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Article ; Online: Design and cloning of short hairpin RNAs (shRNAs) into a lentiviral silencing vector to study the function of selected proteins in neuronal apoptosis.

Canu, Nadia

Methods in molecular biology (Clifton, N.J.)

2015 Volume 1254, Page(s) 115–128

Abstract: Double-stranded RNA -mediated interference (RNAi ) is a new simple and fast research tool for shutting down genes and characterizes function of their respective proteins. Many strategies for design and delivery of siRNA to target cells are available. ... ...

Abstract	Double-stranded RNA -mediated interference (RNAi ) is a new simple and fast research tool for shutting down genes and characterizes function of their respective proteins. Many strategies for design and delivery of siRNA to target cells are available. Here, we describe the use of lentiviral short hairpin RNA (shRNA) RNA silencing to identify the involvement of d-serine racemase (SR )- an enzyme that syntheses d-serine to modulate glutamate- N-methyl-D-aspartate receptor- in regulating rat cerebellar granule neurons (CGN ) apoptosis. Apoptosis is induced by serum and KCl withdrawal and is detected with fluorometric caspase 3 assay.
MeSH term(s)	Animals ; Apoptosis/genetics ; Humans ; Lentivirus/genetics ; Molecular Biology/methods ; Neurons/metabolism ; RNA, Small Interfering/genetics ; Rats ; Receptors, N-Methyl-D-Aspartate/genetics
Chemical Substances	RNA, Small Interfering ; Receptors, N-Methyl-D-Aspartate
Language	English
Publishing date	2015
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ISSN	1940-6029
ISSN (online)	1940-6029
DOI	10.1007/978-1-4939-2152-2_9
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Article: NGF and the Amyloid Precursor Protein in Alzheimer's Disease: From Molecular Players to Neuronal Circuits.

Triaca, Viviana / Ruberti, Francesca / Canu, Nadia

Advances in experimental medicine and biology

2021 Volume 1331, Page(s) 145–165

Abstract: Alzheimer's disease (AD), one of the most common causes of dementia in elderly people, is characterized by progressive impairment in cognitive function, early degeneration of basal forebrain cholinergic neurons (BFCNs), abnormal metabolism of the amyloid ...

Abstract	Alzheimer's disease (AD), one of the most common causes of dementia in elderly people, is characterized by progressive impairment in cognitive function, early degeneration of basal forebrain cholinergic neurons (BFCNs), abnormal metabolism of the amyloid precursor protein (APP), amyloid beta-peptide (Aβ) depositions, and neurofibrillary tangles. According to the cholinergic hypothesis, dysfunction of acetylcholine-containing neurons in the basal forebrain contributes markedly to the cognitive decline observed in AD. In addition, the neurotrophic factor hypothesis posits that the loss nerve growth factor (NGF) signalling in AD may account for the vulnerability to atrophy of BFCNs and consequent impairment of cholinergic functions. Though acetylcholinesterase inhibitors provide only partial and symptomatic relief to AD patients, emerging data from in vivo magnetic resonance imaging (MRI) and positron emission tomography (PET) studies in mild cognitive impairment (MCI) and AD patients highlight the early involvement of BFCNs in MCI and the early phase of AD. These data support the cholinergic and neurotrophic hypotheses of AD and suggest new targets for AD therapy.Different mechanisms account for selective vulnerability of BFCNs to AD pathology, with regard to altered metabolism of APP and tau. In this review, we provide a general overview of the current knowledge of NGF and APP interplay, focusing on the role of APP in regulating NGF receptors trafficking/signalling and on the involvement of NGF in modulating phosphorylation of APP, which in turn controls APP intracellular trafficking and processing. Moreover, we highlight the consequences of APP interaction with p75NTR and TrkA receptor, which share the same binding site within the APP juxta-membrane domain. We underline the importance of insulin dysmetabolism in AD pathology, in the light of our recent data showing that overlapping intracellular signalling pathways stimulated by NGF or insulin can be compensatory. In particular, NGF-based signalling is able to ameliorates deficiencies in insulin signalling in the medial septum of 3×Tg-AD mice. Finally, we present an overview of NGF-regulated microRNAs (miRNAs). These small non-coding RNAs are involved in post-transcriptional regulation of gene expression , and we focus on a subset that are specifically deregulated in AD and thus potentially contribute to its pathology.
MeSH term(s)	Aged ; Alzheimer Disease ; Amyloid beta-Peptides ; Amyloid beta-Protein Precursor/genetics ; Animals ; Humans ; Mice ; Nerve Growth Factor ; Neurons
Chemical Substances	Amyloid beta-Peptides ; Amyloid beta-Protein Precursor ; Nerve Growth Factor (9061-61-4)
Language	English
Publishing date	2021-08-28
Publishing country	United States
Document type	Journal Article ; Review
ISSN	2214-8019 ; 0065-2598
ISSN (online)	2214-8019
ISSN	0065-2598
DOI	10.1007/978-3-030-74046-7_10
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Article ; Online: New insights on the influence of free d-aspartate metabolism in the mammalian brain during prenatal and postnatal life.

