4.4 Review

Two-pore channels: going with the flows

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Acidic Ca2+ stores and immune-cell function

Lianne C. Davis et al.

Summary: Acidic organelles serve as intracellular Ca2+ stores and play important roles in immune-cell function, impacting processes such as membrane trafficking, vesicle fusion/fission, and secretion. Different endo-lysosomal Ca2+ channels on the same vesicles can affect various downstream physiological functions.

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Summary: TRPML1 is a Ca2+-permeable, nonselective cation channel expressed in endolysosomes, which is activated by cooperation between PI(3,5)P2 and rapamycin. The cryoEM structures and electrophysiology studies provide insights into the activation mechanism of TRPML1.

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SARM1 is a multi-functional NAD(P)ase with prominent base exchange activity, all regulated by multiple physiologically relevant NAD metabolites

Carlo Angeletti et al.

Summary: SARM1 is an important enzyme involved in programmed axon death. It has multiple activities including the highest base exchange activity at neutral pH. The base exchange also occurs in neurons and may be a source of the calcium-mobilizing agent NaADP. The regulation of SARM1 by free pyridines, NADP, and nicotinic acid riboside has therapeutic implications.

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Article Cell Biology

Lysosomal TPCN (two pore segment channel) inhibition ameliorates beta-amyloid pathology and mitigates memory impairment in Alzheimer disease

Benjamin Chun-Kit Tong et al.

Summary: This study identified a mechanism of impairment in the macroautophagy/autophagy-lysosomal pathway (ALP) in Alzheimer's disease, mediated by the lysosomal TPCN2/TPC2 channel. Targeting TPCN2 was shown to restore lysosomal function and reduce amyloid plaque accumulation, suggesting a potential therapeutic strategy for AD treatment in the future.

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Cheng-Chang Chen et al.

Summary: There is growing evidence that ion channels in the endolysosomal system play a crucial role in the pathology of infectious diseases and cancer. This review focuses on the characteristics and functions of endolysosomal cation channels and discusses their importance as potential drug targets.

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Neurophysiological functions and pharmacological tools of acidic and non-acidic Ca2+ stores

Lora L. Martucci et al.

Summary: Ca2+ signaling plays a crucial role in various cell functions in the brain. Acidic Ca2+ stores in different regions of neurons provide local sources of Ca2+ to regulate synaptic transmission and other functions. This review discusses the recruitment mechanism of acidic Ca2+ stores and focuses on the role of endolysosomal two-pore channels (TPCs) and their physiological agonist, as well as their interaction with other receptors. Additionally, this review introduces new pharmacological tools and animal mutant models for studying acidic Ca2+ stores in brain function and dysfunction.

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Localized TPC1-mediated Ca2+ release from endolysosomes contributes to myoseptal junction development in zebrafish

Keira L. Rice et al.

Summary: The endolysosomal ion channel TPC1 plays a crucial role in the formation and maintenance of myoseptal junctions (MJs) in developing zebrafish embryos. It generates localized Ca2+ signals that are essential for capturing and attaching slow skeletal muscle cells at MJs. TPC1-decorated endolysosomes are dynamically associated with MJs through a microtubule-dependent mechanism and regulate endolysosomal trafficking and transmembrane protein distribution.

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Cardiac glycosides stimulate endocytosis of GLUT1 via intracellular Na+,K+-ATPase alpha 3-isoform in human cancer cells

Takuto Fujii et al.

Summary: CGs can inhibit glucose metabolism of cancer cells by inhibiting Na+,K+-ATPase and decreasing the expression of GLUT1. The binding of CGs with intracellular alpha 3NaK elicits NAADP-mediated calcium mobilization.

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Proton-gated anion transport governs macropinosome shrinkage

Mariia Zeziulia et al.

Summary: Zeziulia et al. identified the proton-activated Cl- channel ASOR/TMEM206 as necessary for the shrinkage of macrophage macropinosomes, which plays a crucial role in sorting and trafficking of cellular organelles. The activation of ASOR requires depolarization mediated by Na+ and acidification by transporters such as H+-ATPases.

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NAADP-binding proteins find their identity

Jonathan S. Marchant et al.

