Development of ROS-triggered masked liposomes for activated cellular delivery using a charge balance strategy.

We report a charge balance strategy for reactive oxygen species (ROS)-triggered activation of liposome cell delivery by unmasking cationic membranes. Zeta potential experiments, microplate-based vesicle interaction assays, and cellular delivery studies confirmed that modification of anionic lipid 1 by ROS led to the uncaging of cationic liposomes, thereby driving cellular association.

Subcellular imaging of lipids and sugars using genetically encoded proximity sensors.

Live cell imaging of lipids and other metabolites is a long-standing challenge in cell biology. Bioorthogonal labeling tools allow for the conjugation of fluorophores to several phospholipid classes, but cannot discern their trafficking between adjacent organelles or asymmetry across individual membrane leaflets. Here we present fluorogen-activating coincidence sensing (FACES), a chemogenetic tool capable of quantitatively imaging subcellular lipid pools and reporting their transbilayer orientation in living cells.

Probing Glycerolipid Metabolism using a Caged Clickable Glycerol-3-Phosphate Probe.

In this study, we present the probe SATE-G3P-N as a novel tool for metabolic labeling of glycerolipids (GLs) to investigate lipid metabolism in yeast cells. By introducing a clickable azide handle onto the glycerol backbone, this probe enables general labeling of glycerolipids. Additionally, this probe contains a caged phosphate moiety at the glycerol sn-3 position to not only facilitate probe uptake by masking negative charge but also to bypass the phosphorylation step crucial for initiating phospholipid synthesis, thereby enhancing phospholipid labeling.

The small molecule CBR-5884 inhibits the phosphatidylserine synthase.

Unlabelled: Systemic infections by spp. are associated with high mortality rates, partly due to limitations in current antifungals, highlighting the need for novel drugs and drug targets. The fungal phosphatidylserine synthase, Cho1, from is a logical antifungal drug target due to its importance in virulence, absence in the host, and conservation among fungal pathogens.

Nuclear phosphoinositide signaling promotes YAP/TAZ-TEAD transcriptional activity in breast cancer.

The Hippo pathway effectors Yes-associated protein 1 (YAP) and its homolog TAZ are transcriptional coactivators that control gene expression by binding to TEA domain (TEAD) family transcription factors. The YAP/TAZ-TEAD complex is a key regulator of cancer-specific transcriptional programs, which promote tumor progression in diverse types of cancer, including breast cancer. Despite intensive efforts, the YAP/TAZ-TEAD complex in cancer has remained largely undruggable due to an incomplete mechanistic understanding.

Development of a bis-pyrene phospholipid probe for fluorometric detection of phospholipase A inhibition.

In this work, we report the design, synthesis, and application of a bis-pyrene phospholipid probe for detection of phospholipase A action through changes in pyrene monomer and excimer fluorescence intensities. Continuous fluorometric assays enabled detection of the activities of multiple PLA enzymes as well as the decrease in catalysis by PLA from honey bee venom caused by the inhibitor p-bromo phenacylbromide. Thin-layer chromatography and mass spectrometry analysis were also used to validate probe hydrolysis by PLA.

Harnessing Clickable Acylated Glycerol Probes as Chemical Tools for Tracking Glycerolipid Metabolism.

We report the use of clickable monoacylglycerol (MAG) analogs as probes for the labeling of glycerolipids during lipid metabolism. Incorporation of azide tags onto the glycerol region was pursued to develop probes that would label glycerolipids, in which the click tag would not be removed through processes including acyl chain and headgroup remodeling. Analysis of clickable MAG probes containing acyl chains of different length resulted in widely variable cell imaging and cytotoxicity profiles.

Development of GTP-responsive liposomes by exchanging the metal-DPA binding site in a synthetic lipid switch.

We report stimuli-responsive liposomes that selectively release encapsulated contents upon treatment with guanosine triphosphate (GTP) over a wide variety of phosphorylated metabolites, validated by fluorescence-based leakage assays. Significant changes in liposome self-assembly properties were also observed. Our results showcase the potential of this platform for triggered release applications.

Copper-responsive liposomes for triggered cargo release employing a picolinamide-lipid conjugate.