Errico, Francesco / Cuomo, Mariella / Canu, Nadia / Caputo, Viviana / Usiello, Alessandro

Biochimica et biophysica acta. Proteins and proteomics

2020 Volume 1868, Issue 10, Page(s) 140471

Abstract: Free d-aspartate is abundant in the mammalian embryonic brain. However, following the postnatal onset of the catabolic d-aspartate oxidase (DDO) activity, cerebral d-aspartate levels drastically decrease, remaining constantly low throughout life. d- ... ...

Abstract	Free d-aspartate is abundant in the mammalian embryonic brain. However, following the postnatal onset of the catabolic d-aspartate oxidase (DDO) activity, cerebral d-aspartate levels drastically decrease, remaining constantly low throughout life. d-Aspartate stimulates both glutamatergic NMDA receptors (NMDARs) and metabotropic Glu5 receptors. In rodents, short-term d-aspartate exposure increases spine density and synaptic plasticity, and improves cognition. Conversely, persistently high d-Asp levels produce NMDAR-dependent neurotoxic effects, leading to precocious neuroinflammation and cell death. These pieces of evidence highlight the dichotomous impact of d-aspartate signaling on NMDAR-dependent processes and, in turn, unveil a neuroprotective role for DDO in preventing the detrimental effects of excessive d-aspartate stimulation during aging. Here, we will focus on the in vivo influence of altered d-aspartate metabolism on the modulation of glutamatergic functions and its involvement in translational studies. Finally, preliminary data on the role of embryonic d-aspartate in the mouse brain will also be reviewed.
MeSH term(s)	Age Factors ; Aging/genetics ; Aging/metabolism ; Animals ; Biomarkers ; Brain/anatomy & histology ; Brain/growth & development ; Brain/metabolism ; D-Aspartate Oxidase/genetics ; D-Aspartate Oxidase/metabolism ; D-Aspartic Acid/metabolism ; Dietary Supplements ; Disease Susceptibility ; Female ; Gene Expression Regulation, Developmental ; Gene Expression Regulation, Enzymologic ; Mammals/metabolism ; Memory ; Neurogenesis ; Neuroprotection ; Pregnancy ; Receptors, N-Methyl-D-Aspartate/genetics ; Receptors, N-Methyl-D-Aspartate/metabolism
Chemical Substances	Biomarkers ; Receptors, N-Methyl-D-Aspartate ; D-Aspartic Acid (4SR0Q8YD1X) ; D-Aspartate Oxidase (EC 1.4.3.1)
Language	English
Publishing date	2020-06-17
Publishing country	Netherlands
Document type	Journal Article ; Research Support, Non-U.S. Gov't ; Review
ZDB-ID	2918798-9
ISSN	1878-1454 ; 1570-9639
ISSN (online)	1878-1454
ISSN	1570-9639
DOI	10.1016/j.bbapap.2020.140471
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Article ; Online: Fyn Tyrosine Kinase Elicits Amyloid Precursor Protein Tyr682 Phosphorylation in Neurons from Alzheimer's Disease Patients.

