DOCSLIB.ORG

The Hsp90 Inhibitor Geldanamycin Selectively Sensitizes Bcr-Abl

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
The Hsp90 Inhibitor Geldanamycin Selectively Sensitizes Bcr-Abl Leukemia (2001) 15, 1537–1543 2001 Nature Publishing Group All rights reserved 0887-6924/01 $15.00 www.nature.com/leu The Hsp90 inhibitor geldanamycin selectively sensitizes Bcr-Abl-expressing leukemia cells to cytotoxic chemotherapy MV Blagosklonny1, T Fojo1, KN Bhalla3, J-S Kim1, JB Trepel1, WD Figg1, Y Rivera2 and LM Neckers2 Departments of 1Developmental Therapeutics and 2Cell and Cancer Biology, Medicine Branch, National Cancer Institute, NIH, Bethesda and Rockville, MD, USA; and 3Moffitt Cancer Center, Tampa, FL, USA The Bcr-Abl fusion protein drives leukemogenesis and can ren- Materials and methods der leukemia cells resistant to conventional chemotherapy. Geldanamycin (GA), a drug which destabilizes Hsp90- associated proteins, depletes cells of Bcr-Abl, an Hsp90 client, Cell lines and reagents but not of Abl. Both HL60 cells transfected with Bcr-Abl and naturally Ph1-positive K562 leukemia cells are resistant to most HL60 and Jurkat, human leukemia cell lines, were obtained cytotoxic drugs, but were found to be sensitive to GA. Further- from American Type Culture Collection (Manassas, VA, USA). more, GA sensitized Bcr-Abl-expressing cells to doxorubicin (DOX) and paclitaxel (PTX). In contrast, in parental HL60 cells, HL60-Bcr-Abl, a Bcr-Abl stable transfected HL-60 cells, were 8,16 90 nM GA inhibited PARP cleavage, nuclear fragmentation, and described previously. Paclitaxel (Taxol), was Bristol-Myers cell death caused by 500 ng/ml DOX. Like GA, STI 571 (an (Princeton, NJ, USA) product. Adriamycin (doxorubicin) was inhibitor of the Abl kinase) sensitized Bcr-Abl-expressing cells obtained from Sigma (St Louis, MO, USA) and dissolved in to DOX. Unlike GA, STI 571 did not antagonize the cytotoxic DMSO as a 2 mg/ml stock solution. STI 571 was kindly pro- effects of DOX in parental HL60 cells. These results indicate vided by Novartis Pharma (East Hanover, NJ, USA). The pro- that sensitization of Bcr-Abl-expressing cells, but not desensi- tization of HL60 cells, depends on inhibition of Bcr-Abl. Thus, teasome inhibitor PS-341 was a kind gift from Dr Peter Elliott, GA differentially affects leukemia cells depending on their Bcr- Proscript, Inc. (Cambridge, MA, USA). Abl expression and selectively increases apoptosis in Bcr-Abl- expressing cells. Leukemia (2001) 15, 1537–1543. Keywords: oncogenes; Bcr-Abl; chemotherapy; cytoprotection; gendanamycin; HSP90 Immunoblot analysis Cells were lysed and soluble proteins were harvested in TNES buffer (50 mm Tris HCl pH 7.5, 100 mm NaCl, 2 mm EDTA, Introduction 1mm sodium orthovanadate, 1% (v/v) NP40) containing pro- tease inhibitors (20 ␮g/ml aprotinin, 20 ␮g/ml leupeptin, 1 mm Chronic myelogenous leukemia (CML) is characterized by a PMSF). Proteins were resolved with SDS-PAGE as previously reciprocal t(9;22) chromosomal translocation, known as the described.15,17 Immunoblotting was performed using rabbit Philadelphia chromosome, that fuses the truncated Bcr gene polyclonal anti-human PARP (Upstate Biotechnology, Lake to the truncated c-Abl.1,2 Bcr-Abl is also often found in adult Placid, NY, USA), for Bcr using