In this work, we report triggered content release from liposomes brought about by copper chelation to a synthetic lipid switch containing a picolinamide headgroup. Fluorescence-based dye-leakage assays showcase release of carboxyfluorescein dye cargo upon copper treatment and control of liposomal release based on copper abundance. Our results additionally show that this platform is selective for copper and is accompanied by significant changes to liposome properties upon treatment with this ion.

Metabolite-Responsive Liposomes Employing Synthetic Lipid Switches Driven by Molecular Recognition Principles.

The ability to exert control over lipid properties, including structure, charge, function, and self-assembly characteristics is a powerful tool that can be implemented to achieve a wide range of biomedical applications. Examples in this arena include the development of caged lipids for controlled activation of signaling properties, metabolic labeling strategies for tracking lipid biosynthesis, lipid activity probes for identifying cognate binding partners, approaches for membrane assembly, and liposome triggered release strategies. In this Account, we describe recent advancements in the latter area entailing the development of stimuli-responsive liposomes through programmable changes to lipid self-assembly properties, which can be harnessed to drive the release of encapsulated contents toward applications including drug delivery.

Sticking the Landing: Enhancing Liposomal Cell Delivery using Reversible Covalent Chemistry and Caged Targeting Groups.

Liposomes are highly effective nanocarriers for encapsulating and delivering a wide range of therapeutic cargo. While advancements in liposome design have improved several pharmacological characteristics, an important area that would benefit from further progress involves cellular targeting and entry. In this concept article, we will focus on recent progress utilizing strategies including reversible covalent bonding and caging groups to activate liposomal cell entry.

Bis-Boronic Acid Liposomes for Carbohydrate Recognition and Cellular Delivery.

Liposomes are effective therapeutic nanocarriers due to their ability to encapsulate and enhance the pharmacokinetic properties of a wide range of drugs and diagnostic agents. A primary area in which improvement is needed for liposomal drug delivery is to maximize the delivery of these nanocarriers to cells. Cell membrane glycans provide exciting targets for liposomal delivery since they are often densely clustered on cell membranes and glycan overabundance and aberrant glycosylation patterns are a common feature of diseased cells.

Cyclic Disulfide Liposomes for Membrane Functionalization and Cellular Delivery.

Liposomes are effective therapeutic delivery nanocarriers due to their ability to encapsulate and enhance the pharmacokinetic properties of a wide range of therapeutics. Two primary areas in which improvement is needed for liposomal drug delivery is to enhance the ability to infiltrate cells and to facilitate derivatization of the liposome surface. Herein, we report a liposome platform incorporating a cyclic disulfide lipid (CDL) for the dual purpose of enhancing cell entry and functionalizing the liposome membrane through thiol-disulfide exchange.

Reactive Oxygen Species (ROS) Activated Liposomal Cell Delivery using a Boronate-Caged Guanidine Lipid.

We report boronate-caged guanidine-lipid 1 that activates liposomes for cellular delivery only upon uncaging of this compound by reactive oxygen species (ROS) to produce cationic lipid products. These liposomes are designed to mimic the exceptional cell delivery properties of cell-penetrating peptides (CPPs), while the inclusion of the boronate cage is designed to enhance selectivity such that cell entry will only be activated in the presence of ROS. Boronate uncaging by hydrogen peroxide was verified by mass spectrometry and zeta potential (ZP) measurements.

Liposome triggered content release through molecular recognition of inositol trisphosphate.

A stimuli-responsive liposomal platform that is selectively activated by inositol 1,4,5-trisphosphate (IP) over eleven other phosphorylated metabolites is reported. Dye release assays validated dose-dependent release of both hydrophilic and hydrophobic cargo driven by IP, showcasing the potential of this platform for triggered release and sensing applications.

ATP-Responsive Liposomes via Screening of Lipid Switches Designed to Undergo Conformational Changes upon Binding Phosphorylated Metabolites.

Liposomal delivery vehicles can dramatically enhance drug transport. However, their clinical application requires enhanced control over content release at diseased sites. For this reason, triggered release strategies have been explored, although a limited toolbox of stimuli has thus far been developed.

Atomic view of an amyloid dodecamer exhibiting selective cellular toxic vulnerability in acute brain slices.

Atomic structures of amyloid oligomers that capture the neurodegenerative disease pathology are essential to understand disease-state causes and finding cures. Here we investigate the G6W mutation of the cytotoxic, hexameric amyloid model KV11. The mutation results into an asymmetric dodecamer composed of a pair of 30° twisted antiparallel β-sheets.

Zinc Triggered Release of Encapsulated Cargo from Liposomes via a Synthetic Lipid Switch.

Liposomes are effective nanocarriers due to their ability to encapsulate and deliver a wide variety of therapeutics. However, therapeutic potential would be improved by enhanced control over the release of drug cargo. Zinc ions provide exciting new targets for stimuli-responsive lipid design due to their overly abundant concentrations associated with diseased cells.

Demolish and Rebuild: Controlling Lipid Self-Assembly toward Triggered Release and Artificial Cells.

The ability to modulate the structures of lipid membranes, predicated on our nuanced understanding of the properties that drive and alter lipid self-assembly, has opened up many exciting biological applications. In this Perspective, we focus on two endeavors in which the same principles are invoked to achieve completely opposite results. On one hand, controlled liposome decomposition enables triggered release of encapsulated cargo through the development of synthetic lipid switches that perturb lipid packing in the presence of disease-associated stimuli.

Strategies for altering lipid self-assembly to trigger liposome cargo release.

While liposomes have proven to be effective drug delivery nanocarriers, their therapeutic attributes could be improved through the development of clinically viable triggered release strategies in which encapsulated drug contents could be selectively released at the sites of diseased cells. As such, a significant amount of research has been reported involving the development of stimuli-responsive liposomes and a broad range of strategies have been explored for driving content release. These have included the introduction of trigger groups at either the lipid headgroup or within the acyl chains that alter lipid self-assembly properties of known lipids as well as the rational design of lipid analogs programed to undergo conformational changes induced by events such as binding interactions.

Reactive Oxygen Species-Responsive Liposomes via Boronate-Caged Phosphatidylethanolamine.

Liposomes have proven to be effective nanocarriers due to their ability to encapsulate and deliver a wide variety of therapeutic cargo. A key goal of liposome research is to enhance control over content release at diseased sites. Though a number of stimuli have been explored for triggering liposomal release, reactive oxygen species (ROS), which have received significantly less attention, provide excellent targets due to their key roles in biology and overabundance in diseased cells.

Calcium-responsive liposomes: Toward ion-mediated targeted drug delivery.

Liposomes are clinically approved supramolecular drug delivery platforms due to their ability to enhance the pharmacokinetic properties of encapsulated therapeutic agents. A key point for advancing liposomal drug delivery would be to control the timing and location of cargo release to maximize drug potency and minimize side effects. Toward this end, triggered release approaches have been developed that exploit either pathophysiological stimuli (passive release) including pH or external stimuli (active release) such as light.

A General Approach to Enzyme-Responsive Liposomes.

Liposomes are effective nanocarriers due to their ability to deliver encapsulated drugs to diseased cells. Nevertheless, liposome delivery would be improved by enhancing the ability to control the release of contents at the target site. While various stimuli have been explored for triggering liposome release, enzymes provide excellent targets due to their common overexpression in diseased cells.

Lipid Switches: Stimuli-Responsive Liposomes through Conformational Isomerism Driven by Molecular Recognition.

Advancements in the field of liposomal drug carriers have culminated in greatly improved delivery properties. An important aspect of this work entails development of designer liposomes for release of contents triggered by environmental changes. The majority of these systems are driven by chemical reactions in the presence of different stimuli.

Boronic acid liposomes for cellular delivery and content release driven by carbohydrate binding.

Boronic acid liposomes enable triggered content release and cell delivery driven by carbohydrate binding. Dye release assays using hydrophilic and hydrophobic fluorophores validate dose-dependent release upon carbohydrate treatment. Microscopy results indicate dramatic enhancements in cell delivery, showcasing the prospects of boronic acid lipids for drug delivery.