Iannuzzi, Filomena / Sirabella, Rossana / Canu, Nadia / Maier, Thorsten J / Annunziato, Lucio / Matrone, Carmela

Cells

2020 Volume 9, Issue 8

Abstract: Alzheimer's disease (AD) is an incurable neurodegenerative disorder with a few early detection strategies. We previously proposed the amyloid precursor protein (APP) tyrosine 682 (Tyr682) residue as a valuable target for the development of new innovative ...

Abstract	Alzheimer's disease (AD) is an incurable neurodegenerative disorder with a few early detection strategies. We previously proposed the amyloid precursor protein (APP) tyrosine 682 (Tyr682) residue as a valuable target for the development of new innovative pharmacologic or diagnostic interventions in AD. Indeed, when APP is phosphorylated at Tyr682, it is forced into acidic neuronal compartments where it is processed to generate neurotoxic amyloid β peptides. Of interest, Fyn tyrosine kinase (TK) interaction with APP Tyr682 residue increases in AD neurons. Here we proved that when Fyn TK was overexpressed it elicited APP Tyr682 phosphorylation in neurons from healthy donors and promoted the amyloidogenic APP processing with Aβ peptides accumulation and neuronal death. Phosphorylation of APP at Tyr (pAPP-Tyr) increased in neurons of AD patients and AD neurons that exhibited high pAPP-Tyr also had higher Fyn TK activity. Fyn TK inhibition abolished the pAPP-Tyr and reduced Aβ42 secretion in AD neurons. In addition, the multidomain adaptor protein Fe65 controlled the Fyn-mediated pAPP-Tyr, warranting the possibility of targeting the Fe65-APP-Fyn pathway to develop innovative strategies in AD. Altogether, these results strongly emphasize the relevance of focusing on pAPP Tyr682 either for diagnostic purposes, as an early biomarker of the disease, or for pharmacological targeting, using Fyn TKI.
MeSH term(s)	Adult ; Aged ; Aged, 80 and over ; Alzheimer Disease/metabolism ; Alzheimer Disease/pathology ; Amyloid beta-Protein Precursor/genetics ; Amyloid beta-Protein Precursor/metabolism ; Biomarkers/metabolism ; Cells, Cultured ; Female ; Humans ; Induced Pluripotent Stem Cells/metabolism ; Male ; Middle Aged ; Neural Stem Cells/metabolism ; Neurons/metabolism ; Phosphorylation/drug effects ; Phosphorylation/genetics ; Protein Kinase Inhibitors/pharmacology ; Proto-Oncogene Proteins c-fyn/antagonists & inhibitors ; Proto-Oncogene Proteins c-fyn/genetics ; Proto-Oncogene Proteins c-fyn/metabolism ; Transfection ; Tyrosine/metabolism
Chemical Substances	APP protein, human ; Amyloid beta-Protein Precursor ; Biomarkers ; Protein Kinase Inhibitors ; Tyrosine (42HK56048U) ; FYN protein, human (EC 2.7.10.2) ; Proto-Oncogene Proteins c-fyn (EC 2.7.10.2)
Language	English
Publishing date	2020-07-30
Publishing country	Switzerland
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2661518-6
ISSN	2073-4409 ; 2073-4409
ISSN (online)	2073-4409
ISSN	2073-4409
DOI	10.3390/cells9081807
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Article ; Online: Fyn Tyrosine Kinase Elicits Amyloid Precursor Protein Tyr682 Phosphorylation in Neurons from Alzheimer’s Disease Patients

Filomena Iannuzzi / Rossana Sirabella / Nadia Canu / Thorsten J Maier / Lucio Annunziato / Carmela Matrone

Cells, Vol 9, Iss 1807, p

2020 Volume 1807

Abstract: Alzheimer’s disease (AD) is an incurable neurodegenerative disorder with a few early detection strategies. We previously proposed the amyloid precursor protein (APP) tyrosine 682 (Tyr682) residue as a valuable target for the development of new innovative ...

