DOCSLIB.ORG

Inhibiting Heat Shock Protein 90 Protects Nucleus Pulposus-Derived Stem/Progenitor Cells from Compression-Induced Necroptosis and Apoptosis

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Inhibiting Heat Shock Protein 90 Protects Nucleus Pulposus-Derived Stem/Progenitor Cells from Compression-Induced Necroptosis and Apoptosis fcell-08-00685 August 7, 2020 Time: 16:35 # 1 ORIGINAL RESEARCH published: 07 August 2020 doi: 10.3389/fcell.2020.00685 Inhibiting Heat Shock Protein 90 Protects Nucleus Pulposus-Derived Stem/Progenitor Cells From Compression-Induced Necroptosis and Apoptosis Binwu Hu1†, Shuo Zhang1†, Weijian Liu1, Peng Wang1, Songfeng Chen2, Xiao Lv1, Deyao Shi1, Kaige Ma1, Baichuan Wang1, Yongchao Wu1 and Zengwu Shao1* Edited by: 1 Department of Orthopaedics, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Pavel Makarevich, Wuhan, China, 2 Department of Orthopaedic Surgery, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, China Lomonosov Moscow State University, Russia Reviewed by: Nucleus pulposus-derived stem/progenitor cells (NPSCs) provide novel prospects for Gelina Kopeina, the regeneration of degenerated intervertebral disc (IVD). Nevertheless, with aging Lomonosov Moscow State University, and degeneration of IVD, the frequency of NPSCs markedly decreases. Excessive Russia Xiaolei Zhang, cell death could be the main reason for declined frequency of NPSCs, however, Second Affiliated Hospital and Yuying the exact mechanisms remain elusive. Thus, the present study was undertaken to Children’s Hospital of Wenzhou Medical University, China explore the mechanisms of compression-induced NPSCs death, and the effects of heat *Correspondence: shock protein 90 (HSP90) on NPSCs survival. Here, we found that compression could Zengwu Shao trigger receptor-interacting protein kinase 1 (RIPK1)/receptor-interacting protein kinase [email protected] 3 (RIPK3)/mixed lineage kinase domain-like protein (MLKL)-mediated necroptosis of † These authors have contributed NPSCs. Furthermore, we found that elevated expression of HSP90 was involved in equally to this work compression-induced NPSCs death, and inhibiting HSP90 could dramatically attenuate Specialty section: compression-induced necroptosis of NPSCs via regulating the expression and activity This article was submitted to Molecular Medicine, of RIPK1/RIPK3/MLKL, and alleviating the mitochondrial dysfunction (mitochondrial a section of the journal membrane potential loss and ATP depletion) and oxidative stress [production of Frontiers in Cell and Developmental mitochondrial reactive oxygen species (ROS), cellular total ROS and malondialdehyde, Biology and downregulation of superoxide dismutase 2]. Besides necroptosis, compression- Received: 23 March 2020 Accepted: 06 July 2020 induced apoptosis of NPSCs was also attenuated by HSP90 inhibition. In addition, we Published: 07 August 2020 found that enhanced expression of HSP70 contributed to the cytoprotective effects of Citation: inhibiting HSP90. More encouragingly, our results demonstrated that inhibiting HSP90 Hu B, Zhang S, Liu W, Wang P, Chen S, Lv X, Shi D, Ma K, Wang B, could also mitigate the exhaustion of NPSCs in vivo. In conclusion, RIPK1/RIPK3/MLKL- Wu Y and Shao Z (2020) Inhibiting mediated necroptosis participates in compression-induced NPSCs death. Furthermore, Heat Shock Protein 90 Protects targeting HSP90 to simultaneously inhibit necroptosis and apoptosis of NPSCs might be Nucleus Pulposus-Derived Stem/Progenitor Cells From an efficient strategy for preventing the death of NPSCs, thus rescuing the endogenous Compression-Induced Necroptosis repair capacity of NP tissue. and Apoptosis. Front. Cell Dev. Biol. 8:685. Keywords: nucleus pulposus-derived stem/progenitor cells, heat shock protein 90, necroptosis, apoptosis, doi: 10.3389/fcell.2020.00685 intervertebral disc degeneration, compression Frontiers in Cell and Developmental Biology| www.frontiersin.org 1 August 2020| Volume 8| Article 685 fcell-08-00685 August 7, 2020 Time: 16:35 # 2 Hu et al. Inhibiting HSP90 Attenuates NPSCs Death INTRODUCTION survival through the activation of nuclear factor-kappa B, indirectly promote cell survival via HSP90-Akt complexes, and Intervertebral disc (IVD) degeneration (IVDD) has been widely modulate the intrinsic pathway of apoptosis (Arya et al., 2007). considered as the paramount etiology for low back pain, On the contrary, elevated expression of HSP90 could also exert leading to heavy socio-economic burden due to lost productivity pro-death roles under some circumstances (Hooven et