monoclonal anti-human Bcr acute lymphocytic leukemia (ALL).3 The Bcr-Abl gene enco- antibodies N 2 (Oncogene Science, Calbiochem, San Diego, des for character chimeric p210 and p185 proteins.4 These CA, USA). are constitutively active tyrosine kinases which may induce resistance to apoptotic cell death.5–12 A number of protein kinases, including Raf-1, Src, and ErbB, depend on association with the chaperone protein, heat-shock MTT assay protein 90 (Hsp90), for proper function and stability. The benzoquinone ansamycin geldanamycin (GA) binds to Hsp90 Fifteen thousand cells were plated in 0.1 ml in 96-well flat and specifically inhibits this chaperone’s function, resulting in bottom plates and then exposed to tested agents (final volume ␮ degradation of HSP90-associated proteins.13,14 The chimeric 0.2 ml per well). At the indicated time, 20 l of 5 mg/ml MTT BCR-ABL exists in a complex with Hsp90, and GA causes solution in PBS was added to each well for 4 h. After removal ␮ degradation of BCR-ABL after 3–5 h of treatment.15 Since Bcr- of the medium, 170 l of DMSO was added to each well to Abl expression is associated with resistance to cytotoxic dissolve the formazan crystals. The absorbance at 540 nm was agents, we hypothesized that treatment of Bcr-Abl-expressing determined using a Biokinetics plate reader (Bio-Tek Instru- cells with GA would restore their sensitivity to chemothera- ments, Inc, Winooski, VT, USA). Triplicate wells were assayed peutic agents. Indeed our results confirm this prediction. for each condition and standard deviations were determined. Importantly, while sensitizing Bcr-Abl-expressing cells, GA protected parental cells from cytotoxic effects of doxo- rubicin (DOX). Number of dead and live cells Cells were plated in 24-well plates in 1 ml of medium, or in 96-well plates in 0.2 ml, and were treated with drugs. After the indicated time, cells were counted in triplicate on a Coul- ter Z1 cell counter (Hialeah, FL, USA). In addition, cells were Correspondence: MV Blagosklonny, Medicine Branch, Bldg 10, R incubated with trypan blue and the numbers of blue (dead) 12N226, NIH, Bethesda, MD 20892, USA cells and transparent (live) cells were counted in a hemocyto- Received 30 January 2001; accepted 26 June 2001 meter. Sensitization of Bcr-Abl-expressing cells MV Blagosklonny et al 1538 Figure 2 Reversal of resistance/sensitivity ratios in HL60 and K562 cells by GA. Effects of increasing concentrations of GA on the cytotox- icity caused by 500 nm DOX in either K562 (closed triangles) or HL60 (open triangles) cells. MTT assay was performed after 3 days as described in Materials and methods. At GA = 0 nM axis X, the values represent the cytotoxicity of 500 ng/ml alone. Results were calculated as the percent of values obtained with untreated cells and represent mean ± s.d. Figure 1 Bcr-Abl downregulation and cytotoxic effects of GA in K562 cells. (a) K562 cells were incubated with indicated concen- trations of GA and then immunoblot for Bcr-Abl was performed as Results described in Materials and methods. Multiple lower molecular bands may represent products of BCR-ABL degradation in GA-treated cells. (b) K562 cells and HL60 cells (shown for comparison) were incubated Pharmacological depletion of Bcr-Abl and cytotoxicity with GA. MTT assay