
78. Genetic lineage tracing reveals poor angiogenic potential of cardiac endothelial cells
- Author
-
Michael Rehman, Mauro Giacca, Lorena Zentilin, Nina Volf, Ralf H. Adams, Federica Benvenuti, Ambra Cappelletto, Bin Zhou, Serena Zacchigna, Elena Groppa, Giulia Maria Piperno, Tea Kocijan, Gabriele Zucca, Simone Vodret, Andrea Colliva, Matteo Leban, Kocijan, Tea, Rehman, Michael, Colliva, Andrea, Groppa, Elena, Leban, Matteo, Vodret, Simone, Volf, Nina, Zucca, Gabriele, Cappelletto, Ambra, Piperno, Giulia Maria, Zentilin, Lorena, Giacca, Mauro, Benvenuti, Federica, Bin, Zhou, Adams, Ralf, and Zacchigna, Serena
- Subjects
Vascular Endothelial Growth Factor A ,Physiology ,Angiogenesis ,Notch signaling pathway ,Neovascularization, Physiologic ,Mice, Transgenic ,Lineage tracing ,chemistry.chemical_compound ,Cell Line, Tumor ,Neoplasms ,Physiology (medical) ,Tumor Microenvironment ,medicine ,cancer ,Animals ,Cell Lineage ,Myocardial infarction ,Receptor, Notch1 ,Muscle, Skeletal ,Adaptor Proteins, Signal Transducing ,Cell Proliferation ,Mice, Inbred BALB C ,Vascular Endothelial Growth Factor Receptor-1 ,Neovascularization, Pathologic ,business.industry ,Calcium-Binding Proteins ,Editorials ,Endothelial Cells ,Skeletal muscle ,medicine.disease ,VEGF ,Coronary Vessels ,Tumor Burden ,Apelin ,Mice, Inbred C57BL ,Vascular endothelial growth factor ,Angiogenesi ,Vascular endothelial growth factor A ,Phenotype ,medicine.anatomical_structure ,Cellular Microenvironment ,chemistry ,Cancer cell ,Cancer research ,Cardiology and Cardiovascular Medicine ,business - Abstract
Aims Cardiac ischaemia does not elicit an efficient angiogenic response. Indeed, lack of surgical revascularization upon myocardial infarction results in cardiomyocyte death, scarring, and loss of contractile function. Clinical trials aimed at inducing therapeutic revascularization through the delivery of pro-angiogenic molecules after cardiac ischaemia have invariably failed, suggesting that endothelial cells in the heart cannot mount an efficient angiogenic response. To understand why the heart is a poorly angiogenic environment, here we compare the angiogenic response of the cardiac and skeletal muscle using a lineage tracing approach to genetically label sprouting endothelial cells. Methods and results We observed that overexpression of the vascular endothelial growth factor in the skeletal muscle potently stimulated angiogenesis, resulting in the formation of a massive number of new capillaries and arterioles. In contrast, response to the same dose of the same factor in the heart was blunted and consisted in a modest increase in the number of new arterioles. By using Apelin-CreER mice to genetically label sprouting endothelial cells we observed that different pro-angiogenic stimuli activated Apelin expression in both muscle types to a similar extent, however, only in the skeletal muscle, these cells were able to sprout, form elongated vascular tubes activating Notch signalling, and became incorporated into arteries. In the heart, Apelin-positive cells transiently persisted and failed to give rise to new vessels. When we implanted cancer cells in different organs, the abortive angiogenic response in the heart resulted in a reduced expansion of the tumour mass. Conclusion Our genetic lineage tracing indicates that cardiac endothelial cells activate Apelin expression in response to pro-angiogenic stimuli but, different from those of the skeletal muscle, fail to proliferate and form mature and structured vessels. The poor angiogenic potential of the heart is associated with reduced tumour angiogenesis and growth of cancer cells.
- Published
- 2020
79. Cardiac regeneration and remodelling of the cardiomyocyte cytoarchitecture
- Author
-
Hashim Ali, Mauro Giacca, Luca Braga, Ali, H., Braga, L., and Giacca, M.
- Subjects
0301 basic medicine ,Cell division ,cardiomyocyte ,Biology ,Mechanotransduction, Cellular ,Microtubules ,Biochemistry ,Sarcomere ,intercalated disc ,03 medical and health sciences ,0302 clinical medicine ,medicine ,Animals ,Humans ,Regeneration ,Myocytes, Cardiac ,Cytoskeleton ,Molecular Biology ,Transcription factor ,Actin ,Cell Proliferation ,cytoskeleton ,Cell Biology ,actin ,centrosome ,mechanosensing ,regeneration ,sarcomere ,tubulin ,YAP ,Actin cytoskeleton ,Embryonic stem cell ,Cell biology ,Actin Cytoskeleton ,030104 developmental biology ,medicine.anatomical_structure ,030220 oncology & carcinogenesis ,Intercalated disc - Abstract
Adult mammals are unable to regenerate their hearts after cardiac injury, largely due to the incapacity of cardiomyocytes (CMs) to undergo cell division. However, mammalian embryonic and fetal CMs, similar to CMs from fish and amphibians during their entire life, exhibit robust replicative activity, which stops abruptly after birth and never significantly resumes. Converging evidence indicates that formation of the highly ordered and stable cytoarchitecture of mammalian mature CMs is coupled with loss of their proliferative potential. Here, we review the available information on the role of the cardiac cytoskeleton and sarcomere in the regulation of CM proliferation. The actin cytoskeleton, the intercalated disc, the microtubular network and the dystrophin–glycoprotein complex each sense mechanical cues from the surrounding environment. Furthermore, they participate in the regulation of CM proliferation by impinging on the yes-associated protein/transcriptional co-activator with PDZ-binding motif, β-catenin and myocardin-related transcription factor transcriptional co-activators. Mastering the molecular mechanisms regulating CM proliferation would permit the development of innovative strategies to stimulate cardiac regeneration in adult individuals, a hitherto unachieved yet fundamental therapeutic goal.
- Published
- 2019
80. Functional screenings reveal different requirements for host microRNAs in Salmonella and Shigella infection
- Author
-
Malvika Sharan, Ana Eulalio, Ana Rita Cruz, Ricardo Jorge Silva, Sara Zaldívar-López, Ushasree Sunkavalli, Mauro Giacca, Claire Maudet, Ines Rodrigues Lopes, Carmen Aguilar, Clivia Lisowski, Juan J. Garrido, Miguel Mano, Aguilar, C., Cruz, A. R., Rodrigues Lopes, I., Maudet, C., Sunkavalli, U., Silva, R. J., Sharan, M., Lisowski, C., Zaldivar-Lopez, S., Garrido, J. J., Giacca, M., Mano, M., and Eulalio, A.