Abstract	Alzheimer’s disease (AD) is an incurable neurodegenerative disorder with a few early detection strategies. We previously proposed the amyloid precursor protein (APP) tyrosine 682 (Tyr682) residue as a valuable target for the development of new innovative pharmacologic or diagnostic interventions in AD. Indeed, when APP is phosphorylated at Tyr682, it is forced into acidic neuronal compartments where it is processed to generate neurotoxic amyloid β peptides. Of interest, Fyn tyrosine kinase (TK) interaction with APP Tyr682 residue increases in AD neurons. Here we proved that when Fyn TK was overexpressed it elicited APP Tyr682 phosphorylation in neurons from healthy donors and promoted the amyloidogenic APP processing with Aβ peptides accumulation and neuronal death. Phosphorylation of APP at Tyr (pAPP-Tyr) increased in neurons of AD patients and AD neurons that exhibited high pAPP-Tyr also had higher Fyn TK activity. Fyn TK inhibition abolished the pAPP-Tyr and reduced Aβ42 secretion in AD neurons. In addition, the multidomain adaptor protein Fe65 controlled the Fyn-mediated pAPP-Tyr, warranting the possibility of targeting the Fe65-APP-Fyn pathway to develop innovative strategies in AD. Altogether, these results strongly emphasize the relevance of focusing on pAPP Tyr682 either for diagnostic purposes, as an early biomarker of the disease, or for pharmacological targeting, using Fyn TKI.
Keywords	amyloid precursor protein ; amyloid beta ; Fyn tyrosine kinase ; Tyr682 residue ; YENPTY domain ; Biology (General) ; QH301-705.5
Subject code	616
Language	English
Publishing date	2020-07-01T00:00:00Z
Publisher	MDPI AG
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article: New insights on the influence of free d-aspartate metabolism in the mammalian brain during prenatal and postnatal life

Errico, Francesco / Cuomo, Mariella / Canu, Nadia / Caputo, Viviana / Usiello, Alessandro

Biochimica et biophysica acta. 2020 Oct., v. 1868, no. 10

2020

Abstract	Free d-aspartate is abundant in the mammalian embryonic brain. However, following the postnatal onset of the catabolic d-aspartate oxidase (DDO) activity, cerebral d-aspartate levels drastically decrease, remaining constantly low throughout life. d-Aspartate stimulates both glutamatergic NMDA receptors (NMDARs) and metabotropic Glu5 receptors. In rodents, short-term d-aspartate exposure increases spine density and synaptic plasticity, and improves cognition. Conversely, persistently high d-Asp levels produce NMDAR-dependent neurotoxic effects, leading to precocious neuroinflammation and cell death. These pieces of evidence highlight the dichotomous impact of d-aspartate signaling on NMDAR-dependent processes and, in turn, unveil a neuroprotective role for DDO in preventing the detrimental effects of excessive d-aspartate stimulation during aging. Here, we will focus on the in vivo influence of altered d-aspartate metabolism on the modulation of glutamatergic functions and its involvement in translational studies. Finally, preliminary data on the role of embryonic d-aspartate in the mouse brain will also be reviewed.
Keywords	D-aspartate oxidase ; brain ; cell death ; cognition ; metabolism ; mice ; neuroplasticity ; neuroprotective effect ; neurotoxicity
Language	English
Dates of publication	2020-10
Publishing place	Elsevier B.V.
Document type	Article
Note	NAL-AP-2-clean
ZDB-ID	2918798-9
ISSN	1878-1454 ; 1570-9639
ISSN (online)	1878-1454
ISSN	1570-9639
DOI	10.1016/j.bbapap.2020.140471
Database	NAL-Catalogue (AGRICOLA)

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Article ; Online: Insulin and serine metabolism as sex-specific hallmarks of Alzheimer's disease in the human hippocampus.

Maffioli, Elisa / Murtas, Giulia / Rabattoni, Valentina / Badone, Beatrice / Tripodi, Farida / Iannuzzi, Filomena / Licastro, Danilo / Nonnis, Simona / Rinaldi, Anna Maria / Motta, Zoraide / Sacchi, Silvia / Canu, Nadia / Tedeschi, Gabriella / Coccetti, Paola / Pollegioni, Loredano

Cell reports

2022 Volume 40, Issue 10, Page(s) 111271

Abstract: Healthy aging is an ambitious aspiration for humans, but neurodegenerative disorders, such as Alzheimer's disease (AD), strongly affect quality of life. Using an integrated omics approach, we investigate alterations in the molecular composition of ... ...