was performed after 3 days, as described in Materials and methods. Results were calculated as the percent of In agreement with previous report,15 treatment of K562 cells values obtained with untreated cells and represent mean ± s.d. with 30–810 nm GA completely depleted Bcr-Abl protein (Figure 1a). Similarly, 30 nm of GA caused a near-maximal growth inhibition of K562 cells (Figure 1b). Previous studies have shown that K562 leukemia cells expressing Bcr-Abl are Cell cycle analysis resistant to numerous therapeutic agents including anti-CD95 mAb, high-dose AraC, etoposide, paclitaxel, actinomycin D, 5,8,9,16,18,19 Cells were washed with PBS and resuspended in 75% ethanol staurosporine, and proteasome inhibitors. In con- in PBS and kept at 4°C for at least 30 min. Prior to analysis, trast, GA was more cytotoxic in K562 cells than in HL60 cells cells were washed again with PBS and resuspended and incu- (Figure 1b). bated for 30 min in propidium iodide staining solution con- taining 0.05 mg/ml propidium iodide (Sigma), 1 mm EDTA, 0.1% Triton-X-100 and 1 mg/ml RNAse A in PBS. The suspen- GA sensitized K562 cells but protected HL60 cells sion was then passed through a nylon mesh filter and ana- lyzed on a Becton Dickinson FACScan (Franklin Lakes, NJ, K562 cells were much more resistant to doxorubicin (DOX) USA). than were HL60 cells. (60% and 4% survival, respectively; see Figure 2, X-axis: GA = 0). Addition of 10–100 nm GA decreased survival of DOX-treated K562 cells. If sensitization by GA was due to depletion of Bcr-Abl, this effect should not Nuclear fragmentation assay be observed in HL60 cells. In fact, GA did not sensitize HL60 cells (Figure 2). Unexpectedly, GA rendered these cells more Cells were incubated with drugs for the indicated time. Cells resistant to DOX. Sensitization of K562 coupled with desensit- were washed with PBS, pelleted on to glass slides in a cyto- ization of HL60 resulted in the reversal of their relative sensi- centrifuge, fixed with 90% ethanol with 10% glacial acid and tivities to DOX in the presence of GA (Figure 2). stained with DAPI as described previously.18 Nuclei were When added alone as well as with either PTX or DOX, GA visualized under UV microscopy. depleted p210BCR-ABL in K562 cells (Figure 3a). Also, PTX and Leukemia Sensitization of Bcr-Abl-expressing cells MV Blagosklonny et al 1539 Figure 3 Combinations of drugs with GA. K562 cells (a) and HL60 cells (b) were treated with 90 nm GA, 500 ng/ml DOX, 100 nm PTX or their combinations, as indicated. After 16 h, cells were lysed and immunoblots for PARP and BCR were performed as described in Materials and methods. K562 cells (c) and HL60 cells (d) were treated as described in (a) and (b). MTT assay was performed after 48 h as described in Materials and methods. Results were calculated as the percent of values obtained with untreated cells and represent mean ± s.d. DOX slightly downregulated Bcr-Abl. This downregulation of gated combinations of GA with either PS-341, an inhibitor of Bcr-Abl occurred before apoptosis, since PARP was still intact the proteasome, or DOX (Figure 6). As expected, HL60-Bcr- 16 h after the treatment (Figure 3a). In HL60 cells, Bcr levels Abl were more resistant to PS-341 than were the parental cells were not affected by GA (Figure 3b).