- Subjects
Salmonella typhimurium ,Microbiology (medical) ,Salmonella ,microRNA ,Shighella ,high throughput screening ,Swine ,Immunology ,Genomics ,Salmonella infection ,medicine.disease_cause ,Applied Microbiology and Biotechnology ,Microbiology ,Shigella flexneri ,Pathogenesis ,03 medical and health sciences ,Species Specificity ,Genetics ,medicine ,Animals ,Humans ,Shigella ,030304 developmental biology ,Regulation of gene expression ,0303 health sciences ,biology ,030306 microbiology ,Enterobacteriaceae Infections ,Cell Biology ,biology.organism_classification ,medicine.disease ,MicroRNAs ,Gene Expression Regulation ,Host-Pathogen Interactions ,HeLa Cells - Abstract
MicroRNAs (miRNAs) are increasingly recognized for their role in infection by bacterial pathogens, although the effect of each individual miRNA remains largely unknown. Here, we used a comparative genome-wide microscopy-based functional screening approach to identify miRNAs controlling infection by two bacterial pathogens-Salmonella enterica serovar Typhimurium and Shigella flexneri. Despite the similarities between these pathogens, we found infections to be controlled by largely non-overlapping subsets of miRNAs, seemingly reflecting different requirements prompted by their distinct intracellular lifestyles. By characterizing a small subset of miRNAs chosen among the strongest inhibitors of Shigella infection, we discovered that miR-3668, miR-4732-5p and miR-6073 exert a selective effect on Shigella infection by impairing bacterial actin-based motility by downregulating N-WASP. Additionally, by identifying let-7i-3p miRNA as a strong inhibitor of Salmonella replication and performing in-depth analysis of its mechanisms of action, we showed that this miRNA specifically inhibits Salmonella infection via modulation of endolysosomal trafficking and the vacuolar environment by targeting the host RGS2 protein. These findings illustrate two paradigms underlying miRNA-mediated regulation of bacterial infection, acting as part of the host response to infection, or as part of bacterial strategies to modulate the host environment and favour pathogenesis.
- Published
- 2019
81. SARS-CoV-2 Spike protein activates TMEM16F-mediated platelet pro-coagulant activity
- Author
-
Ambra Cappelletto, Harriet E. Allan, Marilena Crescente, Edoardo Schneider, Rossana Bussani, Hashim Ali, Ilaria Secco, Simone Vodret, Roberto Simeone, Luca Mascaretti, Serena Zacchigna, Timothy D. Warner, and Mauro Giacca
- Subjects
Platelets ,Coagulation ,SARS-CoV-2 ,TMEM16F ,Niclosamide ,Clofazimine - Abstract
BackgroundThrombosis of the lung micro-vasculature is a characteristic of COVID-19 disease, which is observed in large excess compared to other forms of acute respiratory distress syndrome and thus suggests a trigger for thrombosis endogenous to the lung. Our recent work has shown that the SARS-CoV-2 Spike protein activates the cellular TMEM16F chloride channel and scramblase. Through a screening on >3,000 FDA/EMA approved drugs, we identified Niclosamide and Clofazimine as the most effective molecules at inhibiting this activity. As TMEM16F plays an important role in the stimulation of the pro-coagulant activity of platelets, and considering that platelet abnormalities are common in COVID-19 patients, we investigated whether Spike directly affects platelet activation and pro-thrombotic function and tested the effect of Niclosamide and Clofazimine on these processes.MethodsWe produced SARS-CoV-2 Spike or VSV-G protein-pseudotyped virions, or generated cells expressing Spike on their plasma membrane, and tested their effects on platelet adhesion (fluorescence), aggregation (absorbance), exposure of phosphatidylserine (flow cytometry for annexin V binding), calcium flux (flow cytometry for fluo-4 AM), and clot formation and retraction. These experiments were also conducted in the presence of the TMEM16F activity inhibitors Niclosamide and Clofazimine.ResultsHere we show that exposure to SARS-CoV-2 Spike promotes platelet activation, adhesion and spreading, both when present on the envelope of virions or upon expression on the plasma membrane of cells. Spike was effective both as a sole agonist or by enhancing the effect of known platelet activators, such as collagen and collagen-related peptide. In particular, Spike exerted a noticeable effect on the procoagulant phenotype of platelets, by enhancing calcium flux, phosphatidylserine externalisation, and thrombin generation. Eventually, this resulted in a striking increase in thrombin-induced clot formation and retraction. Both Niclosamide and Clofazimine almost abolished this Spike-induced pro-coagulant response.ConclusionsTogether, these findings provide a pathogenic mechanism to explain thrombosis associated to COVID-19 lung disease, by which Spike present in SARS-CoV-2 virions or exposed on the surface of infected cells, leads to local platelet stimulation and subsequent activation of the coagulation cascade. As platelet TMEM16F is central in this process, these findings reinforce the rationale of repurposing drugs targeting this protein, such as Niclosamide, for COVID-19 therapy.
- Published
- 2021
82. Cardiomyocytes stimulate angiogenesis after ischemic injury in a ZEB2-dependent manner
- Author
-
Mauro Giacca, Eva van Rooij, Danny Huylebroeck, Manon M. H. Huibers, Jody J. Haigh, D Versteeg, Reinier A. Boon, Bas Molenaar, Monika M Gladka, Veerle Kremer, Harmjan R. Vos, Arwa Kohela, Lieneke Kooijman, Jantine Monshouwer-Kloots, AGEM - Amsterdam Gastroenterology Endocrinology Metabolism, ACS - Atherosclerosis & ischemic syndromes, ACS - Diabetes & metabolism, Graduate School, ACS - Heart failure & arrhythmias, Cell biology, Hubrecht Institute for Developmental Biology and Stem Cell Research, Physiology, and ACS - Microcirculation
- Subjects
0301 basic medicine ,RNA, Messenger/genetics ,Myocardial Infarction/genetics ,Angiogenesis ,Physiologic/genetics ,Dependovirus/metabolism ,Myocardial Infarction ,General Physics and Astronomy ,Infarction ,Neovascularization ,Mice ,0302 clinical medicine ,Models ,Cell Movement ,Ischemia ,Myocyte ,Myocytes, Cardiac ,Thymosin/analogs & derivatives ,Mice, Knockout ,Multidisciplinary ,Myocytes, Cardiac/metabolism ,Dependovirus ,Cell biology ,Endothelial stem cell ,030220 oncology & carcinogenesis ,Cardiac/metabolism ,medicine.symptom ,Cardiac function curve ,Science ,Knockout ,Neovascularization, Physiologic ,Models, Biological ,General Biochemistry, Genetics and Molecular Biology ,Article ,03 medical and health sciences ,Paracrine signalling ,Cell Proliferation/genetics ,medicine ,Cell Movement/genetics ,Animals ,Humans ,RNA, Messenger ,Protein Precursors ,Cell Proliferation ,Zinc Finger E-box Binding Homeobox 2 ,Myocytes ,business.industry ,Protein Precursors/genetics ,General Chemistry ,medicine.disease ,Biological ,Zinc Finger E-box Binding Homeobox 2/genetics ,Thymosin ,030104 developmental biology ,Gene Expression Regulation ,RNA ,Messenger/genetics ,Ischemia/genetics ,business ,Neovascularization, Physiologic/genetics - Abstract
The disruption in blood supply due to myocardial infarction is a critical determinant for infarct size and subsequent deterioration in function. The identification of factors that enhance cardiac repair by the restoration of the vascular network is, therefore, of great significance. Here, we show that the transcription factor Zinc finger E-box-binding homeobox 2 (ZEB2) is increased in stressed cardiomyocytes and induces a cardioprotective cross-talk between cardiomyocytes and endothelial cells to enhance angiogenesis after ischemia. Single-cell sequencing indicates ZEB2 to be enriched in injured cardiomyocytes. Cardiomyocyte-specific deletion of ZEB2 results in impaired cardiac contractility and infarct healing post-myocardial infarction (post-MI), while cardiomyocyte-specific ZEB2 overexpression improves cardiomyocyte survival and cardiac function. We identified Thymosin β4 (TMSB4) and Prothymosin α (PTMA) as main paracrine factors released from cardiomyocytes to stimulate angiogenesis by enhancing endothelial cell migration, and whose regulation is validated in our in vivo models. Therapeutic delivery of ZEB2 to cardiomyocytes in the infarcted heart induces the expression of TMSB4 and PTMA, which enhances angiogenesis and prevents cardiac dysfunction. These findings reveal ZEB2 as a beneficial factor during ischemic injury, which may hold promise for the identification of new therapies., ZEB2 transcription factor is increased in a subset of cardiomyocytes during stress to induce cardioprotective effects after injury. Here the authors show that therapeutic delivery of ZEB2 prevents cardiac dysfunction after ischemic damage and promotes the activation of pro-angiogenic signals.
