Publications by authors named "Wanqing Du"

15 Publications

  • Page 1 of 1

Nuclear translocation of the 4-pass transmembrane protein Tspan8.

Cell Res 2021 Jun 7. Epub 2021 Jun 7.

State Key Laboratory of Oncogenes and Related Genes, Department of Oncology, Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.

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http://dx.doi.org/10.1038/s41422-021-00522-9DOI Listing
June 2021

Studying Mitochondrial Network Formation by In Vivo and In Vitro Reconstitution Assay.

Methods Mol Biol 2021 ;2276:333-341

Institute for Biomedical Materials & Devices (IBMD), Faculty of Science, University of Technology Sydney, Sydney, NSW, Australia.

Mitochondria change their morphologies from small isolated vesicles to large continuous networks across the cell cycles. The mitochondrial network formation (MNF) plays an important role in maintaining mitochondrial DNA integrity and interchanging mitochondrial materials. The disruption of the mitochondrial network affects mitochondrial functions, such as ATP production, integration of metabolism, calcium homeostasis, and regulation of apoptosis, leading to the abnormal development and several human diseases including neurodegenerative disease. In this unit, we describe the method of studying MNF, which is driven by microtubule-dependent motor protein, by in vivo imaging and single-molecule in vitro reconstitution assays.
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http://dx.doi.org/10.1007/978-1-0716-1266-8_25DOI Listing
January 2021

Mitocytosis, a migrasome-mediated mitochondrial quality-control process.

Cell 2021 May;184(11):2896-2910.e13

State Key Laboratory of Membrane Biology, Tsinghua University-Peking University Joint Centre for Life Sciences, Beijing Frontier Research Center for Biological Structure, School of Life Sciences, Tsinghua University, Beijing 100084, China. Electronic address:

Damaged mitochondria need to be cleared to maintain the quality of the mitochondrial pool. Here, we report mitocytosis, a migrasome-mediated mitochondrial quality-control process. We found that, upon exposure to mild mitochondrial stresses, damaged mitochondria are transported into migrasomes and subsequently disposed of from migrating cells. Mechanistically, mitocytosis requires positioning of damaged mitochondria at the cell periphery, which occurs because damaged mitochondria avoid binding to inward motor proteins. Functionally, mitocytosis plays an important role in maintaining mitochondrial quality. Enhanced mitocytosis protects cells from mitochondrial stressor-induced loss of mitochondrial membrane potential (MMP) and mitochondrial respiration; conversely, blocking mitocytosis causes loss of MMP and mitochondrial respiration under normal conditions. Physiologically, we demonstrate that mitocytosis is required for maintaining MMP and viability in neutrophils in vivo. We propose that mitocytosis is an important mitochondrial quality-control process in migrating cells, which couples mitochondrial homeostasis with cell migration.
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http://dx.doi.org/10.1016/j.cell.2021.04.027DOI Listing
May 2021

Acupuncture for Hashimoto thyroiditis: study protocol for a randomized controlled trial.

Trials 2021 Jan 21;22(1):74. Epub 2021 Jan 21.

The Department of Acupuncture, Dongzhimen Hospital, Beijing University of Chinese Medicine, Beijing, 100700, China.

Background: The incidence rate of Hashimoto thyroiditis (HT) has gradually increased in recent years. There has been no specific etiological treatment for HT. Even though with normal level of thyroid hormone, the patients may still suffer from various clinical symptoms, such as anterior neck discomfort, fatigue, and mood swings, which seriously impair their quality of life. Acupuncture has long been used in the treatment of thyroid diseases, but there has been no related standardized clinical study as of today. This study aims to assess the feasibility, efficacy, and safety of acupuncture for HT.