Load more

Recommended publications
  • C-Terminal HSP90 Inhibitors Block the HSP90:HIF-1Α Interaction and Inhibit the Cellular Hypoxic Response
    bioRxiv preprint doi: https://doi.org/10.1101/521989; this version posted January 24, 2019. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. C-terminal HSP90 Inhibitors Block the HSP90:HIF-1α Interaction and Inhibit the Cellular Hypoxic Response Nalin Katariaa, Bernadette Kerra, Samantha S. Zaiterb, Shelli McAlpineb and Kristina M Cooka† a. University of Sydney, Faculty of Medicine and Health, Charles Perkins Centre, Sydney, Australia. b. School of Chemistry, University of New South Wales, Sydney, Australia. † To whom correspondence should be addressed: Kristina M Cook, Charles Perkins Centre, University of Sydney, Sydney, NSW, Australia 2006; [email protected]; Tel: +61 286274858. Hypoxia Inducible Factor (HIF) is a transcription factor cancer cells known as a heat shock response (HSR)12. The activated by low oxygen, which is common in solid compounds also have poor selectivity for HSP9013,14. tumours. HIF controls the expression of genes involved in C-terminus inhibitors of HSP90 (SM molecules)11 act in a angiogenesis, chemotherapy resistance and metastasis. The selective manner, and in contrast to the N-terminus chaperone HSP90 (Heat Shock Protein 90) stabilizes the inhibitors, they do not induce a heat shock response13,15. subunit HIF-1α and prevents degradation. Previously Although the SM molecules block all co-chaperones that identified HSP90 inhibitors bind to the N-terminal pocket bind to the C-terminus of HSP90 and inhibit HSP90 of HSP90 which blocks binding to HIF-1α, and produces function16,17, there is no data discussing whether C- HIF-1α degradation.
    [Show full text]
  • Hsp90 Inhibitor As a Sensitizer of Cancer Cells to Different Therapies (Review)
    INTERNATIONAL JOURNAL OF ONCOLOGY 46: 907-926, 2015 Hsp90 inhibitor as a sensitizer of cancer cells to different therapies (Review) ZUZANA SOLÁROVÁ1, Ján MOJžiš1 and PETER SOLÁR2 1Department of Pharmacology, Faculty of Medicine, 2Laboratory of Cell Biology, Institute of Biology and Ecology, Faculty of Science, P.J. šafárik University, 040 01 Košice, Slovak Republic Received September 10, 2014; Accepted October 22, 2014 DOI: 10.3892/ijo.2014.2791 Abstract. Hsp90 is a molecular chaperone that maintains 1. Introduction the structural and functional integrity of various client proteins involved in signaling and many other functions of The cell responds to environmental stress by increasing cancer cells. The natural inhibitors, ansamycins influence the synthesis of several molecular chaperons known as heat shock Hsp90 chaperone function by preventing its binding to client proteins (Hsp). These proteins are cathegorized according to proteins and resulting in their proteasomal degradation. N- their molecular weight into five classes: small Hsp, Hsp60, and C-terminal inhibitors of Hsp90 and their analogues are Hsp70, Hsp90 and Hsp100. One of the most abundant widely tested as potential anticancer agents in vitro, in vivo molecular chaperones is Hsp90, which is a highly conserved as well as in clinical trials. It seems that Hsp90 competi- protein, whose association is required for the stability and tive inhibitors target different tumor types at nanomolar function of multiple-mutated, chimeric- and overexpressed- concentrations and might have therapeutic benefit. On the signaling proteins that promote the growth and/or survival contrary, some Hsp90 inhibitors increased toxicity and resis- of cancer cells (1). Hsp90 client proteins lack specific Hsp90 tance of cancer cells induced by heat shock response, and binding motifs and vary in terms of intracellular localization, through the interaction of survival signals, that occured as structure, and function.