Summary: Nicotinic acid adenine dinucleotide phosphate (NAADP) is a second messenger that releases Ca2+ from endosomes and lysosomes by activating two-pore channels (TPCs). Two different NAADP-binding proteins (NAADP-BPs), JPT2 and LSM12, have recently been identified, bridging the gap between NAADP generation and NAADP activation of TPCs. The discovery of these NAADP-BPs will facilitate future studies on the molecular mechanism of NAADP action.

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Nicotinic Acid Adenine Dinucleotide Phosphate Induces Intracellular Ca2+ Signalling and Stimulates Proliferation in Human Cardiac Mesenchymal Stromal Cells

Pawan Faris et al.

Summary: Nicotinic acid adenine dinucleotide phosphate (NAADP) regulates Ca2+ release in lysosomes through dual pore channels 1 and 2, playing an important role in cardiac mesenchymal stromal cells (C-MSCs). These signals are amplified by the Ca2+-induced Ca2+ release mechanism, affecting cell proliferation and signaling.

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Nicotinic acid adenine dinucleotide phosphate activates two-pore channel TPC1 to mediate lysosomal Ca2+release in endothelial colony-forming cells

Francesco Moccia et al.

Summary: The study demonstrates that NAADP-induced TPC1-mediated Ca(2+)release can selectively regulate endothelial functions in circulating ECFCs by mobilizing lysosomal Ca(2+)store.

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TPC2 promotes choroidal angiogenesis and inflammation in a mouse model of neovascular age-related macular degeneration

Yanfen Li et al.

Summary: Age-related macular degeneration (AMD) is a common cause of blindness among the elderly, classified as dry or neovascular. Research shows that the endolysosomal two-pore cation channel TPC2 plays a key role in neovascularization and immune activation in AMD, and blocking TPC2 can reduce related factors levels in the pathological process of AMD.

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Nicola Clementi et al.

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UCSF ChimeraX: Structure visualization for researchers, educators, and developers

Eric F. Pettersen et al.

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Human genome diversity data reveal that L564P is the predominant TPC2 variant and a prerequisite for the blond hair associated M484L gain-of-function effect

Julia Bock et al.

Summary: The study provides a comprehensive analysis of the role of the endo-lysosomal cation channel TPC2 in various diseases and traits, including metabolism, obesity, diabetes, infectious diseases, cancer, and pigmentation defects. By identifying a predominant TPC2 variant L564P globally and characterizing its effects on channel activity, along with discovering novel gain-of-function variants, the research sheds light on the ethnic and geographical distribution of TPC2 polymorphisms. Additionally, the study highlights the potential of TPC2 as a pharmacological drug target due to the identified variations in different human populations.

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Oxidation of Protein Kinase A Regulatory Subunit PKARIα Protects Against Myocardial Ischemia-Reperfusion Injury by Inhibiting Lysosomal-Triggered Calcium Release

Jillian N. Simon et al.

Summary: The study revealed that PKARI alpha disulfide formation is significantly increased in response to I/R in both humans and mice, leading to enhanced AKAP binding and preferential localization to the lysosome. This redox-dependent regulation of lysosomal channels by PKARI alpha plays a critical role in protecting the heart from extensive injury and could serve as a novel target for cardioprotective therapeutics.

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Dual NADPH oxidases DUOX1 and DUOX2 synthesize NAADP and are necessary for Ca2+ signaling during T cell activation

Feng Gu et al.

Summary: The formation of Ca2+ microdomains during T cell activation is initiated by the production of nicotinic acid adenine dinucleotide phosphate (NAADP) from its reduced form NAADPH. NADPH oxidases NOX and DUOX play crucial roles in this process, with DUOX2 and G6PD catalyzing a redox cycle that rapidly produces and degrades NAADP through NAADPH.

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Two-pore channels affect EGF receptor signaling by receptor trafficking and expression

Thomas Mueller et al.

Summary: Two-pore channels (TPCs) play a crucial role in regulating calcium ion currents between endosomes and lysosomes, affecting membrane fusion and fission events as well as intracellular trafficking. The study suggests that TPCs not only regulate the EGFR transportation network, but also influence EGFR signaling and expression.