al., 2004; and increasing health care costs (Maetzel and Li, 2002; Wang et al., 2011; Ma et al., 2015). Correspondingly, inhibiting Freemont, 2009). Accumulating evidence has demonstrated that HSP90 by specific inhibitors or siRNAs is able to downregulate dysregulation of the function and quantity of nucleus pulposus some cell death-related pathways, such as the IKK/IkB/p65 (NP) cells accounts for the initiation of IVDD (Buckwalter, 1995; pathway, as well as activate multiple cell survival pathways, Li et al., 2018b). Therefore, many treatments aiming at retarding including induction of heat shock protein 70 (HSP70) and the or reversing disc degeneration, including cell transplantation, phosphorylation of Akt, extracellular signal regulated kinase growth factors supplementation and gene therapy, mainly focus 1/2 and glycogen synthase kinase 3b, ultimately attenuating cell on reviving the function and quantity of NP cells (Smith et al., death in many pathological process (Wang et al., 2011; Alani 2011). However, these established methods have limitations more et al., 2014; Li J. et al., 2015; Liu Q. et al., 2016). Traditionally, or less (Zhao et al., 2007). Recently, many studies have proven the decreased cell death caused by HSP90 inhibition is mainly existence of NP-derived stem/progenitor cells (NPSCs) (Risbud attributed to reduced cell apoptosis, recent studies also reveal et al., 2007; Hu and He, 2018). It is reported that NPSCs could that RIPK1, RIPK3, and MLKL, the critical components of secrete extracellular matrix and differentiate toward NP cells necroptosis signaling pathway, are client proteins of HSP90 (Tao et al., 2015). Furthermore, the co-culture of NP cells and (Lewis et al., 2000; Li D. et al., 2015; Zhao et al., 2016). Inhibiting NPSCs could enhance cell proliferation and the expression of HSP90 could block necroptosis by modulating the stability and chondrogenic-associated genes under hypertonicity condition function of RIPK1, RIPK3, and MLKL (Yang and He, 2016). (Tao et al., 2013). Thus, NPSCs might provide novel prospects Thus, HSP90 could simultaneously regulate apoptosis and for the regeneration of degenerated IVD. However, with aging necroptosis, which might be a wonderful target for preventing and degeneration of IVD, the frequency of NPSCs markedly cell death. However, the roles of HSP90 in compression-induced decreases, indicating the exhaustion of NPSCs (Sakai et al., 2012). NPSCs death have not been determined yet. Excessive cell death could be the main reason for the exhaustion Mitochondrial dysfunction and reactive oxygen species (ROS) of NPSCs. Previous studies have demonstrated that many adverse are critical in the execution of necroptosis (Marshall and microenvironment factors, such as high osmolarity, low pH, Baines, 2014). Furthermore, they could also trigger cell apoptosis oxidative stress and mechanical loading, could induce NPSCs through mitochondrial apoptosis pathway (Feng et al., 2017). death (Hu and He, 2018; Nan et al., 2019). Nevertheless, the Our group has demonstrated that mitochondrial dysfunction precise mechanisms have not yet been fully elucidated. and oxidative stress contribute greatly to compression-induced Mechanical loading has long been recognized as the leading NP cells apoptosis and necroptosis (Ding et al., 2012; Chen cause of IVDD (Neidlinger-Wilke et al., 2014). Our previous et al., 2018). Furthermore, we found that compression could also studies demonstrate that programmed cell death, such as trigger mitochondrial dysfunction and oxidative stress of NPSCs, apoptosis and necroptosis, participates in compression-induced leading to cell apoptosis (Li et al., 2018a). Therefore, suppressing NP cells death (Ding et al., 2012; Chen et al., 2017). Meanwhile, mitochondrial dysfunction and oxidative stress might be efficient we found that compression could also trigger apoptosis of NPSCs methods for preventing compression-induced NPSCs apoptosis (Li et al., 2018a). However, it has not been fully elucidated and necroptosis. whether other types of programmed cell death are involved In present study, we first investigated the involvement in compression-induced NPSCs death. Necroptosis is a newly of necroptosis in compression-induced death of NPSCs. defined form of regulated necrosis, which is mainly mediated Subsequently, we determined the roles of HSP90 in compression- by receptor-interacting protein kinase 1 (RIPK1)/receptor- induced NPSCs death and explored relevant mechanisms. In interacting protein kinase 3 (RIPK3)/mixed lineage kinase addition, we also