- Published
- 2021
83. CycleTrack, a genetic method to visualize cardiomyocyte renewal in vivo
- Author
-
Ilaria Secco, Ana Backovic, Simone Vodret, Matteo Dal Ferro, Mateusz Tomczyk, Khatia Gabisonia, Lucia Carlucci, Lorena Zentilin, Serena Zacchigna, Fabio A. Recchia, and Mauro Giacca
- Subjects
Cardiology and Cardiovascular Medicine ,Molecular Biology - Published
- 2022
84. Lipid nanoparticles for cardiac RNA delivery
- Author
-
Josef Huntington, Hend Mohamed Abdel-Bar, Manendra Pachauri, Mateusz Tomczyk, Yue Qin, Luca Venditti, Khuloud Al-Jamal, and Mauro Giacca
- Subjects
Cardiology and Cardiovascular Medicine ,Molecular Biology - Published
- 2022
85. Dyrk1a kinase controls cardiomyocyte proliferation
- Author
-
Giorgia Rizzari, Ellen Dirkx, Consuelo Torrini, Mateusz Tomczyk, Miguel Mano, and Mauro Giacca
- Subjects
Cardiology and Cardiovascular Medicine ,Molecular Biology - Published
- 2022
86. I06 SREBP2 delivery to striatal astrocytes normalizes transcription of cholesterol biosynthesis genes and ameliorates pathological features in huntington’s disease
- Author
-
Gianluca Verlengia, Francesca Talpo, Chiara Cordiglieri, Gerardo Biella, Giulia Birolini, Michele Simonato, Mauro Giacca, Paola Conforti, Franco Taroni, Claudia Maniezzi, Marta Valenza, Valerio Leoni, Elena Cattaneo, Lorena Zentilin, and Claudio Caccia
- Subjects
Huntingtin ,Cholesterol ,Endogeny ,Biology ,medicine.disease ,Cell biology ,Pathogenesis ,chemistry.chemical_compound ,Huntington's disease ,chemistry ,Transcription (biology) ,medicine ,Huntingtin Protein ,Transcription factor - Abstract
Background Cholesterol is a multifaceted molecule essential for brain function (Martin 2014). In the adult brain, cholesterol is produced locally by astrocytes and transferred to neurons through apoE-containing lipoproteins (Jurevics & Morell 1995; Mauch 2001). Disruption of brain cholesterol pathways has been linked to several neurological disorders, including Huntington’s disease (HD), a genetic, neurodegenerative disorder caused by a CAG expansion in the gene encoding the Huntingtin protein (Valenza & Cattaneo 2011). Brain cholesterol biosynthesis and content are reduced in several HD models (Valenza 2005; 2007; 2010; Shankaran 2017). The underlying molecular mechanism relies on reduced nuclear translocation of SREBP2, the transcription factor that controls the transcription of several genes involved in cholesterol biosynthesis (Valenza 2015; Di Pardo 2020). We have recently shown that cholesterol supplementation to the brain, with different delivery systems, ameliorates synaptic and behavioral defects in the R6/2 mouse model (Valenza 2015; Birolini 2020; Birolini 2021). Aims and Methods Here, we used recombinant adeno-associated virus 2/5 to deliver exogenous SREBP2 specifically in astrocytes in order to enhance the endogenous cholesterol biosynthesis in the striatum of R6/2 mice. Results We found that exogenous SREBP2 stimulates the transcription of key cholesterol biosynthesis genes resulting in fully restoration of synaptic transmission, reversal of Drd2 transcript levels, clearance of mutant Huntingtin (muHTT) aggregates and rescue of behavioral deficits. Conclusions These results demonstrate that stimulating cholesterol biosynthesis in striatal astrocytes has a positive effect on behavioral decline and disease-related phenotypes in HD mice. Furthermore, we have demonstrated that glial SREBP2 participates in HD pathogenesis in vivo, highlighting the translational potential of cholesterol-based strategies for this disease.
- Published
- 2021
87. A microRNA program regulates the balance between cardiomyocyte hyperplasia and hypertrophy and stimulates cardiac regeneration
- Author
-
Vasco Sampaio-Pinto, Lara Ottaviani, Luca Braga, Roel A. de Weger, Leon J. De Windt, Lorena Zentillin, Daniel W. Sorensen, Jop H. van Berlo, Andrea Raso, Serena Zacchigna, Silvia Moimas, Paula A. da Costa Martins, Diana S. Nascimento, Manon M. H. Huibers, Hamid el Azzouzi, Ellen Dirkx, Sailay Siddiqi, Ryan John Cubero, Servé Olieslagers, Consuelo Torrini, Mauro Giacca, Raso, A., Dirkx, E., Sampaio-Pinto, V., el Azzouzi, H., Cubero, R. J., Sorensen, D. W., Ottaviani, L., Olieslagers, S., Huibers, M. M., de Weger, R., Siddiqi, S., Moimas, S., Torrini, C., Zentillin, L., Braga, L., Nascimento, D. S., da Costa Martins, P. A., van Berlo, J. H., Zacchigna, S., Giacca, M., De Windt, L. J., RS: Carim - Heart, Cardiologie, RS: FSE DMG, RS: Carim - H05 Gene regulation, and Molecular Genetics
- Subjects
Myocardial Infarction ,General Physics and Astronomy ,Muscle hypertrophy ,ACTIVATION ,PATHWAY ,Mice ,Myocyte ,Myocytes, Cardiac ,Cells, Cultured ,IN-VIVO ,Regulation of gene expression ,Multidisciplinary ,Cultured ,biology ,Reverse Transcriptase Polymerase Chain Reaction ,PROLIFERATION ,food and beverages ,MicroRNA ,Hyperplasia ,Cell cycle ,Cell biology ,Echocardiography ,miRNAs ,Cardiac regeneration ,HAND2 ,Cardiac ,Human ,HEART REGENERATION ,EXPRESSION ,Science ,Cells ,Cardiomegaly ,Gene delivery ,MYOCYTES ,Article ,General Biochemistry, Genetics and Molecular Biology ,ADULT ,REGRESSION ,medicine ,Animals ,Humans ,Regeneration ,Animals, Newborn ,Gene Expression Regulation ,MicroRNAs ,Rats ,Animal ,Regeneration (biology) ,General Chemistry ,medicine.disease ,Newborn ,ATRIAL-FIBRILLATION ,biology.protein ,Rat - Abstract
Myocardial regeneration is restricted to early postnatal life, when mammalian cardiomyocytes still retain the ability to proliferate. The molecular cues that induce cell cycle arrest of neonatal cardiomyocytes towards terminally differentiated adult heart muscle cells remain obscure. Here we report that the miR-106b~25 cluster is higher expressed in the early postnatal myocardium and decreases in expression towards adulthood, especially under conditions of overload, and orchestrates the transition of cardiomyocyte hyperplasia towards cell cycle arrest and hypertrophy by virtue of its targetome. In line, gene delivery of miR-106b~25 to the mouse heart provokes cardiomyocyte proliferation by targeting a network of negative cell cycle regulators including E2f5, Cdkn1c, Ccne1 and Wee1. Conversely, gene-targeted miR-106b~25 null mice display spontaneous hypertrophic remodeling and exaggerated remodeling to overload by derepression of the prohypertrophic transcription factors Hand2 and Mef2d. Taking advantage of the regulatory function of miR-106b~25 on cardiomyocyte hyperplasia and hypertrophy, viral gene delivery of miR-106b~25 provokes nearly complete regeneration of the adult myocardium after ischemic injury. Our data demonstrate that exploitation of conserved molecular programs can enhance the regenerative capacity of the injured heart., Myocardial regeneration and proliferation of heart muscle cells is limited to a short period after birth early postnatal life, after which heart muscle cells can only grow in size and not in number. Here, the authors identified that the expression level of an endogenous microRNA cluster in heart muscle promotes the passage of the proliferative state to adult heart growth, and modulating the expression of this cluster can stimulate heart regeneration after myocardial infarction.