    [Show full text]
  • Combination of Anti-Cancer Drugs with Molecular Chaperone Inhibitors
    International Journal of Molecular Sciences Review Combination of Anti-Cancer Drugs with Molecular Chaperone Inhibitors Maxim Shevtsov 1,2,3,4,5,6,* , Gabriele Multhoff 1 , Elena Mikhaylova 2, Atsushi Shibata 7 , Irina Guzhova 2 and Boris Margulis 2,* 1 Center for Translational Cancer Research, TUM (TranslaTUM), Technische Universität München (TUM), Klinikum rechts der Isar, Radiation Immuno Oncology, Einsteinstr. 25, 81675 Munich, Germany; gabriele.multhoff@tum.de 2 Institute of Cytology of the Russian Academy of Sciences (RAS), Tikhoretsky Ave., 4, St. Petersburg 194064, Russia; [email protected] (E.M.); [email protected] (I.G.) 3 Department of Biotechnology Pavlov First Saint Petersburg State Medical University, L’va Tolstogo str., 6/8, St. Petersburg 197022, Russia 4 Almazov National Medical Research Centre, Polenov Russian Scientific Research Institute of Neurosurgery, Mayakovskogo str., 12, St. Petersburg 191014, Russia 5 Department of Biomedical Cell Technologies Far Eastern Federal University, Russky Island, Vladivostok 690000, Russia 6 National Center for Neurosurgery, Turan Ave., 34/1, Nur-Sultan 010000, Kazakhstan 7 Signal Transduction Program, Gunma University Initiative for Advanced Research (GIAR), Gunma University, Maebashi, Gunma 371-8511, Japan; [email protected] * Correspondence: [email protected] (M.S.); [email protected] (B.M.) Received: 31 August 2019; Accepted: 22 October 2019; Published: 24 October 2019 Abstract: Most molecular chaperones belonging to heat shock protein (HSP) families are known to protect cancer cells from pathologic, environmental and pharmacological stress factors and thereby can hamper anti-cancer therapies. In this review, we present data on inhibitors of the heat shock response (particularly mediated by the chaperones HSP90, HSP70, and HSP27) either as a single treatment or in combination with currently available anti-cancer therapeutic approaches.
    [Show full text]
  • The Hsp90 Chaperone Network Modulates Candida Virulence Traits
    HHS Public Access Author manuscript Author ManuscriptAuthor Manuscript Author Trends Microbiol Manuscript Author . Author Manuscript Author manuscript; available in PMC 2018 October 01. Published in final edited form as: Trends Microbiol. 2017 October ; 25(10): 809–819. doi:10.1016/j.tim.2017.05.003. The Hsp90 Chaperone Network Modulates Candida Virulence Traits Teresa R. O'Meara, Nicole Robbins, and Leah E. Cowen* Department of Molecular Genetics, University of Toronto, Toronto, Ontario M5G 1M1, Canada Abstract Hsp90 is a conserved molecular chaperone that facilitates the folding and function of client proteins. Hsp90 function is dynamically regulated by interactions with co-chaperones and by post- translational modifications. In the fungal pathogen Candida albicans, Hsp90 enables drug resistance and virulence by stabilizing diverse signal transducers. Here, we review studies that have unveiled regulators of Hsp90 function, as well as downstream effectors that govern the key virulence traits of morphogenesis and drug resistance. We highlight recent work mapping the Hsp90 genetic network in C. albicans under diverse environmental conditions, and how these interactions provide insight into circuitry important for drug resistance, morphogenesis, and virulence. Ultimately, elucidating the Hsp90 chaperone network will aid in the development of therapeutics to treat fungal disease. Keywords Candida albicans; stress response; Hsp90; virulence; development; drug resistance Hsp90 enables temperature-dependent morphogenesis, drug resistance and virulence in fungal pathogens For fungal pathogens, the ability to grow at human physiological temperatures is a requirement for successful colonization and infection [1-4]. Of the estimated 1.5 million fungal species, only approximately 300 can cause disease in humans and only a handful are common human pathogens [5].