Methods: This is a randomized, black-controlled assessor-blinded pilot trial. A total of 60 patients will be recruited and divided into the experimental group (n = 30) or the control group (n = 30). The experimental group will undergo acupuncture therapy (penetration needling of Hand-Yangming meridian, PNHM) for 16 weeks, followed by a 16-week follow-up period, and the control group will first go through an observation period for 16 weeks, followed by a 16-week compensation PNHM therapy. The primary outcome will be the change of the concentrations of anti-thyroperoxidase antibodies (TPOAb), antithyroglobulin antibodies (TgAb), and thyroid hormone, including total thyroxine (FT), free thyroxine (FT), and thyroid-stimulating hormone (TSH). The secondary outcome measurements include the thyroid-related quality of life questionnaire short-form (ThyPRO-39), The Mos 36-item Short Form Health Survey (SF-36), and Hospital Anxiety and Depression Scale (HAD). Data collection will be performed before the start of the study (the baseline assessment) and at weeks 8, 16, 24, and 32.

Discussion: The study is designed to assess the feasibility and effectiveness of PNHM in reducing the thyroid antibody level and improving the quality of life of HT patients with hypothyroidism or subclinical hypothyroidism. Results of this trial will assist further analyses on whether the acupuncture treatment can alleviate symptoms for patients with HT.

Trial Registration: Acupuncture-Moxibustion Clinical Trial Registry AMCTR-IOR-19000308 ( ChiCTR1900026830 ). Registered on 23 October 2019.
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http://dx.doi.org/10.1186/s13063-021-05036-8DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7818748PMC
January 2021

The Underlying Mechanism of Pall. in Parkinson's Disease Based on a Network Pharmacology Approach.

Front Pharmacol 2020 23;11:581984. Epub 2020 Nov 23.

Department of Neurology, Xiyuan Hospital, China Academy of Chinese Medical Sciences, Beijing, China.

Parkinson's disease (PD) is the second most common neurodegenerative disease worldwide, yet as of currently, there is no disease-modifying therapy that could delay its progression. Pall. is the most frequently used herb in formulas for PD in Traditional Chinese Medicine and also a potential neuroprotective agent for neurodegenerative diseases, while its mechanisms remain poorly understood. In this study, we aim to explore the underlying mechanism of in treating PD utilizing a network pharmacology approach. The protein targets of ingredients and PD were first obtained from several databases. To clarify the key targets, a Protein-Protein-Interaction (PPI) network was constructed and analyzed on the String database, and then enrichment analysis was performed by the Metascape platform to determine the main Gene Ontology biological processes and Kyoto Encyclopedia of Genes and Genomes pathways. Finally, the Ingredient-Target-Pathway (I-T-P) network was constructed and analyzed by Cytoscape software. Six active ingredients of (kaempferol, ß-sitosterol, betulinic acid, palbinone, paeoniflorin and (+)-catechin) as well as six core targets strongly related to PD treatment [AKT1, interleukin-6, CAT, Tumor necrosis factor (TNF), CASP3, and PTGS2] were identified. The main pathways were shown to involve neuroactive ligand-receptor interaction, Calcium signaling pathway, PI3-Akt signaling pathway, TNF signaling pathway, and apoptosis signaling pathway. The main biological process included the regulation of neurotransmitter levels. may retard neurodegeneration by reducing neuroinflammation, inhibiting intrinsic and extrinsic apoptosis, and may improve motor and non-motor symptoms by regulating the levels of neurotransmitters. Our study has revealed the mechanism of in the treatment of PD and may contribute to novel drug development for PD.
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http://dx.doi.org/10.3389/fphar.2020.581984DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7768820PMC
November 2020

COPII mitigates ER stress by promoting formation of ER whorls.

Cell Res 2021 Feb 28;31(2):141-156. Epub 2020 Sep 28.

The State Key Laboratory of Membrane Biology, Tsinghua University-Peking University Joint Centre for Life Sciences, Beijing Frontier Research Center for Biological Structure, School of Life Sciences, Tsinghua University, Beijing, 100084, China.

Cells mitigate ER stress through the unfolded protein response (UPR). Here, we report formation of ER whorls as an effector mechanism of the ER stress response. We found that strong ER stress induces formation of ER whorls, which contain ER-resident proteins such as the Sec61 complex and PKR-like ER kinase (PERK). ER whorl formation is dependent on PERK kinase activity and is mediated by COPII machinery, which facilitates ER membrane budding to form tubular-vesicular ER whorl precursors. ER whorl precursors then go through Sec22b-mediated fusion to form ER whorls. We further show that ER whorls contribute to ER stress-induced translational inhibition by possibly modulating PERK activity and by sequestering translocons in a ribosome-free environment. We propose that formation of ER whorls reflects a new type of ER stress response that controls inhibition of protein translation.
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http://dx.doi.org/10.1038/s41422-020-00416-2DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8026990PMC
February 2021

Single-molecule in vitro reconstitution assay for kinesin-1-driven membrane dynamics.

Biophys Rev 2019 Jun 4;11(3):319-325. Epub 2019 May 4.

Institute for Biomedical Materials & Devices (IBMD), Faculty of Science, University of Technology Sydney, Ultimo, NSW, 2007, Australia.

Intracellular membrane dynamics, especially the nano-tube formation, plays important roles in vesicle transportation and organelle biogenesis. Regarding the regulation mechanisms, it is well known that during the nano-tube formation, motor proteins act as the driven force moving along the cytoskeleton, lipid composition and its associated proteins serve as the linkers and key mediators, and the vesicle sizes play as one of the important regulators. In this review, we summarized the in vitro reconstitution assay method, which has been applied to reconstitute the nano-tube dynamics during autophagic lysosomal regeneration (ALR) and the morphology dynamics during mitochondria network formation (MNF) in a mimic and pure in vitro system. Combined with the single-molecule microscopy, the advantage of the in vitro reconstitution system is to study the key questions at a single-molecule or single-vesicle level with precisely tuned parameters and conditions, such as the motor mutation, ion concentration, lipid component, ATP/GTP concentration, and even in vitro protein knockout, which cannot easily be achieved by in vivo or intracellular studies.
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http://dx.doi.org/10.1007/s12551-019-00531-4DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6557960PMC
June 2019

Kinesin 1 Drives Autolysosome Tubulation.

Dev Cell 2016 05;37(4):326-336

State Key Laboratory of Membrane Biology, Tsinghua University-Peking University Joint Center for Life Sciences, School of Life Sciences, Tsinghua University, Beijing 100084, China. Electronic address:

Autophagic lysosome reformation (ALR) plays an important role in maintaining lysosome homeostasis. During ALR, lysosomes are reformed by recycling lysosomal components from autolysosomes. The most noticeable step of ALR is autolysosome tubulation, but it is currently unknown how the process is regulated. Here, using an approach combining in vivo studies and in vitro reconstitution, we found that the kinesin motor protein KIF5B is required for autolysosome tubulation and that KIF5B drives autolysosome tubulation by pulling on the autolysosomal membrane. Furthermore, we show that KIF5B directly interacts with PtdIns(4,5)P2. Kinesin motors are recruited and clustered on autolysosomes via interaction with PtdIns(4,5)P2 in a clathrin-dependent manner. Finally, we demonstrate that clathrin promotes formation of PtdIns(4,5)P2-enriched microdomains, which are required for clustering of KIF5B. Our study reveals a mechanism by which autolysosome tubulation was generated.
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http://dx.doi.org/10.1016/j.devcel.2016.04.014DOI Listing
May 2016

Vesicle Size Regulates Nanotube Formation in the Cell.

Sci Rep 2016 04 7;6:24002. Epub 2016 Apr 7.

State Key Laboratory of Membrane Biology, Biodynamic Optical Imaging Center, School of Life Sciences, Peking University, Beijing 100871, China.

Intracellular membrane nanotube formation and its dynamics play important roles for cargo transportation and organelle biogenesis. Regarding the regulation mechanisms, while much attention has been paid on the lipid composition and its associated protein molecules, effects of the vesicle size has not been studied in the cell. Giant unilamellar vesicles (GUVs) are often used for in vitro membrane deformation studies, but they are much larger than most intracellular vesicles and the in vitro studies also lack physiological relevance. Here, we use lysosomes and autolysosomes, whose sizes range between 100 nm and 1 μm, as model systems to study the size effects on nanotube formation both in vivo and in vitro. Single molecule observations indicate that driven by kinesin motors, small vesicles (100-200 nm) are mainly transported along the tracks while a remarkable portion of large vesicles (500-1000 nm) form nanotubes. This size effect is further confirmed by in vitro reconstitution assays on liposomes and purified lysosomes and autolysosomes. We also apply Atomic Force Microscopy (AFM) to measure the initiation force for nanotube formation. These results suggest that the size-dependence may be one of the mechanisms for cells to regulate cellular processes involving membrane-deformation, such as the timing of tubulation-mediated vesicle recycling.
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http://dx.doi.org/10.1038/srep24002DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4823757PMC
April 2016

Dynamic tubulation of mitochondria drives mitochondrial network formation.

Cell Res 2015 Oct 24;25(10):1108-20. Epub 2015 Jul 24.

State Key Laboratory of Biomembrane and Membrane Biotechnology, Tsinghua University-Peking University Joint Center for Life Sciences, School of Life Sciences, Tsinghua University, Beijing 100084, China.

Mitochondria form networks. Formation of mitochondrial networks is important for maintaining mitochondrial DNA integrity and interchanging mitochondrial material, whereas disruption of the mitochondrial network affects mitochondrial functions. According to the current view, mitochondrial networks are formed by fusion of individual mitochondria. Here, we report a new mechanism for formation of mitochondrial networks through KIF5B-mediated dynamic tubulation of mitochondria. We found that KIF5B pulls thin, highly dynamic tubules out of mitochondria. Fusion of these dynamic tubules, which is mediated by mitofusins, gives rise to the mitochondrial network. We further demonstrated that dynamic tubulation and fusion is sufficient for mitochondrial network formation, by reconstituting mitochondrial networks in vitro using purified fusion-competent mitochondria, recombinant KIF5B, and polymerized microtubules. Interestingly, KIF5B only controls network formation in the peripheral zone of the cell, indicating that the mitochondrial network is divided into subzones, which may be constructed by different mechanisms. Our data not only uncover an essential mechanism for mitochondrial network formation, but also reveal that different parts of the mitochondrial network are formed by different mechanisms.
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http://dx.doi.org/10.1038/cr.2015.89DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4650629PMC
October 2015

Clathrin and phosphatidylinositol-4,5-bisphosphate regulate autophagic lysosome reformation.

Nat Cell Biol 2012 Sep 12;14(9):924-34. Epub 2012 Aug 12.

State Key Laboratory of Biomembrane and Membrane Biotechnology, Tsinghua University-Peking University Center for Life Sciences, School of Life Science, Tsinghua University, Beijing 100084, China.

Autophagy is a lysosome-based degradation pathway. During autophagy, lysosomes fuse with autophagosomes to form autolysosomes. Following starvation-induced autophagy, nascent lysosomes are formed from autolysosomal membranes through an evolutionarily conserved cellular process, autophagic lysosome reformation (ALR), which is critical for maintaining lysosome homeostasis. Here we report that clathrin and phosphatidylinositol-4,5-bisphosphate (PtdIns(4,5)P(2)) regulate ALR. Combining a screen of candidates identified through proteomic analysis of purified ALR tubules, and large-scale RNAi knockdown, we unveiled a tightly regulated molecular pathway that controls lysosome homeostasis, in which clathrin and PtdIns(4,5)P(2) are the central components. Our functional study demonstrates the central role of clathrin and its associated proteins in cargo sorting, phospholipid conversion, initiation of autolysosome tubulation, and proto-lysosome budding during ALR. Our data not only uncover a molecular pathway by which lysosome homeostasis is maintained through the ALR process, but also reveal unexpected functions of clathrin and PtdIns(4,5)P(2) in lysosome homeostasis.
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http://dx.doi.org/10.1038/ncb2557DOI Listing
September 2012

The promoting effect of byproducts from Irpex lacteus on subsequent enzymatic hydrolysis of bio-pretreated cornstalks.

Biotechnol Biofuels 2011 Oct 11;4(1):37. Epub 2011 Oct 11.

School of Life Science and Technology, Huazhong University of Science and Technology, Wuhan 430074, China.

Background: Irpex lacteus, a versatile lignin-degrading fungus with various extracellular enzymes, has been widely used for biological pretreatment. However, most studies have focused on the change of substrate structure after biological pretreatment, and the effect of these changes on the enzymatic hydrolysis, but the effect of byproducts from biological pretreatment process on subsequent enzymatic hydrolysis is not well understood.

Methods: We developed a biological pretreatment process with I. lacteus that can produce stimulatory byproducts that enhance the enzymatic hydrolysis of cornstalks.

Results: The maximum hydrolysis yield of glucan (82%) was obtained after pretreatment for 28 days. The maximum reducing sugar yield decreased from 313.5 to 200.1 mg/g raw cornstalks after water-soluble byproducts of biological pretreatment were removed from pretreated cornstalks. The effect of byproducts on enzymatic hydrolysis was also investigated. We found that the hydrolysis efficiency of commercial cellulase preparation on cornstalks could be improved by water extracts from bio-pretreated cornstalks with hydrolytic enzyme activity and iron-reducing activity.

Conclusion: The key finding suggested that byproducts from biological pretreatment play important roles in enhancing downstream hydrolysis, which might be attributable to hydrolytic enzymes and iron-reducing compounds produced by I. lacteus.
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http://dx.doi.org/10.1186/1754-6834-4-37DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3238224PMC
October 2011

Evaluation of white-rot fungi-assisted alkaline/oxidative pretreatment of corn straw undergoing enzymatic hydrolysis by cellulase.

J Biosci Bioeng 2010 Dec;110(6):660-4

College of Life Science and Technology, Huazhong University of Science and Technology, Wuluo Road 1037, Wuhan 430074, China.

In this study, the effects of biological treatment prior to alkaline/oxidative (A/O) pretreatment using three white-rot fungi (Ganoderma lucidum, Trametes versicolor and Echinodontium taxodii) were evaluated for the enzymatic hydrolysis of corn straw. Among these fungi, Echinodontium taxodii significantly enhanced the efficiency of chemical pretreatment. Subsequent to treatment of corn straw with Echinodontium taxodii for 15 days, the straw was subjected to digestion by 0.0016% NaOH and 3% H₂O₂ at room temperature for 24 h, which increased the reducing sugar yield by 50.7%. The hydrolysis model and kinetic parameters were determined from time course data collected throughout the hydrolysis. The initial hydrolysis rate, V₀, of the corn straw increased by 68.5% compared to A/O pretreatment alone, which resulted from an increase in the initial adsorption. The lignin content of the corn straw decreased more significantly after biological and A/O pretreatment than after A/O pretreatment alone. After 72 h of enzymatic hydrolysis, the adsorbed cellulase decreased by 24.8% (from 3.67 to 2.76 mg ml⁻¹) compared to A/O pretreatment alone. These results indicate that biological treatment improves the desorption of cellulase by enhancing delignification during A/O pretreatment.
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http://dx.doi.org/10.1016/j.jbiosc.2010.08.002DOI Listing
December 2010

Fungal treatment of cornstalks enhances the delignification and xylan loss during mild alkaline pretreatment and enzymatic digestibility of glucan.

Bioresour Technol 2010 Sep 18;101(17):6728-34. Epub 2010 Apr 18.

School of Life Science and Technology, Huazhong University of Science and Technology, Luoyu Road 1037, Wuhan 430074, China.

Fungal treatment with Irpex lacteus was used to enhance the delignification and xylan loss during mild alkaline pretreatment and subsequent enzymatic conversion in this research. The 15-day bio-treatment can modify the lignin structure and increase losses of lignin (from 75.67% to 80.00%) and xylan (from 40.68% to 51.37%) during alkaline pretreatment, making the enzymatic conversion more efficient. The high digestibility of glucan can be obtained after the bio-treatment and alkaline pretreatment at near room-temperature (30 degrees C), and the maximum digestibility increased 14% in comparison with that after the sole alkaline pretreatment. The bio-treatment enhanced delignification and glucan digestibility more significantly when the alkaline pretreatment was performed at lower severity. Additionally, Nuclei Growth model with a time-dependent rate constant can describe well the delignification and xylan loss. Results indicated that the bio-treatment increased the rate constant of initial reaction, but accelerated the decline of rate constant during alkaline pretreatment.
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http://dx.doi.org/10.1016/j.biortech.2010.03.119DOI Listing
September 2010