Abstract	Healthy aging is an ambitious aspiration for humans, but neurodegenerative disorders, such as Alzheimer's disease (AD), strongly affect quality of life. Using an integrated omics approach, we investigate alterations in the molecular composition of postmortem hippocampus samples of healthy persons and individuals with AD. Profound differences are apparent between control and AD male and female cohorts in terms of up- and downregulated metabolic pathways. A decrease in the insulin response is evident in AD when comparing the female with the male group. The serine metabolism (linked to the glycolytic pathway and generating the N-methyl-D-aspartate [NMDA] receptor coagonist D-serine) is also significantly modulated: the D-Ser/total serine ratio represents a way to counteract age-related cognitive decline in healthy men and during AD onset in women. These results show how AD changes and, in certain respects, almost reverses sex-specific proteomic and metabolomic profiles, highlighting how different pathophysiological mechanisms are active in men and women.
MeSH term(s)	Alzheimer Disease/metabolism ; Female ; Hippocampus/metabolism ; Humans ; Insulin/metabolism ; Male ; Proteomics ; Quality of Life ; Receptors, N-Methyl-D-Aspartate/metabolism ; Serine/metabolism
Chemical Substances	Insulin ; Receptors, N-Methyl-D-Aspartate ; Serine (452VLY9402)
Language	English
Publishing date	2022-07-29
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2649101-1
ISSN	2211-1247 ; 2211-1247
ISSN (online)	2211-1247
ISSN	2211-1247
DOI	10.1016/j.celrep.2022.111271
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Article ; Online: Research on occupational diseases in the absence of occupational data: a mixed-method study among cancer registries of Western Switzerland.

Plys, Ekaterina / Bovio, Nicolas / Arveux, Patrick / Bergeron, Yvan / Bulliard, Jean-Luc / Elia, Nadia / Fournier, Evelyne / Konzelmann, Isabelle / Maspoli, Manuela / Rapiti Aylward, Elisabetta / Guseva Canu, Irina

Swiss medical weekly

2022 Volume 152, Page(s) w30127

Abstract: The contribution of occupation-related diseases to the global burden of disease is greatly underestimated, mainly due to a shortage of occupational exposure data. This problem is particularly salient in Switzerland, where no estimates of occupation- ... ...

Abstract	The contribution of occupation-related diseases to the global burden of disease is greatly underestimated, mainly due to a shortage of occupational exposure data. This problem is particularly salient in Switzerland, where no estimates of occupation-related disease burden exist, even for the well-recognised occupational cancers, such as malignant pleural mesothelioma and lung cancer. To overcome this situation, we launched a research project "Examining Cancers and Labour Indicators to assess the Burden" (ExCaLIBur). Within this project, we aimed to assess the need for and quality (i.e., completeness, accuracy and precision) of occupation registration in all cancer registries of Western Switzerland. We also aimed to find a relevant and feasible strategy to collect this information in the future. We applied a mixed research method. We observed that, independently of the level of precision (5-3-2-1-digit aggregation level), the accuracy was lesser in the registries that were able to actively search and verify occupational information. Overall, the distinction of occupations based on the 3-digit code presents an acceptable compromise in terms of precision. Having such occupations registered in all, or most, Swiss cancer registries routinely would obviously be valuable for epidemiological surveillance of occupational cancers in Switzerland. However, it seems less obvious how these data could fulfill the research objectives, since a better precision than 3-digit occupational coding is challenging to achieve. Currently, the collection of occupational data by the Swiss cancer registries remains feasible in the frame of specific research projects on occupational cancers. However, available data sources, as well as lack of financial and human resources, will continue to affect quality of the collected occupation data. Therefore, the usage of the standardised questionnaire retracing the individual occupational history to enable further assessment of individual exposure to potential occupational hazards is recommended. However, this approach will disable the Swiss registries to insuring their epidemiological surveillance mission with respect to occupational cancers in Switzerland, for which national statistics remain limited.
MeSH term(s)	Humans ; Lung Neoplasms/epidemiology ; Occupational Diseases/epidemiology ; Occupational Exposure ; Registries ; Switzerland/epidemiology
Language	English
Publishing date	2022-02-22
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2036179-8
ISSN	1424-3997 ; 1424-7860
ISSN (online)	1424-3997
ISSN	1424-7860
DOI	10.4414/smw.2022.w30127
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Article ; Online: A standardized approach to empirically define reliable assignment thresholds and appropriate management categories in deeply introgressed populations.