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HN1L/JPT2: A signaling protein that connects NAADP generation to Ca2+ microdomain formation

Hannes G. Roggenkamp et al.

Summary: This study identified a new NAADP-binding protein, HN1L/JPT2, which connects NAADP formation with the activation of RYR channels, facilitating Ca2+ release. This protein plays a crucial role in the earliest phase of T cell activation.

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Essential requirement for JPT2 in NAADP-evoked Ca2+ signaling

Gihan S. Gunaratne et al.

Summary: The study identified JPT2 as an essential component of the NAADP receptor complex, required for TPC-dependent Ca2+ signaling and control of coronaviral entry through the endolysosomal system.

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Flavonoids increase melanin production and reduce proliferation, migration and invasion of melanoma cells by blocking endolysosomal/melanosomal TPC2

Ponsawan Netcharoensirisuk et al.

Summary: Studies have shown an inverse correlation between TPC2 activity in endolysosomes and melanosomes and melanin production, melanoma proliferation, migration, and invasion. Inhibiting TPC2 can increase melanin synthesis in melanosomes by interfering with tyrosinase activity and decrease melanoma progression driven by MITF through GSK3 beta-mediated MITF degradation.

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NAADP-induced intracellular calcium ion is mediated by the TPCs (two-pore channels) in hypoxia-induced pulmonary arterial hypertension

Wen Hu et al.

Summary: The study investigated the pathogenesis of pulmonary arterial hypertension in rat models and found that two-pore segment channels TPC1/2 play a role in the disease, potentially serving as a therapeutic target.

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Lsm12 is an NAADP receptor and a two-pore channel regulatory protein required for calcium mobilization from acidic organelles

Jiyuan Zhang et al.

Summary: Researchers identified Lsm12 as an NAADP receptor that mediates Ca2+ release from lysosomes via NAADP binding on its Lsm domain. Lsm12 co-localizes with TPC2 and plays a crucial role in NAADP signaling.

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Glucose and NAADP trigger elementary intracellular β-cell Ca2+ signals

Paula Maria Heister et al.

Summary: Insulin release from pancreatic beta-cells is associated with highly localized intracellular Ca2+ signals, which may be influenced by global Ca2+ signals and SERCA-based sequestration mechanisms. Previous work has demonstrated the importance of NAADP-induced Ca2+ release in stimulus-secretion coupling in beta-cells.

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The key role of the central cavity in sodium transport through ligand-gated two-pore channels

Stefan Milenkovic et al.

Summary: This article discusses the important role of mammalian two-pore channels in maintaining cellular life processes, particularly the significance of hTPC2 in viral infections. Through molecular simulations, a model for sodium ion transport is proposed.

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Gene editing and synthetically accessible inhibitors reveal role for TPC2 in HCC cell proliferation and tumor growth

Martin Mueller et al.

Summary: Our study unveils the role of TPC2 in cancer and demonstrates that TPC2 knockout reduces cancer cell proliferation and affects energy metabolism. We have also developed simplified analogs of TPC2 inhibitors with stronger antiproliferative properties against cancer cells.

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Anand Saminathan et al.

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Resting state structure of the hyperdepolarization activated two-pore channel 3

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Contribution of NAADP to Glutamate-Evoked Changes in Ca2+Homeostasis in Mouse Hippocampal Neurons

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Genetic Inactivation of Two-Pore Channel 1 Impairs Spatial Learning and Memory

Robert Theodor Mallmann et al.

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TPC1 deficiency or blockade augments systemic anaphylaxis and mast cell activity

Elisabeth Arlt et al.

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David L. Prole et al.

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Rapamycin directly activates lysosomal mucolipin TRP channels independent of mTOR

Xiaoli Zhang et al.

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Einar K. Krogsaeter et al.

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Mining of Ebola virus entry inhibitors identifies approved drugs as two-pore channel pore blockers

Christopher J. Penny et al.

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Phosphatidylinositol-3,5-bisphosphate lipid-binding-induced activation of the human two-pore channel 2

Sonja A. Kirsch et al.

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Two-pore channels mediated receptor-operated Ca2+ entry in pulmonary artery smooth muscle cells in response to hypoxia

Yongliang Jiang et al.

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mTORC1 controls lysosomal Ca2+ release through the two-pore channel TPC2

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mTORC1 controls lysosomal Ca2+ release through the two-pore channel TPC2

Oluseye A. Ogunbayo et al.

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Hippocampal mGluR1-dependent long-term potentiation requires NAADP-mediated acidic store Ca2+ signaling

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Glutamate induces autophagy via the two-pore channels in neural cells

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Liposomes as a Putative Tool to Investigate NAADP Signaling in Vasculogenesis

Francesca Di Nezza et al.

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TPC2 polymorphisms associated with a hair pigmentation phenotype in humans result in gain of channel function by independent mechanisms

Yu-Kai Chao et al.

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The human two-pore channel 1 is modulated by cytosolic and luminal calcium

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Naringenin Impairs Two-Pore Channel 2 Activity And Inhibits VEGF-Induced Angiogenesis

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Ong Nam Phuong Nguyen et al.

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Metabolic alterations derived from absence of Two-Pore Channel 1 at cardiac level

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Ca2+/H+ exchange by acidic organelles regulates cell migration in vivo

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Structure, inhibition and regulation of two-pore channel TPC1 from Arabidopsis thaliana

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TPC2 controls pigmentation by regulating melanosome pH and size

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A melanosomal two-pore sodium channel regulates pigmentation

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NAADP-Dependent Ca2+ Signaling Controls Melanoma Progression, Metastatic Dissemination and Neoangiogenesis

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Expression of Ca2+-permeable two-pore channels rescues NAADP signalling in TPC-deficient cells

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Two-Pore Channel 2 activity is required for slow muscle cell-generated Ca2+ signaling during myogenesis in intact zebrafish

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TPC2 mediates new mechanisms of platelet dense granule membrane dynamics through regulation of Ca2+ release

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Lysosomal Two-pore Channel Subtype 2 (TPC2) Regulates Skeletal Muscle Autophagic Signaling

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Dysregulation of lysosomal morphology by pathogenic LRRK2 is corrected by TPC2 inhibition

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Anthony J. Morgan et al.

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TPC1 Has Two Variant Isoforms, and Their Removal Has Different Effects on Endo-Lysosomal Functions Compared to Loss of TPC2

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The voltage-gated sodium channel TPC1 confers endolysosomal excitability

Chunlei Cang et al.

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The Two-pore channel (TPC) interactome unmasks isoform-specific roles for TPCs in endolysosomal morphology and cell pigmentation

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Reconstituted Human TPC1 Is a Proton-Permeable Ion Channel and Is Activated by NAADP or Ca2+

Samantha J. Pitt et al.

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High susceptibility to fatty liver disease in two-pore channel 2-deficient mice

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mTOR Regulates Lysosomal ATP-Sensitive Two-Pore Na+ Channels to Adapt to Metabolic State

Chunlei Cang et al.

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Bidirectional Ca2+ signaling occurs between the endoplasmic reticulum and acidic organelles

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Joseph A. Mindell

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Triggering of Ca2+ signals by NAADP-gated two-pore channels: a role for membrane contact sites?

Sandip Patel et al.

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NAADP Activates Two-Pore Channels on T Cell Cytolytic Granules to Stimulate Exocytosis and Killing

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Leucine-rich repeat kinase 2 regulates autophagy through a calcium-dependent pathway involving NAADP

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In vivo, Pikfyve generates PI(3,5)P2, which serves as both a signaling lipid and the major precursor for PI5P

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Characterization of Two-pore Channel 2 (TPCN2)-mediated Ca2+ Currents in Isolated Lysosomes

Michael Schieder et al.

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A cation counterflux supports lysosomal acidification

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Recruitment of NAADP-sensitive acidic Ca2+ stores by glutamate

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Identification of a chemical probe for NAADP by virtual screening

Edmund Naylor et al.

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NAADP-mediated Ca2+ signaling via type 1 ryanodine receptor in T cells revealed by a synthetic NAADP antagonist

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NAADP induces pH changes in the lumen of acidic Ca2+ stores

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