    [Show full text]
  • Hsp90 Inhibitors for the Treatment of Chronic Myeloid Leukemia
    Hindawi Publishing Corporation Leukemia Research and Treatment Volume 2015, Article ID 757694, 16 pages http://dx.doi.org/10.1155/2015/757694 Review Article Hsp90 Inhibitors for the Treatment of Chronic Myeloid Leukemia Kalubai Vari Khajapeer and Rajasekaran Baskaran Department of Biochemistry and Molecular Biology, School of Life Sciences, Pondicherry University, Pondicherry 605014, India Correspondence should be addressed to Rajasekaran Baskaran; [email protected] Received 26 August 2015; Revised 11 November 2015; Accepted 12 November 2015 Academic Editor: Massimo Breccia Copyright © 2015 K. V. Khajapeer and R. Baskaran. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Chronic myeloid leukemia (CML) is a hematological malignancy that arises due to reciprocal translocation of 3 sequences from c-Abelson (ABL) protooncogene of chromosome 9 with 5 sequence of truncated break point cluster region (BCR) on chromosome 22. BCR-ABL is a functional oncoprotein p210 that exhibits constitutively activated tyrosine kinase causing genomic alteration of hematopoietic stem cells. BCR-ABL specific tyrosine kinase inhibitors (TKIs) successfully block CML progression. However, drug resistance owing to BCR-ABL mutations and overexpression is still an issue. Heat-shock proteins (Hsps) function as molecular chaperones facilitating proper folding of nascent polypeptides. Their increased expression under stressful conditions protects cells by stabilizing unfolded or misfolded peptides. Hsp90 is the major mammalian protein and is required by BCR-ABL for stabilization and maturation. Hsp90 inhibitors destabilize the binding of BCR-ABL protein thus leading to the formation of heteroprotein complex that is eventually degraded by the ubiquitin-proteasome pathway.
    [Show full text]
  • Geldanamycin and Its 17-Allylamino-17-Demethoxy
    [CANCER RESEARCH 63, 3241–3246, June 15, 2003] Geldanamycin and its 17-Allylamino-17-Demethoxy Analogue Antagonize the Action of Cisplatin in Human Colon Adenocarcinoma Cells: Differential Caspase Activation as a Basis for Interaction1 Irina A. Vasilevskaya,2 Tatiana V. Rakitina, and Peter J. O’Dwyer University of Pennsylvania Cancer Center, Philadelphia, Pennsylvania 19104 ABSTRACT mal degradation (10, 11). The ability of GA to deplete Raf1, ErbB2, and mutant p53 in breast cancer cells was found to correlate with its Several chaperone-binding drugs based on geldanamycin (GA) have antiproliferative activity (12), making it a plausible candidate for use been synthesized, and one of them, 17-allylamino-17-demethoxygeldana- mycin (17-AAG), is being developed in the clinic. Interest in the use of in cancer treatment. However, GA caused liver toxicity in preclinical 17-AAG in combination with cytotoxic drugs led us to study both GA and studies (13). A search for more tolerable derivatives has yielded 17-AAG with cisplatin (DDP) in the human colon adenocarcinoma cell 17-AAG, a less toxic analogue that retains the tumoricidal features of lines HT29 and HCT116. We performed isobologram analysis of combi- GA. Like its parent compound, 17-AAG inhibits several signaling nations of DDP with GA or 17-AAG in these cell lines using the standard pathways through binding to Hsp 90, which results in destabilization 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay to of signaling complexes and degradation of its client proteins by the evaluate cell survival. In HCT116, the effects of GA and 17-AAG with proteasome in a variety of cell lines (14–16).
    [Show full text]
  • A Novel Hsp90 Inhibitor Activates Compensatory Heat Shock Protein Responses and Autophagy and Alleviates Mutant A53T A-Synuclein Toxicity S
    Supplemental material to this article can be found at: http://molpharm.aspetjournals.org/content/suppl/2015/09/24/mol.115.101451.DC1 1521-0111/88/6/1045–1054$25.00 http://dx.doi.org/10.1124/mol.115.101451 MOLECULAR PHARMACOLOGY Mol Pharmacol 88:1045–1054, December 2015 Copyright ª 2015 by The American Society for Pharmacology and Experimental Therapeutics A Novel Hsp90 Inhibitor Activates Compensatory Heat Shock Protein Responses and Autophagy and Alleviates Mutant A53T a-Synuclein Toxicity s Rui Xiong, Wenbo Zhou, David Siegel, Russell R. A. Kitson, Curt R. Freed, Christopher J. