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Article: Tat-haFGF

Meng, Tian / Cao, Qin / Lei, Peng / Bush, Ashley I / Xiang, Qi / Su, Zhijian / He, Xiang / Rogers, Jack T / Chiu, Ing-Ming / Zhang, Qihao / Huang, Yadong

Molecular therapy. Nucleic acids

2017 Volume 7, Page(s) 439–452

Abstract: ... in previous studies, yet its mechanism is still uncertain. Here we report that the efficacy of Tat-haFGF ...

Abstract	Acid fibroblast growth factor (aFGF) has shown neuroprotection in Alzheimer's disease (AD) models in previous studies, yet its mechanism is still uncertain. Here we report that the efficacy of Tat-haFGF
Language	English
Publishing date	2017-05-10
Publishing country	United States
Document type	Journal Article
ZDB-ID	2662631-7
ISSN	2162-2531
ISSN	2162-2531
DOI	10.1016/j.omtn.2017.05.004
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Article ; Online: HIV TAT-mediated microglial senescence

Annadurai Thangaraj / Ernest T. Chivero / Ashutosh Tripathi / Seema Singh / Fang Niu / Ming-Lei Guo / Prakash Pillai / Palsamy Periyasamy / Shilpa Buch

Redox Biology, Vol 40, Iss , Pp 101843- (2021)

Role of SIRT3-dependent mitochondrial oxidative stress

2021

Abstract: ... study was aimed at exploring the role of HIV TAT protein in mediating microglial mitochondrial ... that exposure of mouse primary microglial cells (mPMs) to HIV TAT protein resulted in a senescence-like ... of proinflammatory cytokines and decreased telomerase activity. Additionally, exposure of mPMs to HIV TAT also ...

Abstract	The advent of combined antiretroviral treatment (cART) as a treatment for HIV-1 infection has not only resulted in a dramatic decrease in the peripheral viral load but has also led to increased life expectancy of the infected individuals. Paradoxically, increased lifespan is accompanied with higher prevalence of age-related comorbidities, including HIV-associated neurocognitive disorders (HAND). Present study was aimed at exploring the role of HIV TAT protein in mediating microglial mitochondrial oxidative stress, ultimately resulting in neuroinflammation and microglial senescence. Our findings demonstrated that exposure of mouse primary microglial cells (mPMs) to HIV TAT protein resulted in a senescence-like phenotype, that was characterized by elevated expression of both p16 and p21 proteins, increased numbers of senescence-associated-β-galactosidase positive cells, augmented cell-cycle arrest, increased release of proinflammatory cytokines and decreased telomerase activity. Additionally, exposure of mPMs to HIV TAT also resulted downregulation of SIRT3 with a concomitant increase in mitochondrial oxidative stress. Dual luciferase reporter assay identified miR-505 as a novel target of SIRT3, which was upregulated in mPMs exposed to HIV TAT. Furthermore, transient transfection of mPMs with either the SIRT3 plasmid or miRNA-505 inhibitor upregulated the expression of SIRT3 and mitochondrial antioxidant enzymes, with a concomitant decrease in microglial senescence. These in vitro findings were also validated in the prefrontal cortices and striatum of HIV transgenic rats as well as cART-treated HIV-infected individuals. In summary, this study underscores a yet undiscovered novel mechanism(s) underlying HIV TAT-mediated induction of senescence phenotype in microglia, involving the miR-505-SIRT3 axis-mediated induction of mitochondrial oxidative stress.
Keywords	Mitochondria stress ; SIRT3 ; HIV TAT ; Microglial activation ; Neuroinflammation ; miRNA ; Medicine (General) ; R5-920 ; Biology (General) ; QH301-705.5
Language	English
Publishing date	2021-04-01T00:00:00Z
Publisher	Elsevier
Document type	Article ; Online
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Article ; Online: Role of Dysregulated Autophagy in HIV Tat, Cocaine, and cART Mediated NLRP3 Activation in Microglia.