evaluated whether inhibiting HSP90 alleviated domain-like protein (MLKL) signaling pathway (Hu et al., 2018). the exhaustion of NPSCs in vivo. Our study provides novel It has been reported that necroptosis contributes to various insights into the mechanisms of compression-induced NPSCs pathophysiological conditions, including systemic inflammation, death as well as experimental evidence of a possible target for ischemic reperfusion injury and neurodegeneration (Zhou and protecting NPSCs. Yuan, 2014; Galluzzi et al., 2017). Therefore, unraveling the role of necroptosis in compression-induced NPSCs death, and its regulatory mechanisms might provide novel targets for MATERIALS AND METHODS preventing NPSCs death. Heat shock
Load more

Recommended publications
  • Targeting the LKB1 Tumor Suppressor
    Send Orders for Reprints to [email protected] 32 Current Drug Targets, 2014, 15, 32-52 Targeting the LKB1 Tumor Suppressor Rui-Xun Zhao and Zhi-Xiang Xu* Division of Hematology and Oncology, Comprehensive Cancer Center, University of Alabama at Birmingham, Birmingham, AL, USA Abstract: LKB1 (also known as serine-threonine kinase 11, STK11) is a tumor suppressor, which is mutated or deleted in Peutz-Jeghers syndrome (PJS) and in a variety of cancers. Physiologically, LKB1 possesses multiple cellular functions in the regulation of cell bioenergetics metabolism, cell cycle arrest, embryo development, cell polarity, and apoptosis. New studies demonstrated that LKB1 may also play a role in the maintenance of function and dynamics of hematopoietic stem cells. Over the past years, personalized therapy targeting specific genetic aberrations has attracted intense interests. Within this review, several agents with potential activity against aberrant LKB1 signaling have been discussed. Potential strate- gies and challenges in targeting LKB1 inactivation are also considered. Keywords: LKB1 (serine-threonine kinase 11, STK11), AMP-activated protein kinase (AMPK), tumor suppression, mutations, targeting therapeutics. INTRODUCTION THE BIOLOGICAL FUNCTIONS OF LKB1 The LKB1 gene, also known as serine-threonine kinase Cell Metabolism 11 (STK11), was first identified by Jun-ichi Nezu of Chugai About a decade ago, studies from three different groups Pharmaceuticals in 1996 in a screen aimed at identifying new established that LKB1 is the long-sought kinase that phos- kinases [1]. The human LKB1 gene has been mapped to phorylates AMPK [9-11]. AMPK is a heterotrimeric enzyme chromosome 19p13.3. The gene spans 23 kb and is com- complex consisting of a catalytic subunit and regulatory posed of nine coding exons and a noncoding exon [2].
    [Show full text]
  • A Computational Approach for Defining a Signature of Β-Cell Golgi Stress in Diabetes Mellitus
    Page 1 of 781 Diabetes A Computational Approach for Defining a Signature of β-Cell Golgi Stress in Diabetes Mellitus Robert N. Bone1,6,7, Olufunmilola Oyebamiji2, Sayali Talware2, Sharmila Selvaraj2, Preethi Krishnan3,6, Farooq Syed1,6,7, Huanmei Wu2, Carmella Evans-Molina 1,3,4,5,6,7,8* Departments of 1Pediatrics, 3Medicine, 4Anatomy, Cell Biology & Physiology, 5Biochemistry & Molecular Biology, the 6Center for Diabetes & Metabolic Diseases, and the 7Herman B. Wells Center for Pediatric Research, Indiana University School of Medicine, Indianapolis, IN 46202; 2Department of BioHealth Informatics, Indiana University-Purdue University Indianapolis, Indianapolis, IN, 46202; 8Roudebush VA Medical Center, Indianapolis, IN 46202. *Corresponding Author(s): Carmella Evans-Molina, MD, PhD ([email protected]) Indiana University School of Medicine, 635 Barnhill Drive, MS 2031A, Indianapolis, IN 46202, Telephone: (317) 274-4145, Fax (317) 274-4107 Running Title: Golgi Stress Response in Diabetes Word Count: 4358 Number of Figures: 6 Keywords: Golgi apparatus stress, Islets, β cell, Type 1 diabetes, Type 2 diabetes 1 Diabetes Publish Ahead of Print, published online August 20, 2020 Diabetes Page 2 of 781 ABSTRACT The Golgi apparatus (GA) is an important site of insulin processing and granule maturation, but whether GA organelle dysfunction and GA stress are present in the diabetic β-cell has not been tested. We utilized an informatics-based approach to develop a transcriptional signature of β-cell GA stress using existing RNA sequencing and microarray datasets generated using human islets from donors with diabetes and islets where type 1(T1D) and type 2 diabetes (T2D) had been modeled ex vivo. To narrow our results to GA-specific genes, we applied a filter set of 1,030 genes accepted as GA associated.
