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Article: Targeting phosphoinositide signaling in cancer: relevant techniques to study lipids and novel avenues for therapeutic intervention.

Llorente, Alicia / Loughran, Ryan M / Emerling, Brooke M

Frontiers in cell and developmental biology

2023 Volume 11, Page(s) 1297355

Abstract: Phosphoinositides serve as essential players in numerous biological activities and are critical for overall cellular function. Due to their complex chemical structures, localization, and low abundance, current challenges in the phosphoinositide field ... ...

Abstract	Phosphoinositides serve as essential players in numerous biological activities and are critical for overall cellular function. Due to their complex chemical structures, localization, and low abundance, current challenges in the phosphoinositide field include the accurate measurement and identification of specific variants, particularly those with acyl chains. Researchers are intensively developing innovative techniques and approaches to address these challenges and advance our understanding of the impact of phosphoinositide signaling on cellular biology. This article provides an overview of recent advances in the study of phosphoinositides, including mass spectrometry, lipid biosensors, and real-time activity assays using fluorometric sensors. These methodologies have proven instrumental for a comprehensive exploration of the cellular distribution and dynamics of phosphoinositides and have shed light on the growing significance of these lipids in human health and various pathological processes, including cancer. To illustrate the importance of phosphoinositide signaling in disease, this perspective also highlights the role of a family of lipid kinases named phosphatidylinositol 5-phosphate 4-kinases (PI5P4Ks), which have recently emerged as exciting therapeutic targets for cancer treatment. The ongoing exploration of phosphoinositide signaling not only deepens our understanding of cellular biology but also holds promise for novel interventions in cancer therapy.
Language	English
Publishing date	2023-10-25
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2737824-X
ISSN	2296-634X
ISSN	2296-634X
DOI	10.3389/fcell.2023.1297355
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Article ; Online: Lipids in Extracellular Vesicles: What Can Be Learned about Membrane Structure and Function?

Skotland, Tore / Llorente, Alicia / Sandvig, Kirsten

Cold Spring Harbor perspectives in biology

2023 Volume 15, Issue 8

Abstract: Extracellular vesicles, such as exosomes, can be used as interesting models to study the structure and function of biological membranes as these vesicles contain only one membrane (i.e., one lipid bilayer). In addition to lipids, they contain proteins, ... ...

Abstract	Extracellular vesicles, such as exosomes, can be used as interesting models to study the structure and function of biological membranes as these vesicles contain only one membrane (i.e., one lipid bilayer). In addition to lipids, they contain proteins, nucleic acids, and various other molecules. The lipid composition of exosomes is here compared to HIV particles and detergent-resistant membranes, which also have a high content of sphingolipids, cholesterol, and phosphatidylserine (PS). We discuss interactions between the lipids in the two bilayers, and especially those between PS 18:0/18:1 in the inner leaflet and the very-long-chain sphingolipids in the outer leaflet, and the importance of cholesterol for these interactions. We also briefly discuss the involvement of ether-linked phospholipids (PLs) in such lipid raft-like structures, and the possible involvement of these and other lipid classes in the formation of exosomes. The urgent need to improve the quality of quantitative lipidomic studies is highlighted.
MeSH term(s)	Lipid Bilayers/chemistry ; Lipid Bilayers/metabolism ; Cell Membrane/metabolism ; Sphingolipids/metabolism ; Extracellular Vesicles/metabolism ; Cholesterol
Chemical Substances	Lipid Bilayers ; Sphingolipids ; Cholesterol (97C5T2UQ7J)
Language	English
Publishing date	2023-08-01
Publishing country	United States
Document type	Journal Article ; Review ; Research Support, Non-U.S. Gov't
ISSN	1943-0264
ISSN (online)	1943-0264
DOI	10.1101/cshperspect.a041415
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Article: Lipid Biomarkers in Liquid Biopsies: Novel Opportunities for Cancer Diagnosis.

Sagini, Krizia / Urbanelli, Lorena / Buratta, Sandra / Emiliani, Carla / Llorente, Alicia

Pharmaceutics

2023 Volume 15, Issue 2

Abstract: Altered cellular metabolism is a well-established hallmark of cancer. Although most studies have focused on the metabolism of glucose and glutamine, the upregulation of lipid metabolism is also frequent in cells undergoing oncogenic transformation. In ... ...

Abstract	Altered cellular metabolism is a well-established hallmark of cancer. Although most studies have focused on the metabolism of glucose and glutamine, the upregulation of lipid metabolism is also frequent in cells undergoing oncogenic transformation. In fact, cancer cells need to meet the enhanced demand of plasma membrane synthesis and energy production to support their proliferation. Moreover, lipids are precursors of signaling molecules, termed lipid mediators, which play a role in shaping the tumor microenvironment. Recent methodological advances in lipid analysis have prompted studies aimed at investigating the whole lipid content of a sample (lipidome) to unravel the complexity of lipid changes in cancer patient biofluids. This review focuses on the application of mass spectrometry-based lipidomics for the discovery of cancer biomarkers. Here, we have summarized the main lipid alteration in cancer patients' biofluids and uncovered their potential use for the early detection of the disease and treatment selection. We also discuss the advantages of using biofluid-derived extracellular vesicles as a platform for lipid biomarker discovery. These vesicles have a molecular signature that is a fingerprint of their originating cells. Hence, the analysis of their molecular cargo has emerged as a promising strategy for the identification of sensitive and specific biomarkers compared to the analysis of the unprocessed biofluid.
Language	English
Publishing date	2023-01-28
Publishing country	Switzerland
Document type	Journal Article ; Review
ZDB-ID	2527217-2
ISSN	1999-4923
ISSN	1999-4923
DOI	10.3390/pharmaceutics15020437
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Article ; Online: Biodistribution, pharmacokinetics and excretion studies of intravenously injected nanoparticles and extracellular vesicles: Possibilities and challenges.

Skotland, Tore / Iversen, Tore Geir / Llorente, Alicia / Sandvig, Kirsten

Advanced drug delivery reviews

2022 Volume 186, Page(s) 114326

Abstract: There is a large interest in developing nanoparticles and extracellular vesicles for delivery of therapeutics or imaging agents. Regulatory approval of such products requires knowledge about their biodistribution, metabolism and excretion. We here ... ...

Abstract	There is a large interest in developing nanoparticles and extracellular vesicles for delivery of therapeutics or imaging agents. Regulatory approval of such products requires knowledge about their biodistribution, metabolism and excretion. We here discuss possibilities and challenges of methods used for such studies, which most often are performed after labelling with radioactive isotopes or fluorescent molecules. It is important to evaluate if the labelled and unlabeled products can be expected to behave similarly in the body. Furthermore, one needs to critically consider whether the labels are still associated with the product at the time of analyses. We discuss advantages and disadvantages of different imaging modalities such as PET, SPECT, MRI, CT, ultrasound and optical imaging for whole-body biodistribution, and describe how to estimate the amount of labelled product in harvested organs and tissue. Microscopy of cells and tissues and various mass spectrometry methods are also discussed in this review.
MeSH term(s)	Extracellular Vesicles ; Humans ; Nanoparticles/chemistry ; Optical Imaging ; Tissue Distribution ; Tomography, Emission-Computed, Single-Photon
Language	English
Publishing date	2022-05-16
Publishing country	Netherlands
Document type	Journal Article ; Review ; Research Support, Non-U.S. Gov't
ZDB-ID	639113-8
ISSN	1872-8294 ; 0169-409X
ISSN (online)	1872-8294
ISSN	0169-409X
DOI	10.1016/j.addr.2022.114326
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Article ; Online: PIP kinases: A versatile family that demands further therapeutic attention.

Llorente, Alicia / Arora, Gurpreet K / Grenier, Shea F / Emerling, Brooke M

Advances in biological regulation

2022 Volume 87, Page(s) 100939

MeSH term(s)	Humans ; Phosphotransferases ; Signal Transduction ; Phosphatidylinositol 4,5-Diphosphate
Chemical Substances	Phosphotransferases (EC 2.7.-) ; Phosphatidylinositol 4,5-Diphosphate
Language	English
Publishing date	2022-12-05
Publishing country	England
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	2667413-0
ISSN	2212-4934 ; 2212-4926
ISSN (online)	2212-4934
ISSN	2212-4926
DOI	10.1016/j.jbior.2022.100939
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Article ; Online: An emerging focus on lipids in extracellular vesicles.

Skotland, Tore / Sagini, Krizia / Sandvig, Kirsten / Llorente, Alicia

Advanced drug delivery reviews

2020 Volume 159, Page(s) 308–321

Abstract: Extracellular vesicles contain a lipid bilayer membrane that protects the encapsulated material, such as proteins, nucleic acids, lipids and metabolites, from the extracellular environment. These vesicles are released from cells via different mechanisms. ...

Abstract	Extracellular vesicles contain a lipid bilayer membrane that protects the encapsulated material, such as proteins, nucleic acids, lipids and metabolites, from the extracellular environment. These vesicles are released from cells via different mechanisms. During recent years extracellular vesicles have been studied as possible biomarkers for different diseases, as biological nanoparticles for drug delivery, and in basic studies as a tool to understand the structure of biological membranes and the mechanisms involved in vesicular trafficking. Lipids are essential molecular components of extracellular vesicles, but at the moment our knowledge about the lipid composition and the function of lipids in these vesicles is limited. However, the interest of the research community in these molecules is increasing as their role in extracellular vesicles is starting to be acknowledged. In this review, we will present the status of the field and describe what is needed to bring it forward.
MeSH term(s)	Animals ; Extracellular Vesicles/metabolism ; Humans ; Lipid Metabolism ; Lipids
Chemical Substances	Lipids
Language	English
Publishing date	2020-03-07
Publishing country	Netherlands
Document type	Journal Article ; Research Support, Non-U.S. Gov't ; Review
ZDB-ID	639113-8
ISSN	1872-8294 ; 0169-409X
ISSN (online)	1872-8294
ISSN	0169-409X
DOI	10.1016/j.addr.2020.03.002
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Article ; Online: Ebulin l Is Internalized in Cells by Both Clathrin-Dependent and -Independent Mechanisms and Does Not Require Clathrin or Dynamin for Intoxication.

Iglesias, Rosario / Ferreras, José M / Llorente, Alicia / Citores, Lucía

Toxins

2021 Volume 13, Issue 2

Abstract: Ebulin l is an A-B toxin, and despite the presence of a B chain, this toxin displays much less toxicity to cells than the potent A-B toxin ricin. Here, we studied the binding, mechanisms of endocytosis, and intracellular pathway followed by ebulin l and ... ...

Abstract	Ebulin l is an A-B toxin, and despite the presence of a B chain, this toxin displays much less toxicity to cells than the potent A-B toxin ricin. Here, we studied the binding, mechanisms of endocytosis, and intracellular pathway followed by ebulin l and compared it with ricin. COS-1 cells and HeLa cells with inducible synthesis of a mutant dynamin (K44A) were used in this study. The transport of these toxins was measured using radioactively or fluorescently labeled toxins. The data show that ebulin l binds to cells to a lesser extent than ricin. Moreover, the expression of mutant dynamin does not affect the endocytosis, degradation, or toxicity of ebulin l. However, the inhibition of clathrin-coated pit formation by acidification of the cytosol reduced ebulin l endocytosis but not toxicity. Remarkably, unlike ricin, ebulin l is not transported through the Golgi apparatus to intoxicate the cells and ebulin l induces apoptosis as the predominant cell death mechanism. Therefore, after binding to cells, ebulin l is taken up by clathrin-dependent and -independent endocytosis into the endosomal/lysosomal system, but there is no apparent role for clathrin and dynamin in productive intracellular routing leading to intoxication.
MeSH term(s)	Animals ; Apoptosis/drug effects ; COS Cells ; Chlorocebus aethiops ; Clathrin/metabolism ; Clathrin-Coated Vesicles/metabolism ; Dynamins/genetics ; Dynamins/metabolism ; Endocytosis ; HeLa Cells ; Humans ; Mutation ; Protein Transport ; Proteolysis ; Ribosome Inactivating Proteins, Type 2/metabolism ; Ribosome Inactivating Proteins, Type 2/toxicity ; Ricin/metabolism
Chemical Substances	Clathrin ; Ribosome Inactivating Proteins, Type 2 ; ebulin 1 protein, Sambucus ebulus ; Ricin (9009-86-3) ; Dynamins (EC 3.6.5.5)
Language	English
Publishing date	2021-01-30
Publishing country	Switzerland
Document type	Comparative Study ; Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2518395-3
ISSN	2072-6651 ; 2072-6651
ISSN (online)	2072-6651
ISSN	2072-6651
DOI	10.3390/toxins13020102
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Article ; Online: Potential of miRNAs in urinary extracellular vesicles for management of active surveillance in prostate cancer patients.

Ramirez-Garrastacho, Manuel / Berge, Viktor / Linē, Aija / Llorente, Alicia

British journal of cancer

2021 Volume 126, Issue 3, Page(s) 492–501

Abstract: Background: Active surveillance is an alternative to radical treatment for patients with low-risk prostate cancer, which could also benefit some patients with intermediate risk. We have investigated the use of miRNA in urinary extracellular vesicles to ... ...

Abstract	Background: Active surveillance is an alternative to radical treatment for patients with low-risk prostate cancer, which could also benefit some patients with intermediate risk. We have investigated the use of miRNA in urinary extracellular vesicles to stratify these patients. Methods: NGS was performed to profile the miRNAs from small urinary extracellular vesicles in a cohort of 70 patients with prostate cancer ISUP Grade 1, 2 or 3. The most promising candidates were then analysed by RT-qPCR in a new cohort of 60 patients. Results: NGS analysis identified nine miRNAs differentially expressed in at least one of the comparisons. The largest differences were found with miR-1290 (Grade 3 vs. 1), miR-320a-3p (Grade 3 vs. 2) and miR-155-5p (Grade 2 vs. 1). Combinations of 2-3 miRNAs were able to differentiate between two ISUP grades with an AUC 0.79-0.88. RT-qPCR analysis showed a similar trend for miR-186-5p and miR-30e-5p to separate Grade 3 from 2, and miR-320a-3p to separate Grade 2 from 1. Conclusions: Using NGS, we have identified several miRNAs that discriminate between prostate cancer patients with ISUP Grades 1, 2 and 3. Moreover, miR-186-5p, miR-320a-3p and miR-30e-5p showed a similar behaviour in an independent cohort using an alternative analytical method. Our results show that miRNAs from urinary vesicles can be potentially useful as liquid biopsies for active surveillance.
MeSH term(s)	Biomarkers, Tumor/genetics ; Biomarkers, Tumor/urine ; Extracellular Vesicles/genetics ; Extracellular Vesicles/pathology ; Gene Expression Regulation, Neoplastic ; Humans ; Male ; MicroRNAs/genetics ; MicroRNAs/urine ; Neoplasm Grading ; Prostatectomy/methods ; Prostatic Neoplasms/genetics ; Prostatic Neoplasms/pathology ; Prostatic Neoplasms/surgery ; Prostatic Neoplasms/urine ; ROC Curve ; Watchful Waiting/methods
Chemical Substances	Biomarkers, Tumor ; MicroRNAs
Language	English
Publishing date	2021-11-22
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	80075-2
ISSN	1532-1827 ; 0007-0920
ISSN (online)	1532-1827
ISSN	0007-0920
DOI	10.1038/s41416-021-01598-1
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Article ; Online: Extracellular Vesicles as Novel Players in Kidney Disease.

Blijdorp, Charles J / Burger, Dylan / Llorente, Alicia / Martens-Uzunova, Elena S / Erdbrügger, Uta

Journal of the American Society of Nephrology : JASN

2022 Volume 33, Issue 3, Page(s) 467–471

MeSH term(s)	Biomarkers ; Exosomes ; Extracellular Vesicles ; Female ; Humans ; Kidney Diseases ; Male
Chemical Substances	Biomarkers
Language	English
Publishing date	2022-02-07
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	1085942-1
ISSN	1533-3450 ; 1046-6673
ISSN (online)	1533-3450
ISSN	1046-6673
DOI	10.1681/ASN.2021091232
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Article ; Online: The expanding organelle lipidomes: current knowledge and challenges.

Sarmento, Maria J / Llorente, Alicia / Petan, Toni / Khnykin, Denis / Popa, Iuliana / Nikolac Perkovic, Matea / Konjevod, Marcela / Jaganjac, Morana

Cellular and molecular life sciences : CMLS

2023 Volume 80, Issue 8, Page(s) 237

Abstract: Lipids in cell membranes and subcellular compartments play essential roles in numerous cellular processes, such as energy production, cell signaling and inflammation. A specific organelle lipidome is characterized by lipid synthesis and metabolism, ... ...

Abstract	Lipids in cell membranes and subcellular compartments play essential roles in numerous cellular processes, such as energy production, cell signaling and inflammation. A specific organelle lipidome is characterized by lipid synthesis and metabolism, intracellular trafficking, and lipid homeostasis in the organelle. Over the years, considerable effort has been directed to the identification of the lipid fingerprints of cellular organelles. However, these fingerprints are not fully characterized due to the large variety and structural complexity of lipids and the great variability in the abundance of different lipid species. The process becomes even more challenging when considering that the lipidome differs in health and disease contexts. This review summarizes the information available on the lipid composition of mammalian cell organelles, particularly the lipidome of the nucleus, mitochondrion, endoplasmic reticulum, Golgi apparatus, plasma membrane and organelles in the endocytic pathway. The lipid compositions of extracellular vesicles and lamellar bodies are also described. In addition, several examples of subcellular lipidome dynamics under physiological and pathological conditions are presented. Finally, challenges in mapping organelle lipidomes are discussed.
MeSH term(s)	Animals ; Lipids/analysis ; Lipidomics ; Lipid Metabolism ; Organelles/metabolism ; Cell Nucleus/metabolism ; Mitochondria/metabolism ; Mammals
Chemical Substances	Lipids
Language	English
Publishing date	2023-08-02
Publishing country	Switzerland
Document type	Journal Article ; Review
ZDB-ID	1358415-7
ISSN	1420-9071 ; 1420-682X
ISSN (online)	1420-9071
ISSN	1420-682X
DOI	10.1007/s00018-023-04889-3
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