- Published
- 2021
88. Impact of the COVID-19 pandemic on in-hospital mortality in cardiovascular disease: a meta-analysis
- Author
-
Daniel I. Bromage, Allen Daniel, Samuel A. Watson, Mauro Giacca, Theresa McDonagh, Antonio Cannatà, Paul A. Scott, and Ajay M. Shah
- Subjects
medicine.medical_specialty ,Coronavirus disease 2019 (COVID-19) ,Epidemiology ,Disease ,030204 cardiovascular system & hematology ,Full Research Paper ,03 medical and health sciences ,0302 clinical medicine ,Internal medicine ,Pandemic ,Medicine ,Humans ,AcademicSubjects/MED00200 ,030212 general & internal medicine ,Hospital Mortality ,Pandemics ,business.industry ,SARS-CoV-2 ,Mortality rate ,COVID-19 ,Cardiovascular disease ,Confidence interval ,In-hospital mortality ,Meta-analysis ,Cardiovascular Diseases ,Relative risk ,Cohort ,Cardiology and Cardiovascular Medicine ,business - Abstract
Aims The COVID-19 pandemic has resulted in excess mortality due to both COVID-19 directly and other conditions, including cardiovascular (CV) disease. We aimed to explore the excess in-hospital mortality, unrelated to COVID-19 infection, across a range of CV diseases. Methods and results A systematic search was performed for studies investigating in-hospital mortality among patients admitted with CV disease without SARS-CoV-2 infection compared with a period outside the COVID-19 pandemic. Fifteen studies on 27 421 patients with CV disease were included in the analysis. The average in-hospital mortality rate was 10.4% (n = 974) in the COVID-19 group and 5.7% (n = 1026) in the comparator group. Compared with periods outside the COVID-19 pandemic, the pooled risk ratio (RR) demonstrated increased in-hospital mortality by 62% during COVID-19 [95% confidence interval (CI) 1.20–2.20, P = 0.002]. Studies with a decline in admission rate >50% during the COVID-19 pandemic observed the greatest increase in mortality compared with those with Conclusions In-hospital mortality among patients admitted with CV diseases was increased relative to periods outside the pandemic, independent of co-infection with COVID-19. This effect was larger in studies with the biggest decline in admission rates, suggesting a sicker cohort of patients in this period. However, studies were generally poorly conducted, and there is a need for further well-designed studies to establish the full extent of mortality not directly related to COVID-19 infection.
- Published
- 2021
89. High Throughput miRNA Screening Identifies miR-574-3p Hyperproductive Effect in CHO Cells
- Author
-
Natasa Skoko, Corrado Guarnaccia, Marco Baralle, Jeremias S. Herzog, Živa Švab, Luca Braga, Ivan Labik, and Mauro Giacca
- Subjects
0106 biological sciences ,p53 ,miR-574-3p ,CHO ,Cell ,Endogeny ,p300 ,Computational biology ,CHO Cells ,Microbiology ,01 natural sciences ,Biochemistry ,Chinese hamster ,Article ,Etanercept ,03 medical and health sciences ,Cricetulus ,010608 biotechnology ,microRNA ,medicine ,hyperproductivity ,Animals ,Humans ,Transgenes ,Molecular Biology ,Erythropoietin ,030304 developmental biology ,0303 health sciences ,Gene knockdown ,biology ,Chinese hamster ovary cell ,biology.organism_classification ,QR1-502 ,Recombinant Proteins ,High-Throughput Screening Assays ,MicroRNAs ,medicine.anatomical_structure ,Gene Expression Regulation ,Cell culture ,miRNA screening ,Nucleic Acid Conformation ,Tumor Suppressor Protein p53 ,Genetic Engineering ,E1A-Associated p300 Protein ,pri-miRNA processing ,medicine.drug - Abstract
CHO is the cell line of choice for the manufacturing of many complex biotherapeutics. The constant upgrading of cell productivity is needed to meet the growing demand for these life-saving drugs. Manipulation of small non-coding RNAs—miRNAs—is a good alternative to a single gene knockdown approach due to their post-transcriptional regulation of entire cellular pathways without posing translational burden to the production cell. In this study, we performed a high-throughput screening of 2042-human miRNAs and identified several candidates able to increase cell-specific and overall production of Erythropoietin and Etanercept in CHO cells. Some of these human miRNAs have not been found in Chinese hamster cells and yet were still effective in them. We identified miR-574-3p as being able, when overexpressed in CHO cells, to improve overall productivity of Erythropoietin and Etanercept titers from 1.3 to up to 2-fold. In addition, we validated several targets of miR-574-3p and identified p300 as a main target of miR-574-3p in CHO cells. Furthermore, we demonstrated that stable CHO cell overexpressing miRNAs from endogenous CHO pri-miRNA sequences outperform the cells with human pri-miRNA sequences. Our findings highlight the importance of flanking genomic sequences, and their secondary structure features, on pri-miRNA processing offering a novel, cost-effective and fast strategy as a valuable tool for efficient miRNAs engineering in CHO cells.
- Published
- 2021
90. SARS-CoV-2 RNAemia and proteomic trajectories inform prognostication in COVID-19 patients admitted to intensive care
- Author
-
Ella Reed, Clemens Gutmann, Bhawana Singh, Torsten Tonn, Sean A. Burnap, Salvatore Napoli, Ajay M. Shah, Jonathan D. Edgeworth, Manu Shankar-Hari, Konstantinos Theofilatos, Kaloyan Takov, Lukas E Schmidt, Kevin O'Gallagher, Katie J. Doores, Mauro Giacca, Rafael Fernandez-Leiro, Konstantina Dimitrakopoulou, Barnaby Sanderson, Mansoor Saqi, Mark J. W. McPhail, Adrian Hayday, Hashim Ali, Salma F Mujib, Georg Auzinger, Matthew Fish, Romy Kronstein-Wiedemann, Manuel Mayr, Francesca Trovato, Marieke Rienks, Stefan R. Bornstein, Christian Cassel, Umar Niazi, Adam Nabeebaccus, Silke Braun, Blair Merrick, Maria Hasman, Xiaoke Yin, Gutmann, Clemens [0000-0003-0675-8632], Takov, Kaloyan [0000-0002-8642-6306], Burnap, Sean A [0000-0002-3408-8608], Singh, Bhawana [0000-0002-5834-8320], Theofilatos, Konstantinos [0000-0001-6799-0553], Fish, Matthew [0000-0001-6462-3889], Schmidt, Lukas E [0000-0001-7565-1455], Rienks, Marieke [0000-0002-0590-9518], Yin, Xiaoke [0000-0002-5172-0935], Sanderson, Barnaby [0000-0002-9621-143X], Merrick, Blair [0000-0002-6061-6064], Niazi, Umar [0000-0001-7176-8883], Fernández-Leiro, Rafael [0000-0002-7941-0357], Braun, Silke [0000-0002-2558-1521], Doores, Katie J [0000-0002-5507-1725], Bornstein, Stefan R [0000-0002-5211-2536], Tonn, Torsten [0000-0001-9580-2193], Hayday, Adrian C [0000-0002-9495-5793], Giacca, Mauro [0000-0003-2927-7225], Shankar-Hari, Manu [0000-0002-5338-2538], Mayr, Manuel [0000-0002-0597-829X], and Apollo - University of Cambridge Repository
- Subjects
0301 basic medicine ,Proteomics ,Male ,General Physics and Astronomy ,Kaplan-Meier Estimate ,law.invention ,Prognostic markers ,0302 clinical medicine ,law ,Medicine ,Mannan-binding lectin ,Serum Amyloid P-Component/metabolism ,Multidisciplinary ,biology ,COVID-19/metabolism ,Antigens, Neoplasm/metabolism ,Hazard ratio ,PTX3 ,Viral Load ,Middle Aged ,Intensive care unit ,Antibodies, Neutralizing/immunology ,Serum Amyloid P-Component ,C-Reactive Protein ,030220 oncology & carcinogenesis ,Spike Glycoprotein, Coronavirus ,RNA, Viral ,Female ,Antibody ,Proteomics/methods ,Adult ,medicine.medical_specialty ,Critical Care ,Science ,General Biochemistry, Genetics and Molecular Biology ,Article ,Sepsis ,03 medical and health sciences ,Antigens, Neoplasm ,SARS-CoV-2/genetics ,Internal medicine ,Intensive care ,Biomarkers, Tumor ,Animals ,Humans ,RNA, Viral/blood ,Mass spectrometry ,business.industry ,SARS-CoV-2 ,C-Reactive Protein/metabolism ,Viral Load/immunology ,Critical Care/statistics & numerical data ,COVID-19 ,General Chemistry ,medicine.disease ,Antibodies, Neutralizing ,Complement system ,030104 developmental biology ,HEK293 Cells ,Biomarkers, Tumor/metabolism ,biology.protein ,Spike Glycoprotein, Coronavirus/immunology ,business - Abstract
Prognostic characteristics inform risk stratification in intensive care unit (ICU) patients with coronavirus disease 2019 (COVID-19). We obtained blood samples (n = 474) from hospitalized COVID-19 patients (n = 123), non-COVID-19 ICU sepsis patients (n = 25) and healthy controls (n = 30). Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RNA was detected in plasma or serum (RNAemia) of COVID-19 ICU patients when neutralizing antibody response was low. RNAemia is associated with higher 28-day ICU mortality (hazard ratio [HR], 1.84 [95% CI, 1.22–2.77] adjusted for age and sex). RNAemia is comparable in performance to the best protein predictors. Mannose binding lectin 2 and pentraxin-3 (PTX3), two activators of the complement pathway of the innate immune system, are positively associated with mortality. Machine learning identified ‘Age, RNAemia’ and ‘Age, PTX3’ as the best binary signatures associated with 28-day ICU mortality. In longitudinal comparisons, COVID-19 ICU patients have a distinct proteomic trajectory associated with mortality, with recovery of many liver-derived proteins indicating survival. Finally, proteins of the complement system and galectin-3-binding protein (LGALS3BP) are identified as interaction partners of SARS-CoV-2 spike glycoprotein. LGALS3BP overexpression inhibits spike-pseudoparticle uptake and spike-induced cell-cell fusion in vitro., Here the authors use RT-qPCR and mass spectrometry to analyze longitudinal blood samples from intensive care unit (ICU) COVID-19 patients and controls. They find that viral RNA and pentraxin-3 predict 28-day ICU mortality and that galectin-3-binding protein is an interaction partner of SARS-CoV-2 spike glycoprotein with antiviral properties.
- Published
- 2021
91. BS25 TAM receptor AXL loss regulates smooth muscle cell differentiation and accelerates atherosclerosis in mice
- Author
-
Thomas D. Otto, Lucy McShane, Danila Gurgone, Gabrielle Paulsson-Berne, Gianluca Grassia, Mariola Kurowska-Stolarska, Neil MacRitchie, Abdullatif Mohammed Bin-Khunayn, Glykeria Karadimou, Pasquale Maffia, Eva Crespo, Tomasz J. Guzik, Peder S. Olofsson, Lorena Zentilin, and Mauro Giacca
- Subjects
biology ,business.industry ,Smooth muscle cell differentiation ,Inflammation ,MERTK ,Receptor tyrosine kinase ,Vascular remodelling in the embryo ,Cancer research ,biology.protein ,Medicine ,medicine.symptom ,business ,Efferocytosis ,Receptor ,TYRO3 - Abstract
Introduction The TAM receptors (Tyro3, Axl, and MerTK) are a distinct family of three receptor tyrosine kinases, namely Tyro3, Axl and MerTK, which play critical roles in cancer, inflammatory disorders and cardiovascular diseases. Axl, in particular, has been shown to influence multiple aspects of cardiovascular pathology via diverse effects on cells of both the vasculature and immune system through regulation of vascular remodelling, efferocytosis and inflammation. Clinical studies have shown that Axl is detectable in atherosclerotic plaques; however, the causal relationship between Axl and atherosclerosis is still uncertain, and results from mouse models fell short of defining the specific role(s) of Axl in the disease process. Methods In order to quantify Axl expression in carotid endarterectomy atherosclerotic plaques we examined data from the Biobank of Karolinska Endarterectomy (BiKE). Using single-cell RNA sequencing (scRNA-seq) data from published atherosclerosis datasets we determined which cell types express Axl during pathology. Finally, we utilised an inducible atherosclerosis model in order to assess atherosclerosis formation in global Axl-deficient mice (Axl-/-). Female C57BL/6NQ (WT; n=15) and Axl-/- (n=21) mice were injected with 3×1011 vector genomes of AAV8-proprotein convertase subtilisin/kexin type 9 (PCSK9) and placed on a Western Diet (WD) for 12 weeks. Plaque size and percentage of necrotic core were determined in the aortic sinus using Oil Red O (ORO). Collagen content was determined using picrosirius red and polarised light microscopy. ScRNA-seq was performed to explore differences in Axl-/- vs. WT aortas at the cellular and molecular level. Results We found expression of Axl in human carotid plaque to be significantly reduced in comparison to healthy control tissue (P=1.96e-06) in the BiKE cohort. Similarly, we detected less Axl RNA expression in the aortas of WD-fed apolipoprotein-E-/- mice compared to WT (P Conclusions In conclusion, our results indicate a protective role for Axl in atherosclerosis. The TAM receptor is reduced in diseased vessel compared to healthy controls in both human and mouse. Furthermore, global knock-out resulted in significantly increased plaque burden in mice. The necrotic core was not found to be influenced by Axl, suggesting that TAM receptor-mediated efferocytosis is not a key contributor to the role of Axl in atherosclerosis. Axl was found to be predominantly expressed in the VSMC compartment in the aortas of both healthy and diseased mice. Furthermore, Axl deficiency promoted VSMC phenotypic switching. These data support the hypothesis of a beneficial role of Axl in atherosclerosis via modulation of smooth muscle cell phenotype. Conflict of Interest none
- Published
- 2021
92. The furin cleavage site in the SARS-CoV-2 spike protein is required for transmission in ferrets
- Author
-
Laury Baillon, Rebecca Frise, Jack A. Hassard, Thomas P. Peacock, Mauro Giacca, Brian Hanley, Wendy S. Barclay, Jonathan Brown, Olivia C. Swann, Raul Y. Sanchez-David, David A. Matthews, Rebecca Penn, Luca Braga, Michael Osborn, Ruthiran Kugathasan, Ecco Staller, Andrew D. Davidson, Maia Kavanagh Williamson, Jie Zhou, and Daniel H. Goldhill
- Subjects
INFLUENZA-VIRUS ,viruses ,Respiratory System ,HOST-CELL ENTRY ,Virus Replication ,Applied Microbiology and Biotechnology ,ACTIVATION ,Viral genome packaging ,RESPIRATORY SYNDROME CORONAVIRUS ,1108 Medical Microbiology ,INFECTION ,Chlorocebus aethiops ,Furin ,chemistry.chemical_classification ,0303 health sciences ,biology ,viral transmission ,Serine Endopeptidases ,RNA-Binding Proteins ,Covid19 ,respiratory system ,Cell biology ,Virus Shedding ,Spike Glycoprotein, Coronavirus ,Life Sciences & Biomedicine ,0605 Microbiology ,Microbiology (medical) ,EXPRESSION ,Immunology ,Endosomes ,Cleavage (embryo) ,Microbiology ,Virus ,03 medical and health sciences ,Viral entry ,Genetics ,Animals ,Humans ,PROTEOLYTIC CLEAVAGE ,Vero Cells ,TMPRSS2 ,030304 developmental biology ,Immune Evasion ,Science & Technology ,virus-host interaction ,030306 microbiology ,SARS-CoV-2 ,fungi ,Ferrets ,COVID-19 ,Membrane Proteins ,Epithelial Cells ,Viral Genome Packaging ,Cell Biology ,Virus Internalization ,Cathepsins ,respiratory tract diseases ,Viral replication ,chemistry ,Vero cell ,biology.protein ,Glycoprotein - Abstract
SARS-CoV-2 entry requires sequential cleavage of the spike glycoprotein at the S1/S2 and the S2' cleavage sites to mediate membrane fusion. SARS-CoV-2 has a polybasic insertion (PRRAR) at the S1/S2 cleavage site that can be cleaved by furin. Using lentiviral pseudotypes and a cell-culture-adapted SARS-CoV-2 virus with an S1/S2 deletion, we show that the polybasic insertion endows SARS-CoV-2 with a selective advantage in lung cells and primary human airway epithelial cells, but impairs replication in Vero E6, a cell line used for passaging SARS-CoV-2. Using engineered spike variants and live virus competition assays and by measuring growth kinetics, we find that the selective advantage in lung and primary human airway epithelial cells depends on the expression of the cell surface protease TMPRSS2, which enables endosome-independent virus entry by a route that avoids antiviral IFITM proteins. SARS-CoV-2 virus lacking the S1/S2 furin cleavage site was shed to lower titres from infected ferrets and was not transmitted to cohoused sentinel animals, unlike wild-type virus. Analysis of 100,000 SARS-CoV-2 sequences derived from patients and 24 human postmortem tissues showed low frequencies of naturally occurring mutants that harbour deletions at the polybasic site. Taken together, our findings reveal that the furin cleavage site is an important determinant of SARS-CoV-2 transmission.
- Published
- 2021
93. VEGF-B Gene Therapy for the Heart: Proceed with Caution
- Author
-
Fabio A. Recchia and Mauro Giacca
- Subjects
Pharmacology ,0303 health sciences ,biology ,Extramural ,business.industry ,Genetic enhancement ,VEGF receptors ,MEDLINE ,030204 cardiovascular system & hematology ,Bioinformatics ,Genetic therapy ,03 medical and health sciences ,0302 clinical medicine ,Text mining ,Drug Discovery ,Genetics ,biology.protein ,Molecular Medicine ,Medicine ,business ,Molecular Biology ,030304 developmental biology - Published
- 2020
94. MicroRNA hsa-miR-29a-3p is a plasma biomarker for the differential diagnosis and monitoring of tuberculosis
- Author
-
Elvis Ndukong Ndzi, Céline Nguefeu Nkenfou, Ousman Tamgue, Jules-Roger Kuiate, Mauro Giacca, Eric Walter Yone Pefura, Linda Chapdeleine Mouafo Mekue, Alexis Ndjolo, Lorena Zentilin, Ndzi, E. N., Nkenfou, C. N., Mekue, L. M., Zentilin, L., Tamgue, O., Pefura, E. W. Y., Kuiate, J. -R., Giacca, M., and Ndjolo, A.
- Subjects
Male ,0301 basic medicine ,Oncology ,Biomarker ,Diagnosis ,microRNA ,Pediatric/extra-pulmonary tuberculosis ,Performance ,Adolescent ,Adult ,Aged ,Antitubercular Agents ,Biomarkers ,CD4 Lymphocyte Count ,Case-Control Studies ,Coinfection ,Diagnosis, Differential ,Drug Administration Schedule ,Female ,Gene Expression Profiling ,HIV Infections ,Humans ,Latent Tuberculosis ,MicroRNAs ,Middle Aged ,Tuberculosis ,Up-Regulation ,Young Adult ,Antitubercular Agent ,HIV Infection ,Latent Tuberculosi ,Latent tuberculosis ,Infectious Diseases ,Biomarker (medicine) ,medicine.symptom ,Case-Control Studie ,Diagnosi ,Human ,Microbiology (medical) ,medicine.medical_specialty ,Tuberculosi ,030106 microbiology ,Immunology ,Microbiology ,03 medical and health sciences ,Internal medicine ,medicine ,Pediatric/extra-pulmonary tuberculosi ,Receiver operating characteristic ,business.industry ,Case-control study ,medicine.disease ,030104 developmental biology ,Differential ,Sputum ,Differential diagnosis ,business - Abstract
The diagnosis of tuberculosis (TB) continues to pose substantial public health problems. The quest for diagnostic biomarkers for TB is therefore primordial. This study aimed to evaluate the diagnostic and anti-TB treatment monitoring potentials of some selected miRNAs. Quantitative real time polymerase chain reaction and Receiver operating characteristics were used to estimate the ability of miRNAs to discriminate between healthy controls (HEC), latent (LTB) and active TB (ATB). The study showed that: hsa-miR-29a-3p, hsa-miR-155-5p and hsa-miR-361-5p were significantly upregulated in ATB compared to HEC while hsa-miR-29a-3p, and hsa-miR-361-5p were also significantly up-regulated in ATB compared to LTB (all P ≤ 0.05). MiR-29a-3p showed a good (81.37%) distinguishing performance in discriminating ATB from HEC and a good (84.35%) diagnostic performance in discriminating ATB from LTB. The performance of miR-29a-3p present in the blood in discriminating active TB from latent TB and healthy controls indicates it may be a useful biomarker for diagnosis of TB. Because this miRNA is found in blood (plasma) which is easy to collect compared to sputum it could be used in pediatric and extra-pulmonary TB cases.
- Published
- 2019
95. 3D Carbon-Nanotube-Based Composites for Cardiac Tissue Engineering
- Author
-
Gabriele Baj, Valentina Martinelli, Orfeo Sbaizero, Carlin S. Long, Maurizio Prato, Luisa Mestroni, Mauro Giacca, Brisa Peña, Susanna Bosi, Martinelli, Valentina, Bosi, Susanna, Peña, Brisa, Baj, Gabriele, Long, Carlin S., Sbaizero, Orfeo, Giacca, Mauro, Prato, Maurizio, and Mestroni|, Luisa
- Subjects
0301 basic medicine ,Scaffold ,Cellular differentiation ,nanoscaffolds ,Carbon nanotubes ,Biomedical Engineering ,cardiomyocytes ,cardiomyocyte ,02 engineering and technology ,Carbon nanotube ,calcium signaling ,law.invention ,Biomaterials ,03 medical and health sciences ,chemistry.chemical_compound ,Calcium imaging ,Tissue engineering ,PDMS ,law ,medicine ,Myocyte ,Composite material ,tissue engineering ,cardiac repair ,Polydimethylsiloxane ,Biochemistry (medical) ,General Chemistry ,021001 nanoscience & nanotechnology ,medicine.disease ,030104 developmental biology ,chemistry ,Heart failure ,0210 nano-technology ,nanoscaffold - Abstract
Heart failure is a disease of epidemic proportion, and is a leading cause of mortality in the world. Since cardiac myocytes are terminally differentiated cells with minimal intrinsic ability to self-regenerate, cardiac tissue engineering has emerged as one of the most realistic therapeutic strategies for cardiac repair. We have previously proven the ability of carbon nanotube scaffolds to promote cardiomyocytes proliferation, maturation and long-term survival. Here, we tested if 3-dimensional scaffolds of carbon nanotube-based composites can also promote cardiomyocyte growth, electrophysiological maturation, and formation of functional syncytia. To this purpose, we developed an elastomeric scaffold which consists of a microporous and self–standing material made of polydimethylsiloxane (PDMS) containing micrometric cavities, and integrated multi-wall carbon nanotubes (MWCNTs) into the scaffold. We combined microscopy, cell biology and calcium imaging, to investigate whether neonatal rat ventricular myocytes (NRVMs) cultured on the 3D-PDMS+MWCNT acquire a more viable and mature phenotype compared to control. We found that, when cultured in the 3D-PDMS+MWCNTs, NRVMs showed improved viability (p < 0.005 at day3) and more defined and mature sarcomeric phenotype compared to 3D PDMS control. These modifications were associated with an increase of connexin-43 gene expression, gap junction areas (p < 0.005 at day 3), and a more mature electrophysiological phenotype of syncytia and calcium transients. Finally, 3D-PDMS+MWCNT boosted NRVMs proliferation (p < 0.005 at day 3) while hindering cardiac fibroblasts proliferation compared to control PDMS. Thus, 3D-PDMS+MWCNT has the ability to promote viability, proliferation and functional maturation of cardiac myocytes. These properties are essential in cardiac tissue engineering and offer novel perspectives in the development of innovative therapies for cardiac repair.
- Published
- 2018
96. The global role of biotechnology for non communicable disorders
- Author
-
Mauro Giacca, Serena Zacchigna, Zacchigna, Serena, and Giacca, Mauro
- Subjects
0301 basic medicine ,Recombinant protein ,Small nuclei acids ,Non communicable disorders ,Developing country ,Bioengineering ,Context (language use) ,Applied Microbiology and Biotechnology ,Cell therapy ,World health ,03 medical and health sciences ,Non communicable disorder ,0302 clinical medicine ,Humans ,Diagnostic ,Noncommunicable Diseases ,Diagnostics ,Monoclonal antibodies ,Recombinant proteins ,Biotechnology ,Developing Countries ,Monoclonal antibodie ,business.industry ,General Medicine ,Small nuclei acid ,Early Diagnosis ,030104 developmental biology ,030220 oncology & carcinogenesis ,Business - Abstract
The World Health Organization (WHO) has tagged non-communicable diseases (NCDs) as one of the twenty-first century's major development challenges. NCDs account for over 15 million deaths annually and over 80% of those deaths occur in developing countries and among the poorest populations. Biotechnology presents unique opportunities to improve the early diagnosis and the treatment of NCDs. This review describes the major applications of biotechnology for a better clinical management of NCDs, i.e. the implementation of innovative diagnostic approaches and the production of innovative treatments, including those based on monoclonal antibodies, recombinant proteins, regulatory nucleic acids and cell-based therapies for regenerative medicine. In this context, it also examines the major challenges faced by biotechnology in developing countries.
- Published
- 2018
97. RNA interference therapeutics for cardiac regeneration
- Author
-
Hashim Ali, Josef Huntington, Manendra Pachauri, and Mauro Giacca
- Subjects
Stable nucleic acid lipid particle ,Myocardial Infarction ,Biology ,Viral vector ,Cardiac regeneration ,03 medical and health sciences ,0302 clinical medicine ,RNA interference ,microRNA ,Genetics ,medicine ,Animals ,Humans ,Regeneration ,Myocardial infarction ,030304 developmental biology ,0303 health sciences ,RNA ,medicine.disease ,3. Good health ,Cell biology ,MicroRNAs ,Heart failure ,RNA Interference ,030217 neurology & neurosurgery ,Developmental Biology - Abstract
There is an impelling need to develop new therapeutics for myocardial infarction and heart failure. A novel and exciting therapeutic possibility is to achieve cardiac regeneration through the stimulation of the endogenous capacity of cardiomyocytes to proliferate. Proof-of-concept evidence of microRNA-induced cardiac regeneration is available in both small and large animals using viral vectors. However, a clinically more applicable strategy is the development of lipid-mediated nanotechnologies for the administration of RNA therapeutics as synthetic molecules. The recent success of the Stable Nucleic Acid Lipid Particle (SNALP) platform for the generation of nanosized, efficient and non-inflammatory lipid nanoparticles paves the way to the development of injectable nanoformulations of microRNAs through cardiac catheterisation.
- Published
- 2021
98. Towards standardization of echocardiography for the evaluation of left ventricular function in adult rodents: a position paper of the ESC Working Group on Myocardial Function
- Author
-
Michele Ciccarelli, Evangelos P. Daskalopoulos, Luc Bertrand, Mauro Giacca, Florian Leuschner, Matteo Dal Ferro, Emilio Hirsch, Manuel Mayr, James Cimino, Michele Russo, Wolfgang A. Linke, Christophe Beauloye, André P. Lourenço, Antonio Cannatà, Lucie Carrier, Johannes Backs, Inês Falcão-Pires, Nazha Hamdani, Alessia Paldino, Pierluigi Lesizza, Florian Weinberger, Bruno Pinamonti, Serena Zacchigna, Dana Dawson, Ilona Cuijpers, Gianfranco Sinagra, Marco Oliveti, Diederik W. D. Kuster, Izhak Kehat, Stephane Heymans, Simone Vodret, Martine de Boer, Thomas Eschenhagen, Carlo G. Tocchetti, Alessandra Ghigo, Dirk J. Duncker, Jolanda van der Velden, Daniela Miranda-Silva, Thomas Thum, Zacchigna, Serena, Paldino, Alessia, Falcão-Pires, Inê, Daskalopoulos, Evangelos P, Dal Ferro, Matteo, Vodret, Simone, Lesizza, Pierluigi, Cannatà, Antonio, Miranda-Silva, Daniela, Lourenço, André P, Pinamonti, Bruno, Sinagra, Gianfranco, Weinberger, Florian, Eschenhagen, Thoma, Carrier, Lucie, Kehat, Izhak, Tocchetti, Carlo G, Russo, Michele, Ghigo, Alessandra, Cimino, Jame, Hirsch, Emilio, Dawson, Dana, Ciccarelli, Michele, Oliveti, Marco, Linke, Wolfgang A, Cuijpers, Ilona, Heymans, Stephane, Hamdani, Nazha, de Boer, Martine, Duncker, Dirk, Kuster, Diederik, van der Velden, Jolanda, Beauloye, Christophe, Bertrand, Luc, Mayr, Manuel, Giacca, Mauro, Leuschner, Florian, Backs, Johanne, Thum, Thomas, UCL - SSS/IREC/CARD - Pôle de recherche cardiovasculaire, and UCL - (SLuc) Service de pathologie cardiovasculaire