    [Show full text]
  • Supplementary Table 1. Genes Mapped in Core Cancer
    Supplementary Table 1. Genes mapped in core cancer pathways annotated by KEGG (Kyoto Encyclopedia of Genes and Genomes), MIPS (The Munich Information Center for Protein Sequences), BIOCARTA, PID (Pathway Interaction Database), and REACTOME databases. EP300,MAP2K1,APC,MAP3K7,ZFYVE9,TGFB2,TGFB1,CREBBP,MAP BIOCARTA TGFB PATHWAY K3,TAB1,SMAD3,SMAD4,TGFBR2,SKIL,TGFBR1,SMAD7,TGFB3,CD H1,SMAD2 TFDP1,NOG,TNF,GDF7,INHBB,INHBC,COMP,INHBA,THBS4,RHOA,C REBBP,ROCK1,ID1,ID2,RPS6KB1,RPS6KB2,CUL1,LOC728622,ID4,SM AD3,MAPK3,RBL2,SMAD4,RBL1,NODAL,SMAD1,MYC,SMAD2,MAP K1,SMURF2,SMURF1,EP300,BMP8A,GDF5,SKP1,CHRD,TGFB2,TGFB 1,IFNG,CDKN2B,PPP2CB,PPP2CA,PPP2R1A,ID3,SMAD5,RBX1,FST,PI KEGG TGF BETA SIGNALING PATHWAY TX2,PPP2R1B,TGFBR2,AMHR2,LTBP1,LEFTY1,AMH,TGFBR1,SMAD 9,LEFTY2,SMAD7,ROCK2,TGFB3,SMAD6,BMPR2,GDF6,BMPR1A,B MPR1B,ACVRL1,ACVR2B,ACVR2A,ACVR1,BMP4,E2F5,BMP2,ACVR 1C,E2F4,SP1,BMP7,BMP8B,ZFYVE9,BMP5,BMP6,ZFYVE16,THBS3,IN HBE,THBS2,DCN,THBS1, JUN,LRP5,LRP6,PPP3R2,SFRP2,SFRP1,PPP3CC,VANGL1,PPP3R1,FZD 1,FZD4,APC2,FZD6,FZD7,SENP2,FZD8,LEF1,CREBBP,FZD9,PRICKLE 1,CTBP2,ROCK1,CTBP1,WNT9B,WNT9A,CTNNBIP1,DAAM2,TBL1X R1,MMP7,CER1,MAP3K7,VANGL2,WNT2B,WNT11,WNT10B,DKK2,L OC728622,CHP2,AXIN1,AXIN2,DKK4,NFAT5,MYC,SOX17,CSNK2A1, CSNK2A2,NFATC4,CSNK1A1,NFATC3,CSNK1E,BTRC,PRKX,SKP1,FB XW11,RBX1,CSNK2B,SIAH1,TBL1Y,WNT5B,CCND1,CAMK2A,NLK, CAMK2B,CAMK2D,CAMK2G,PRKACA,APC,PRKACB,PRKACG,WNT 16,DAAM1,CHD8,FRAT1,CACYBP,CCND2,NFATC2,NFATC1,CCND3,P KEGG WNT SIGNALING PATHWAY LCB2,PLCB1,CSNK1A1L,PRKCB,PLCB3,PRKCA,PLCB4,WIF1,PRICK LE2,PORCN,RHOA,FRAT2,PRKCG,MAPK9,MAPK10,WNT3A,DVL3,R
    [Show full text]
  • The Activity and Stability of P56lck and TCR Signaling Do Not Depend on the Co-Chaperone Cdc37
    International Journal of Molecular Sciences Article The Activity and Stability of p56Lck and TCR Signaling Do Not Depend on the Co-Chaperone Cdc37 Sarah Kowallik 1,2, Andreas Kritikos 1,2, Matthias Kästle 1,2, Christoph Thurm 1,2, Burkhart Schraven 1,2 and Luca Simeoni 1,2,* 1 Institute of Molecular and Clinical Immunology, Otto-von-Guericke University, 39120 Magdeburg, Germany; [email protected] (S.K.); [email protected] (A.K.); [email protected] (M.K.); [email protected] (C.T.); [email protected] (B.S.) 2 Health Campus Immunology, Infectiology and Inflammation (GC-I3), Medical Faculty, Otto-von Guericke University, 39120 Magdeburg, Germany * Correspondence: [email protected]; Tel.: +49-391-67-17894 Abstract: Lymphocyte-specific protein tyrosine kinase (Lck) is a pivotal tyrosine kinase involved in T cell receptor (TCR) signaling. Because of its importance, the activity of Lck is regulated at different levels including phosphorylation of tyrosine residues, protein–protein interactions, and localization. It has been proposed that the co-chaperone Cdc37, which assists the chaperone heat shock protein 90 (Hsp90) in the folding of client proteins, is also involved in the regulation of the activity/stability of Lck. Nevertheless, the available experimental data do not clearly support this conclusion. Thus, we assessed whether or not Cdc37 regulates Lck. We performed experiments in which the expression of Cdc37 was either augmented or suppressed in Jurkat T cells. The results of our experiments indicated that neither the overexpression nor the suppression of Cdc37 affected Lck stability and activity.
    [Show full text]