Singh, Seema / Thangaraj, Annadurai / Chivero, Ernest T / Guo, Ming-Lei / Periyasamy, Palsamy / Buch, Shilpa

Journal of neuroimmune pharmacology : the official journal of the Society on NeuroImmune Pharmacology

2023 Volume 18, Issue 3, Page(s) 327–347

Abstract: ... persistence low levels of HIV proteins such as Transactivator of transcription (Tat ... The combined effects of HIV Tat, drugs of abuse, and cART can thus create a toxic milieu in the CNS. The present study ... investigated the combinatorial effects of HIV-Tat, cocaine, and cART on autophagy and NLRP3 inflammasome ...

Abstract	Despite the ability of combination antiretroviral therapy (cART) to suppress viremia, there is persistence low levels of HIV proteins such as Transactivator of transcription (Tat) in the central nervous system (CNS), contributing to glial activation and neuroinflammation. Accumulating evidence also implicates the role of drugs of abuse in exacerbating neurological complications associated with HIV-1. The combined effects of HIV Tat, drugs of abuse, and cART can thus create a toxic milieu in the CNS. The present study investigated the combinatorial effects of HIV-Tat, cocaine, and cART on autophagy and NLRP3 inflammasome activation. We selected a combination of three commonly used cART regimens: tenofovir, emtricitabine, and dolutegravir. Our results demonstrated that exposure of mouse primary microglia (MPMs) to these agents-HIV Tat (25 ng/ml), cocaine (1 μM), and cART (1 μM each) resulted in upregulation of autophagy markers: Beclin1, LC3B-II, and SQSTM1 with impaired lysosomal functioning involving increased lysosomal pH, decreased LAMP2 and cathepsin D, ultimately leading to dysregulated autophagy. Our findings also demonstrated activation of the NLRP3 signaling in microglia exposed to these agents. We further demonstrated that gene silencing of key autophagy protein BECN1 significantly blocked NLRP3-mediated activation of microglia. Silencing of NLRP3, however, failed to block HIV Tat, cocaine, and cART-mediated dysregulation of the autophagy-lysosomal axis; these in vitro phenomena were also validated in vivo using iTat mice administered cocaine and cART. This study thus underscores the cooperative effects of HIV Tat, cocaine, and cART in exacerbating microglial activation involving dysregulated autophagy and activation of the NLRP3 inflammasome signaling.
MeSH term(s)	Mice ; Animals ; Microglia ; NLR Family, Pyrin Domain-Containing 3 Protein/metabolism ; Cocaine/pharmacology ; Inflammasomes/metabolism ; Trans-Activators/metabolism ; Trans-Activators/pharmacology ; Autophagy ; tat Gene Products, Human Immunodeficiency Virus/metabolism ; HIV Infections/metabolism
Chemical Substances	NLR Family, Pyrin Domain-Containing 3 Protein ; Cocaine (I5Y540LHVR) ; Inflammasomes ; Trans-Activators ; tat Gene Products, Human Immunodeficiency Virus
Language	English
Publishing date	2023-05-06
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't ; Research Support, N.I.H., Extramural
ZDB-ID	2227405-4
ISSN	1557-1904 ; 1557-1890
ISSN (online)	1557-1904
ISSN	1557-1890
DOI	10.1007/s11481-023-10063-0
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Article ; Online: Development and Validation of a Liquid Chromatography-Tandem Mass Spectrometry Method for Sensitive Analysis of Residual Protein Tat Bh1-101 in Lentiviral Vectors for Gene Therapy.

Lu, Yan / Zhang, Chao-Xuan / Rodrigues, Patrick / Yang, Lei / Shafer, Aaron / Rankovic, Zoran / Fazio, Frank

Human gene therapy

2023 Volume 34, Issue 21-22, Page(s) 1162–1171

Abstract: ... Some LV vectors are produced using stable 293T packaging cell lines, which includes gag-pol-rev-tat and ... virus-glycoprotein. Transactivating factor (transactivator of transcription [Tat]) is a regulatory ... protein that drastically increases the efficiency of lentiviral transcription. Residual analysis of Tat is ...