Caniglia, Romolo / Galaverni, Marco / Velli, Edoardo / Mattucci, Federica / Canu, Antonio / Apollonio, Marco / Mucci, Nadia / Scandura, Massimo / Fabbri, Elena

Scientific reports

2020 Volume 10, Issue 1, Page(s) 2862

Abstract: Anthropogenic hybridization is recognized as a major threat to the long-term survival of natural populations. While identifying F1 hybrids might be simple, the detection of older admixed individuals is far from trivial and it is still debated whether ... ...

Abstract	Anthropogenic hybridization is recognized as a major threat to the long-term survival of natural populations. While identifying F1 hybrids might be simple, the detection of older admixed individuals is far from trivial and it is still debated whether they should be targets of management. Examples of anthropogenic hybridization have been described between wolves and domestic dogs, with numerous cases detected in the Italian wolf population. After selecting appropriate wild and domestic reference populations, we used empirical and simulated 39-autosomal microsatellite genotypes, Bayesian assignment and performance analyses to develop a workflow to detect different levels of wolf x dog admixture. Membership proportions to the wild cluster (q
MeSH term(s)	Animals ; Animals, Wild/genetics ; Bayes Theorem ; Conservation of Natural Resources ; Dogs ; Genetic Variation/genetics ; Genetics, Population ; Genotype ; Hybridization, Genetic/genetics ; Microsatellite Repeats/genetics ; Polymorphism, Single Nucleotide/genetics ; Wolves/genetics
Language	English
Publishing date	2020-02-18
Publishing country	England
Document type	Journal Article
ZDB-ID	2615211-3
ISSN	2045-2322 ; 2045-2322
ISSN (online)	2045-2322
ISSN	2045-2322
DOI	10.1038/s41598-020-59521-2
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Article ; Online: In vivo macrophage engineering reshapes the tumor microenvironment leading to eradication of liver metastases.

Kerzel, Thomas / Giacca, Giovanna / Beretta, Stefano / Bresesti, Chiara / Notaro, Marco / Scotti, Giulia Maria / Balestrieri, Chiara / Canu, Tamara / Redegalli, Miriam / Pedica, Federica / Genua, Marco / Ostuni, Renato / Kajaste-Rudnitski, Anna / Oshima, Masanobu / Tonon, Giovanni / Merelli, Ivan / Aldrighetti, Luca / Dellabona, Paolo / Coltella, Nadia /

Doglioni, Claudio / Rancoita, Paola M V / Sanvito, Francesca / Naldini, Luigi / Squadrito, Mario Leonardo

Cancer cell

2023 Volume 41, Issue 11, Page(s) 1892–1910.e10

Abstract: Liver metastases are associated with poor response to current pharmacological treatments, including immunotherapy. We describe a lentiviral vector (LV) platform to selectively engineer liver macrophages, including Kupffer cells and tumor-associated ... ...

Abstract	Liver metastases are associated with poor response to current pharmacological treatments, including immunotherapy. We describe a lentiviral vector (LV) platform to selectively engineer liver macrophages, including Kupffer cells and tumor-associated macrophages (TAMs), to deliver type I interferon (IFNα) to liver metastases. Gene-based IFNα delivery delays the growth of colorectal and pancreatic ductal adenocarcinoma liver metastases in mice. Response to IFNα is associated with TAM immune activation, enhanced MHC-II-restricted antigen presentation and reduced exhaustion of CD8
MeSH term(s)	Humans ; Mice ; Animals ; CD8-Positive T-Lymphocytes ; CTLA-4 Antigen/metabolism ; Tumor Microenvironment/genetics ; Macrophages ; Liver Neoplasms/genetics ; Liver Neoplasms/therapy ; Liver Neoplasms/pathology
Chemical Substances	CTLA-4 Antigen
Language	English
Publishing date	2023-10-19
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2078448-X
ISSN	1878-3686 ; 1535-6108
ISSN (online)	1878-3686
ISSN	1535-6108
DOI	10.1016/j.ccell.2023.09.014
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