  • Development and Validation of a Protein-Based Risk Score for Cardiovascular Outcomes Among Patients with Stable Coronary Heart Disease
    Supplementary Online Content Ganz P, Heidecker B, Hveem K, et al. Development and validation of a protein-based risk score for cardiovascular outcomes among patients with stable coronary heart disease. JAMA. doi: 10.1001/jama.2016.5951 eTable 1. List of 1130 Proteins Measured by Somalogic’s Modified Aptamer-Based Proteomic Assay eTable 2. Coefficients for Weibull Recalibration Model Applied to 9-Protein Model eFigure 1. Median Protein Levels in Derivation and Validation Cohort eTable 3. Coefficients for the Recalibration Model Applied to Refit Framingham eFigure 2. Calibration Plots for the Refit Framingham Model eTable 4. List of 200 Proteins Associated With the Risk of MI, Stroke, Heart Failure, and Death eFigure 3. Hazard Ratios of Lasso Selected Proteins for Primary End Point of MI, Stroke, Heart Failure, and Death eFigure 4. 9-Protein Prognostic Model Hazard Ratios Adjusted for Framingham Variables eFigure 5. 9-Protein Risk Scores by Event Type This supplementary material has been provided by the authors to give readers additional information about their work. Downloaded From: https://jamanetwork.com/ on 10/02/2021 Supplemental Material Table of Contents 1 Study Design and Data Processing ......................................................................................................... 3 2 Table of 1130 Proteins Measured .......................................................................................................... 4 3 Variable Selection and Statistical Modeling ........................................................................................
    [Show full text]
  • Hsp90 Interacts with the Bacterial Effector Nleh1
    pathogens Article Hsp90 Interacts with the Bacterial Effector NleH1 Miaomiao Wu and Philip R. Hardwidge * Department of Diagnostic Medicine/Pathobiology, Kansas State University, Manhattan, KS 66506, USA; [email protected] * Correspondence: [email protected]; Tel.: +1-785-532-2506 Received: 26 September 2018; Accepted: 11 November 2018; Published: 13 November 2018 Abstract: Enterohemorrhagic Escherichia coli (EHEC) utilizes a type III secretion system (T3SS) to inject effector proteins into host cells. The EHEC NleH1 effector inhibits the nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) pathway by reducing the nuclear translocation of the ribosomal protein S3 (RPS3). NleH1 prevents RPS3 phosphorylation by the IκB kinase-β (IKKβ). IKKβ is a central kinase in the NF-κB pathway, yet NleH1 only restricts the phosphorylation of a subset of the IKKβ substrates. We hypothesized that a protein cofactor might dictate this inhibitory specificity. We determined that heat shock protein 90 (Hsp90) interacts with both IKKβ and NleH1 and that inhibiting Hsp90 activity reduces RPS3 nuclear translocation. Keywords: E. coli; Hsp90; NleH1; RPS3 1. Introduction The nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) family of transcription factors regulates innate and adaptive immune responses. In addition to the well-characterized Rel family proteins [1], ribosomal protein S3 (RPS3) is a key non-Rel subunit and was identified as a “specifier” NF-κB component. RPS3 guides NF-κB to specific κB sites by increasing the affinity of the NF-κB p65 subunit for target gene promoters [2]. Activation of NF-κB signaling is initiated by external stimuli that activate the IκB kinase (IKK) complex.
    [Show full text]
  • Mammalian P50cdc37 Is a Protein Kinase-Targeting Subunit of Hsp90 That Binds and Stabilizes Cdk4
    Downloaded from genesdev.cshlp.org on October 5, 2021 - Published by Cold Spring Harbor Laboratory Press Mammalian p50Cdc37is a protein kinase- tar eting subunit of Hsp90 that binds ani stabilizes Cdk4 Lilia Stepanova,' Xiaohong ~eng,'Susan B. ~arker?and 7. Wade ~arper'?~ 'Verna and Marrs McLean Department of Biochemistry, 2~owardHughes Medical Institue, Baylor College of Medicine, Houston, Texas 77030 USA CDC37, an essential gene in Saccharomyces cerevisiae, interacts genetically with multiple protein kinases and is required for production of Cdc28plcyclin complexes through an unknown mechanism. We have identified mammalian p50~~~~~as a protein kinase-targeting subunit of the molecular chaperone Hsp90. Previously, p50 was observed in complexes with pp6OV-"" and Raf-1, but its identity and function have remained elusive. In mouse fibroblasts, a primary target of Cdc37 is Cdk4. This kinase is activated by D-type cyclins and functions in passage through GI. In insect cells, Cdc37 is sufficient to target Hsp90 to Cdk4 and both in vitro and in vivo, Cdc371Hsp90 associates preferentially with the fraction of Cdk4 not bound to D-type cyclins. Cdc37 is coexpressed with cyclin Dl in cells undergoing programmed proliferation in vivo, consistent with a positive role in cell cycle progression. Pharmacological inactivation of Cdc371Hsp90 function decreases the half-life of newly synthesized Cdk4, indicating a role for Cdc37IHsp90 in Cdk4 stabilization. This study suggests a general role for p5~Cdc37in signaling pathways dependent on intrinsically unstable protein kinases and reveals a previously unrecognized chaperone-dependent step in the production of Cdk4lcyclin D complexes. [Key Words: Cyclin-dependent kinase; D-type cyclin; molecular chaperone; Hsp90; p50Cdc37] Received March 22, 1996; revised version accepted April 30, 1996.
    [Show full text]
  • The Role of Heat Shock Proteins in Regulating Receptor Signal Transduction
    1521-0111/95/5/468–474$35.00 https://doi.org/10.1124/mol.118.114652 MOLECULAR PHARMACOLOGY Mol Pharmacol 95:468–474, May 2019 Copyright ª 2019 by The American Society for Pharmacology and Experimental Therapeutics MINIREVIEW—SPATIAL ORGANIZATION OF SIGNAL TRANSDUCTION The Role of Heat Shock Proteins in Regulating Receptor Signal Transduction John M. Streicher Department of Pharmacology, College of Medicine, University of Arizona, Tucson, Arizona Downloaded from Received September 20, 2018; accepted January 12, 2019 ABSTRACT Heat shock proteins (Hsp) are a class of stress-inducible with important impacts on endogenous and drug ligand re- proteins that mainly act as molecular protein chaperones. This sponses. Among these roles, Hsp90 in particular acts to molpharm.aspetjournals.org chaperone activity is diverse, including assisting in nascent maintain mature signaling kinases in a metastable conforma- protein folding and regulating client protein location and tion permissive for signaling activation. In this review, we will translocation within the cell. The main proteins within the Hsp focus on the roles of the Hsps, with a special focus on Hsp90, family, particularly Hsp70 and Hsp90, also have a highly diverse in regulating receptor signaling and subsequent physiologic and numerous set of protein clients, which when combined with responses. We will also explore potential means to manipulate the high expression levels of Hsp proteins (2%–6% of total Hsp function to improve receptor-targeted therapies. Overall, protein content) establishes these molecules as “central Hsps are important regulators of receptor signaling that are regulators” of cell protein physiology. Among the client receiving increasing interest and exploration, particularly as proteins, Hsps regulate numerous signal-transduction and Hsp90 inhibitors progress toward clinical approval for the receptor-regulatory kinases, and indeed directly regulate some treatment of cancer.
    [Show full text]
  • Restricting Direct Interaction of CDC37 with HSP90 Does Not Compromise Chaperoning of Client Proteins
    Oncogene (2015) 34, 15–26 & 2015 Macmillan Publishers Limited All rights reserved 0950-9232/15 www.nature.com/onc ORIGINAL ARTICLE Restricting direct interaction of CDC37 with HSP90 does not compromise chaperoning of client proteins JR Smith1, E de Billy1, S Hobbs1, M Powers1, C Prodromou2,3, L Pearl2,3, PA Clarke1 and P Workman1 The HSP90 molecular chaperone plays a key role in the maturation, stability and activation of its clients, including many oncogenic proteins. Kinases are a substantial and important subset of clients requiring the key cochaperone CDC37. We sought an improved understanding of protein kinase chaperoning by CDC37 in cancer cells. CDC37 overexpression in human colon cancer cells increased CDK4 protein levels, which was negated upon CDC37 knockdown. Overexpressing CDC37 increased CDK4 protein half- life and enhanced binding of HSP90 to CDK4, consistent with CDC37 promoting kinase loading onto chaperone complexes. Against expectation, expression of C-terminus-truncated CDC37 (DC-CDC37) that lacks HSP90 binding capacity did not affect kinase client expression or activity; moreover, as with wild-type CDC37 overexpression, it augmented CDK4-HSP90 complex formation. However, although truncation blocked binding to HSP90 in cells, DC-CDC37 also showed diminished client protein binding and was relatively unstable. CDC37 mutants with single and double point mutations at residues M164 and L205 showed greatly reduced binding to HSP90, but retained association with client kinases. Surprisingly, these mutants phenocopied wild-type CDC37 overexpression by increasing CDK4-HSP90 association and CDK4 protein levels in cells. Furthermore, expression of the mutants was sufficient to protect kinase clients CDK4, CDK6, CRAF and ERBB2 from depletion induced by silencing endogenous CDC37, indicating that CDC37’s client stabilising function cannot be inactivated by substantially reducing its direct interaction with HSP90.
    [Show full text]
  • The Human Gene Connectome As a Map of Short Cuts for Morbid Allele Discovery
    The human gene connectome as a map of short cuts for morbid allele discovery Yuval Itana,1, Shen-Ying Zhanga,b, Guillaume Vogta,b, Avinash Abhyankara, Melina Hermana, Patrick Nitschkec, Dror Friedd, Lluis Quintana-Murcie, Laurent Abela,b, and Jean-Laurent Casanovaa,b,f aSt. Giles Laboratory of Human Genetics of Infectious Diseases, Rockefeller Branch, The Rockefeller University, New York, NY 10065; bLaboratory of Human Genetics of Infectious Diseases, Necker Branch, Paris Descartes University, Institut National de la Santé et de la Recherche Médicale U980, Necker Medical School, 75015 Paris, France; cPlateforme Bioinformatique, Université Paris Descartes, 75116 Paris, France; dDepartment of Computer Science, Ben-Gurion University of the Negev, Beer-Sheva 84105, Israel; eUnit of Human Evolutionary Genetics, Centre National de la Recherche Scientifique, Unité de Recherche Associée 3012, Institut Pasteur, F-75015 Paris, France; and fPediatric Immunology-Hematology Unit, Necker Hospital for Sick Children, 75015 Paris, France Edited* by Bruce Beutler, University of Texas Southwestern Medical Center, Dallas, TX, and approved February 15, 2013 (received for review October 19, 2012) High-throughput genomic data reveal thousands of gene variants to detect a single mutated gene, with the other polymorphic genes per patient, and it is often difficult to determine which of these being of less interest. This goes some way to explaining why, variants underlies disease in a given individual. However, at the despite the abundance of NGS data, the discovery of disease- population level, there may be some degree of phenotypic homo- causing alleles from such data remains somewhat limited. geneity, with alterations of specific physiological pathways under- We developed the human gene connectome (HGC) to over- come this problem.
    [Show full text]
  • Three Novel Mutations of STK11 Gene in Chinese Patients with Peutz
    Tan et al. BMC Medical Genetics (2016) 17:77 DOI 10.1186/s12881-016-0339-6 CASE REPORT Open Access Three novel mutations of STK11 gene in Chinese patients with Peutz–Jeghers syndrome Hu Tan1, Libin Mei1, Yanru Huang1, Pu Yang1, Haoxian Li1, Ying Peng1, Chen Chen1,2, Xianda Wei1, Qian Pan1, Desheng Liang1* and Lingqian Wu1* Abstract Background: Peutz–Jeghers syndrome (PJS) is a rare autosomal dominant inherited disorder characterized by gastrointestinal (GI) hamartomatous polyps, mucocutaneous hyperpigmentation, and an increased risk of cancer. Mutations in the serine–threonine kinase 11 gene (SKT11) are the major cause of PJS. Case presentation: Blood samples were collected from six PJS families including eight patients. Mutation screening of STK11 gene was performed in these six families by Sanger sequencing and multiplex ligation- dependent probe amplification (MLPA) assay. Three novel mutations (c.721G > C, c.645_726del82, and del(exon2–5)) and three recurrent mutations (c.752G > A, c.545 T > C and del(exon1)) in STK11 were detected in six Chinese PJS families. Genotype-phenotype correlations suggested that truncating mutations trend to result in severe complications. Conclusion: These findings broaden the mutation spectrum of the STK11 gene and would help clinicians and genetic counselors provide better clinical surveillance for PJS patients, especially for ones carrying truncating mutation. Keywords: Peutz–Jeghers syndrome (PJS), Serine-threonine kinase 11 (STK11), Truncating mutation, Severe complication Background as a cause of PJS in 1998 [3, 4]. The gene, 23 kb in size, Peutz–Jeghers syndrome (PJS) is a rare autosomal dom- consists of nine coding exon and one non-coding exon inant inherited disorder characterized by gastrointestinal and encodes a 433-amino acid protein, which consists of (GI) hamartomatous polyps, mucocutaneous hyperpig- three domains: the N-terminal non-catalytic domain, the mentation of the lips, buccal mucosa, and digits.
    [Show full text]
  • Targeting the Hsp90-Cdc37-Client Protein Interaction to Disrupt Hsp90 Chaperone Machinery Ting Li1, Hu-Lin Jiang2, Yun-Guang Tong3,4 and Jin-Jian Lu1*
    Li et al. Journal of Hematology & Oncology (2018) 11:59 https://doi.org/10.1186/s13045-018-0602-8 REVIEW Open Access Targeting the Hsp90-Cdc37-client protein interaction to disrupt Hsp90 chaperone machinery Ting Li1, Hu-Lin Jiang2, Yun-Guang Tong3,4 and Jin-Jian Lu1* Abstract Heat shock protein 90 (Hsp90) is a critical molecular chaperone protein that regulates the folding, maturation, and stability of a wide variety of proteins. In recent years, the development of Hsp90-directed inhibitors has grown rapidly, and many of these inhibitors have entered clinical trials. In parallel, the functional dissection of the Hsp90 chaperone machinery has highlighted the activity disruption of Hsp90 co-chaperone as a potential target. With the roles of Hsp90 co-chaperones being elucidated, cell division cycle 37 (Cdc37), a ubiquitous co-chaperone of Hsp90 that directs the selective client proteins into the Hsp90 chaperone cycle, shows great promise. Moreover, the Hsp90-Cdc37-client interaction contributes to the regulation of cellular response and cellular growth and is more essential to tumor tissues than normal tissues. Herein, we discuss the current understanding of the clients of Hsp90-Cdc37, the interaction of Hsp90-Cdc37-client protein, and the therapeutic possibilities of targeting Hsp90-Cdc37-client protein interaction as a strategy to inhibit Hsp90 chaperone machinery to present new insights on alternative ways of inhibiting Hsp90 chaperone machinery. Keywords: Hsp90 chaperone machinery, Cdc37, Kinase client, Protein interaction Background chloroplast HSP90C, mitochondrial TNFR-associated protein, Heat shock protein 90 (Hsp90) is a critically conserved and bacterial high-temperature protein G [2, 8]. In this protein and one of the major molecular chaperones within review,weusethetermHsp90torefertotheseHsp90 eukaryotic cells [1].
    [Show full text]