- Subjects
0301 basic medicine ,Cardiac function curve ,medicine.medical_specialty ,Biomedical Research ,Consensus ,Diastolic function ,Standardization ,systolic function ,Systole ,Physiology ,Diastole ,030204 cardiovascular system & hematology ,Working Group on Myocardial Function of the European Society of Cardiology ,Ventricular Function, Left ,Mice ,03 medical and health sciences ,0302 clinical medicine ,Physiology (medical) ,Internal medicine ,medicine ,Animals ,Animal model ,Pathological ,standardization ,Ventricular function ,business.industry ,Systolic function ,Echocardiography ,Animal models ,Myocardial function ,animal models ,Rats ,Disease Models, Animal ,030104 developmental biology ,Cardiovascular Diseases ,Cardiology ,Position paper ,Cardiology and Cardiovascular Medicine ,business - Abstract
Echocardiography is a reliable and reproducible method to assess non-invasively cardiac function in clinical and experimental research. Significant progress in the development of echocardiographic equipment and transducers has led to the successful translation of this methodology from humans to rodents, allowing for the scoring of disease severity and progression, testing of new drugs, and monitoring cardiac function in genetically modified or pharmacologically treated animals. However, as yet, there is no standardization in the procedure to acquire echocardiographic measurements in small animals. This position paper focuses on the appropriate acquisition and analysis of echocardiographic parameters in adult mice and rats, and provides reference values, representative images, and videos for the accurate and reproducible quantification of left ventricular function in healthy and pathological conditions. ispartof: CARDIOVASCULAR RESEARCH vol:117 issue:1 pages:43-59 ispartof: location:England status: published
- Published
- 2021
- Full Text
- View/download PDF
99. Calcium Signaling and Mitochondrial Function in Presenilin 2 Knock-Out Mice: Looking for Any Loss-of-Function Phenotype Related to Alzheimer’s Disease
- Author
-
Mauro Giacca, Alice Rossi, Luisa Galla, Valentina Giorgio, Chiara Gomiero, Lorena Zentilin, Tullio Pozzan, Tito Calì, Paola Pizzo, Elisa Greotti, Rossi, Alice, Galla, Luisa, Gomiero, Chiara, Zentilin, Lorena, Giacca, Mauro, Giorgio, Valentina, Calì, Tito, Pozzan, Tullio, Greotti, Elisa, and Pizzo, Paola
- Subjects
Amyloid beta ,Cell Respiration ,Citric Acid Cycle ,PS2–/– ,Mitochondrion ,Biology ,bioenergetics ,Endoplasmic Reticulum ,Presenilin ,Article ,Oxidative Phosphorylation ,03 medical and health sciences ,0302 clinical medicine ,presenilin 2 ,mitochondrial membrane potential ,Adenosine Triphosphate ,Cytosol ,Ca2+ signaling ,Alzheimer Disease ,PSEN2 ,Presenilin-2 ,PSEN1 ,Amyloid precursor protein ,Animals ,Calcium Signaling ,skin and connective tissue diseases ,lcsh:QH301-705.5 ,Loss function ,030304 developmental biology ,Calcium signaling ,Membrane Potential, Mitochondrial ,Mice, Knockout ,Neurons ,0303 health sciences ,bioenergetic ,General Medicine ,Cell biology ,neuronal hyperexcitability ,Mitochondria ,Mice, Inbred C57BL ,Alzheimer′s disease ,Phenotype ,lcsh:Biology (General) ,oxygen consumption rate ,biology.protein ,Glycolysis ,030217 neurology & neurosurgery - Abstract
Alzheimer&prime, s disease (AD) is the most common age-related neurodegenerative disorder in which learning, memory and cognitive functions decline progressively. Familial forms of AD (FAD) are caused by mutations in amyloid precursor protein (APP), presenilin 1 (PSEN1) and presenilin 2 (PSEN2) genes. Presenilin 1 (PS1) and its homologue, presenilin 2 (PS2), represent, alternatively, the catalytic core of the &gamma, secretase complex that, by cleaving APP, produces neurotoxic amyloid beta (A&beta, ) peptides responsible for one of the histopathological hallmarks in AD brains, the amyloid plaques. Recently, PSEN1 FAD mutations have been associated with a loss-of-function phenotype. To investigate whether this finding can also be extended to PSEN2 FAD mutations, we studied two processes known to be modulated by PS2 and altered by FAD mutations: Ca2+ signaling and mitochondrial function. By exploiting neurons derived from a PSEN2 knock-out (PS2&ndash, /&ndash, ) mouse model, we found that, upon IP3-generating stimulation, cytosolic Ca2+ handling is not altered, compared to wild-type cells, while mitochondrial Ca2+ uptake is strongly compromised. Accordingly, PS2&ndash, neurons show a marked reduction in endoplasmic reticulum&ndash, mitochondria apposition and a slight alteration in mitochondrial respiration, whereas mitochondrial membrane potential, and organelle morphology and number appear unchanged. Thus, although some alterations in mitochondrial function appear to be shared between PS2&ndash, and FAD-PS2-expressing neurons, the mechanisms leading to these defects are quite distinct between the two models. Taken together, our data appear to be difficult to reconcile with the proposal that FAD-PS2 mutants are loss-of-function, whereas the concept that PS2 plays a key role in sustaining mitochondrial function is here confirmed.
- Published
- 2021
100. Raptor is critical for increasing the mitochondrial proteome and skeletal muscle force during hypertrophy
- Author
-
Clara Türk, Frederik Telkamp, Achille Homère Tchampda Dondjang, Bert Blaauw, Alessia Geremia, Lorena Zentilin, Georgia Ana Dumitras, Marco Scalabrin, Mauro Giacca, Leonardo Nogara, Marcus Krüger, and Martina Baraldo
- Subjects
Male ,Proteome ,Knockout ,mTORC1 ,Mitochondrion ,Mechanistic Target of Rapamycin Complex 1 ,Inbred C57BL ,Biochemistry ,Muscle hypertrophy ,Mitochondrial Proteins ,Mice ,Mediator ,Genetics ,medicine ,Animals ,skeletal muscle ,Phosphorylation ,Muscle, Skeletal ,Molecular Biology ,Protein kinase B ,PI3K/AKT/mTOR pathway ,Mice, Knockout ,Chemistry ,rapamycin ,hypertrophy ,mitochondria ,mTOR ,Raptor ,Female ,Hypertrophy ,Mice, Inbred C57BL ,Mitochondria ,Regulatory-Associated Protein of mTOR ,Signal Transduction ,Skeletal muscle ,Skeletal ,medicine.disease ,Cell biology ,medicine.anatomical_structure ,Sarcopenia ,Muscle ,biological phenomena, cell phenomena, and immunity ,Biotechnology - Abstract
Loss of skeletal muscle mass and force is of critical importance in numerous pathologies, like age-related sarcopenia or cancer. It has been shown that the Akt-mTORC1 pathway is critical for stimulating adult muscle mass and function, however, it is unknown if mTORC1 is the only mediator downstream of Akt and which intracellular processes are required for functional muscle growth. Here, we show that loss of Raptor reduces muscle hypertrophy after Akt activation and completely prevents increases in muscle force. Interestingly, the residual hypertrophy after Raptor deletion can be completely prevented by administration of the mTORC1 inhibitor rapamycin. Using a quantitative proteomics approach we find that loss of Raptor affects the increases in mitochondrial proteins, while rapamycin mainly affects ribosomal proteins. Taken together, these results suggest that mTORC1 is the key mediator of Akt-dependent muscle growth and its regulation of the mitochondrial proteome is critical for increasing muscle force.
- Published
- 2021
Catalog
Discovery Service for Jio Institute Digital Library
For full access to our library's resources, please sign in.