Abstract	Lentiviral (LV) vector-based gene therapy is gaining popularity for treating a wide range of diseases. Various LV vectors are being developed for transducing cells in cellular gene therapy at St. Jude. Some LV vectors are produced using stable 293T packaging cell lines, which includes gag-pol-rev-tat and virus-glycoprotein. Transactivating factor (transactivator of transcription [Tat]) is a regulatory protein that drastically increases the efficiency of lentiviral transcription. Residual analysis of Tat is critical for gene vector quality and safety. In this work, we developed a highly sensitive liquid chromatography-tandem mass spectrometry method for analysis of residual Tat in Lentivirus as an alternative to enzyme-linked immunosorbent assay. Residual Tat in LV can be accurately quantified with high specificity with a limit of detection of 0.3 ng/mL.
MeSH term(s)	Transduction, Genetic ; Trans-Activators/genetics ; Tandem Mass Spectrometry ; Lentivirus/genetics ; Genetic Vectors/genetics ; Genetic Therapy
Chemical Substances	Trans-Activators
Language	English
Publishing date	2023-10-26
Publishing country	United States
Document type	Journal Article
ZDB-ID	1028152-6
ISSN	1557-7422 ; 1043-0342
ISSN (online)	1557-7422
ISSN	1043-0342
DOI	10.1089/hum.2023.030
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Zs.A 2988: Show issues

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Article ; Online: HIV-1 subtype B Tat enhances NOTCH3 signaling in astrocytes to mediate oxidative stress, inflammatory response, and neuronal apoptosis.

Gao, Lin / Sun, Weixi / Zhang, Dongmei / Shang, Yanxing / Li, Li / Tao, Wenhua / Zhang, Lei / Liu, Hongbin

Journal of neurovirology

2023 Volume 29, Issue 4, Page(s) 479–491

Abstract: ... Transactivator of transcription (Tat) induces oxidative stress and inflammatory response in astrocytes, thereby ... upregulated during subtype B or C Tat expression in HEB astroglial cells. Moreover, bioinformatics analysis ... encephalitis patients was higher than that of HIV control patients. Of note, subtype B Tat, rather than subtype ...

Abstract	NOTCH receptors are relevant to multiple neurodegenerative diseases. However, the roles and mechanisms of NOTCH receptors in HIV-associated neurocognitive disorder (HAND) remain largely unclear. Transactivator of transcription (Tat) induces oxidative stress and inflammatory response in astrocytes, thereby leading to neuronal apoptosis in the central nervous system. We determined that NOTCH3 expression was upregulated during subtype B or C Tat expression in HEB astroglial cells. Moreover, bioinformatics analysis of the Gene Expression Omnibus (GEO) dataset revealed that NOTCH3 mRNA expression in the frontal cortex tissues of HIV encephalitis patients was higher than that of HIV control patients. Of note, subtype B Tat, rather than subtype C Tat, interacted with the extracellular domain of the NOTCH3 receptor, thus activating NOTCH3 signaling. Downregulation of NOTCH3 attenuated subtype B Tat-induced oxidative stress and reactive oxygen species generation. In addition, we demonstrated that NOTCH3 signaling facilitated subtype B Tat-activated NF-κB signaling pathway, thereby mediating pro-inflammatory cytokines IL-6 and TNF-α production. Furthermore, downregulation of NOTCH3 in HEB astroglial cells protected SH-SY5Y neuronal cells from astrocyte-mediated subtype B Tat neurotoxicity. Taken together, our study clarifies the potential role of NOTCH3 in subtype B Tat-induced oxidative stress and inflammatory response in astrocytes, which could be a novel therapeutic target for the relief of HAND.
MeSH term(s)	Humans ; Astrocytes/metabolism ; HIV-1/genetics ; HIV-1/metabolism ; Trans-Activators/metabolism ; Receptor, Notch3/genetics ; Receptor, Notch3/metabolism ; tat Gene Products, Human Immunodeficiency Virus/genetics ; tat Gene Products, Human Immunodeficiency Virus/metabolism ; Neuroblastoma/metabolism ; Signal Transduction ; NF-kappa B/genetics ; NF-kappa B/metabolism ; HIV Infections/genetics ; HIV Infections/metabolism ; Oxidative Stress ; Apoptosis ; Basic Helix-Loop-Helix Transcription Factors/metabolism
Chemical Substances	Trans-Activators ; Receptor, Notch3 ; tat Gene Products, Human Immunodeficiency Virus ; NF-kappa B ; Basic Helix-Loop-Helix Transcription Factors ; NOTCH3 protein, human
Language	English
Publishing date	2023-06-26
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	1283265-0
ISSN	1538-2443 ; 1355-0284
ISSN (online)	1538-2443
ISSN	1355-0284
DOI	10.1007/s13365-023-01151-1
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Zs.A 4426: Show issues

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Article ; Online: Corrigendum to “HIV TAT-mediated microglial senescence

Annadurai Thangaraj / Ernest T. Chivero / Ashutosh Tripathi / Seema Singh / Fang Niu / Ming-Lei Guo / Prakash Pillai / Palsamy Periyasamy / Shilpa Buch

Redox Biology, Vol 44, Iss , Pp 101909- (2021)

Role of SIRT3-dependent mitochondrial oxidative stress” [Redox Biol. 40 (2021) 101843]

2021

Keywords	Medicine (General) ; R5-920 ; Biology (General) ; QH301-705.5
Language	English
Publishing date	2021-08-01T00:00:00Z
Publisher	Elsevier
Document type	Article ; Online
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Article ; Online: Loading of "cocktail siRNAs" into extracellular vesicles via TAT-DRBD peptide for the treatment of castration-resistant prostate cancer.

Diao, Yanjun / Wang, Gangqiang / Zhu, Bingbing / Li, Zhuo / Wang, Shan / Yu, Lijuan / Li, Rui / Fan, Weixiao / Zhang, Yue / Zhou, Lei / Yang, Liu / Hao, Xiaoke / Liu, Jiayun

Cancer biology & therapy

2022 Volume 23, Issue 1, Page(s) 163–172

Abstract: ... we report a novel strategy using polycationic membrane-penetrating peptide TAT to encapsulate siRNAs ... into EVs. Three TAT peptides were co-expressed with DRBD as 3TD fusion protein. The sequence-independent ... and the significantly downregulated expression of three genes suggested the potential of TAT ...

Abstract	Extracellular vesicles (EVs) are cell-derived, membranous nanoparticles that mediate intercellular communication by transferring biomolecules between cells. As natural vehicles, EVs may exhibit higher delivery efficiency, lower immunogenicity, and better compatibility than existing RNA carriers. A major limitation of their therapeutic use is the shortage of efficient, robust, and scalable methods to load siRNA of interest. Here, we report a novel strategy using polycationic membrane-penetrating peptide TAT to encapsulate siRNAs into EVs. Three TAT peptides were co-expressed with DRBD as 3TD fusion protein. The sequence-independent binding of DRBD facilitates multiplex genes targeting of mixed siRNAs. Functional assays for siRNA-mediated gene silencing of CRPC were performed after engineered EVs treatment. EVs were isolated using differential centrifugation from WPMY-1 cell culture medium. The increase of merged yellow fluorescence in the engineered EVs showed by TIRFM and the decrease in zeta potential absolute values certified the co-localization of siRNA with EVs, which indicated that siRNA had been successfully delivered into WPMY-1 EVs. qRT-PCR analysis revealed that the mRNA level of FLOH1, NKX3, and DHRS7 was dramatically decreased when cells were treated with engineered EVs loaded with siRNAs mixtures relative to the level of untreated cells. Western and flow cytometry results indicate that delivery of siRNA mixtures by engineered EVs can effectively downregulate AR expression and induce LNCaP-AI cell apoptosis. The uptake efficiency of the EVs and the significantly downregulated expression of three genes suggested the potential of TAT as efficient siRNA carriers by keeping the function of the cargoes.
MeSH term(s)	Extracellular Vesicles/metabolism ; Humans ; Male ; Nanoparticles ; Oxidoreductases/metabolism ; Peptides/metabolism ; Prostatic Neoplasms, Castration-Resistant/genetics ; Prostatic Neoplasms, Castration-Resistant/metabolism ; Prostatic Neoplasms, Castration-Resistant/therapy ; RNA, Small Interfering/genetics ; RNA, Small Interfering/metabolism
Chemical Substances	Peptides ; RNA, Small Interfering ; DHRS7 protein, human (EC 1.-) ; Oxidoreductases (EC 1.-)
Language	English
Publishing date	2022-02-15
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2146305-0
ISSN	1555-8576 ; 1538-4047
ISSN (online)	1555-8576
ISSN	1538-4047
DOI	10.1080/15384047.2021.2024040
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Zs.A 5887: Show issues

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Article: Passage of Magnetic Tat-Conjugated Fe

Zhao, Xueqin / Shang, Ting / Zhang, Xiaodan / Ye, Ting / Wang, Dajin / Rei, Lei

Nanoscale research letters

2016 Volume 11, Issue 1, Page(s) 451

Abstract: Delivery of diagnostic or therapeutic agents across the blood-brain barrier (BBB) remains a major challenge of brain disease treatment. Magnetic nanoparticles are actively being developed as drug carriers due to magnetic targeting and subsequently ... ...

Abstract	Delivery of diagnostic or therapeutic agents across the blood-brain barrier (BBB) remains a major challenge of brain disease treatment. Magnetic nanoparticles are actively being developed as drug carriers due to magnetic targeting and subsequently reduced off-target effects. In this paper, we developed a magnetic SiO
Language	English
Publishing date	2016-10-10
Publishing country	United States
Document type	Journal Article
ZDB-ID	2253244-4
ISSN	1556-276X ; 1931-7573
ISSN (online)	1556-276X
ISSN	1931-7573
DOI	10.1186/s11671-016-1676-2
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Article ; Online: Functional expression and purification of recombinant full-length human ATG7 protein with HIV-1 Tat peptide in Escherichia coli.

Dong, Guofu / Zhao, Xi / Guo, Junwang / Ma, Lei / Zhou, Hongmei / Liu, Qi / Zhao, Xuelong / Wang, Changzhen / Wu, Ke

Protein expression and purification

2021 Volume 182, Page(s) 105844

Abstract: ... challenging. Previous studies have demonstrated that the HIV-1 virus-encoded Tat peptide ('GRKKRRQRRR' ... expressed using the pET28b-Tat expression vector in the Escherichia coli BL21 (DE3) strain. Recombinant ... plasmids. In contrast, the solubility of Tat-tagged hATG7 increased significantly with prolonged time ...

Abstract	The human autophagy-related protein ATG7 (hATG7), an E1-like ubiquitin enzyme, activates two ubiquitin-like proteins, LC3 (Atg8) and Atg12, and promotes autophagosome formation. While hATG7 plays an essential role for the autophagy conjugation system, the production of full-length functional hATG7 in bacterial systems remains challenging. Previous studies have demonstrated that the HIV-1 virus-encoded Tat peptide ('GRKKRRQRRR') can increase the yield and solubility of heterologous proteins. Here, functional full-length hATG7 was expressed using the pET28b-Tat expression vector in the Escherichia coli BL21 (DE3) strain. Recombinant hATG7 protein aggregated as inclusion bodies while expressed with widely used prokaryotic expression plasmids. In contrast, the solubility of Tat-tagged hATG7 increased significantly with prolonged time compared to Tat-free hATG7. The recombinant proteins were purified to >90% homogeneity under native conditions with a single step of affinity chromatography purification. The results of in vitro pull-down and LC3B-I lipidation assays showed that Tat-tagged hATG7 directly interacted with LC3B-I and promoted LC3B-I lipidation, suggesting that Tat-tagged hATG7 has significant catalytic activity. Overall, this study provides a novel method for improving the functional expression of full-length hATG7 in bacterial systems by fusion with the Tat peptide, a process which may be applied in future studies of hATG7 structure and function.
MeSH term(s)	Autophagy-Related Protein 7/biosynthesis ; Autophagy-Related Protein 7/chemistry ; Autophagy-Related Protein 7/genetics ; Autophagy-Related Protein 7/isolation & purification ; Escherichia coli/genetics ; Escherichia coli/metabolism ; Gene Expression ; HIV-1/genetics ; Humans ; Recombinant Fusion Proteins/biosynthesis ; Recombinant Fusion Proteins/chemistry ; Recombinant Fusion Proteins/genetics ; Recombinant Fusion Proteins/isolation & purification ; tat Gene Products, Human Immunodeficiency Virus/biosynthesis ; tat Gene Products, Human Immunodeficiency Virus/chemistry ; tat Gene Products, Human Immunodeficiency Virus/genetics ; tat Gene Products, Human Immunodeficiency Virus/isolation & purification
Chemical Substances	Recombinant Fusion Proteins ; tat Gene Products, Human Immunodeficiency Virus ; ATG7 protein, human (EC 6.2.1.45) ; Autophagy-Related Protein 7 (EC 6.2.1.45)
Language	English
Publishing date	2021-02-13
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	1055455-5
ISSN	1096-0279 ; 1046-5928
ISSN (online)	1096-0279
ISSN	1046-5928
DOI	10.1016/j.pep.2021.105844
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Zs.A 3084: Show issues

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Article ; Online: Antioxidant Fusion Protein SOD1-Tat Increases the Engraftment Efficiency of Total Bone Marrow Cells in Irradiated Mice.

Bei, Ting / Cao, Xusong / Liu, Yun / Li, Jinmei / Luo, Haihua / Huang, Lin / Tian, Tian / Li, Lei / Jiang, Yong

Molecules (Basel, Switzerland)

2021 Volume 26, Issue 11

Abstract: ... superoxide dismutase 1 (SOD1) fused with the cell-penetrating peptide (CPP), the trans-activator of transcription (Tat ... cells and bone marrow cells in vitro. Furthermore, irradiated mice transplanted with SOD1-Tat-treated ...

Abstract	Total body irradiation is a standard procedure of bone marrow transplantation (BMT) which causes a rapid increase in reactive oxygen species (ROS) in the bone marrow microenvironment during BMT. The increase in ROS reduces the engraftment ability of donor cells, thereby affecting the bone marrow recovery of recipients after BMT. In the early weeks following transplantation, recipients are at high risk of severe infection due to weakened hematopoiesis. Thus, it is imperative to improve engraftment capacity and accelerate bone marrow recovery in BMT recipients. In this study, we constructed recombinant copper/zinc superoxide dismutase 1 (SOD1) fused with the cell-penetrating peptide (CPP), the trans-activator of transcription (Tat), and showed that this fusion protein has penetrating ability and antioxidant activity in both RAW264.7 cells and bone marrow cells in vitro. Furthermore, irradiated mice transplanted with SOD1-Tat-treated total bone marrow donor cells showed an increase in total bone marrow engraftment capacity two weeks after transplantation. This study explored an innovative method for enhancing engraftment efficiency and highlights the potential of CPP-SOD1 in ROS manipulation during BMT.
MeSH term(s)	Animals ; Antioxidants/metabolism ; Antioxidants/pharmacology ; Bone Marrow Cells/cytology ; Bone Marrow Cells/drug effects ; Bone Marrow Transplantation ; Cell-Penetrating Peptides/genetics ; Cell-Penetrating Peptides/metabolism ; Cells, Cultured ; Gene Products, tat/genetics ; Gene Products, tat/metabolism ; Male ; Mice ; Mice, Inbred C57BL ; Reactive Oxygen Species ; Recombinant Fusion Proteins/metabolism ; Recombinant Fusion Proteins/pharmacology ; Superoxide Dismutase-1/genetics ; Superoxide Dismutase-1/metabolism ; Whole-Body Irradiation
Chemical Substances	Antioxidants ; Cell-Penetrating Peptides ; Gene Products, tat ; Reactive Oxygen Species ; Recombinant Fusion Proteins ; Superoxide Dismutase-1 (EC 1.15.1.1)
Language	English
Publishing date	2021-06-03
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	1413402-0
ISSN	1420-3049 ; 1431-5165 ; 1420-3049
ISSN (online)	1420-3049
ISSN	1431-5165 ; 1420-3049
DOI	10.3390/molecules26113395
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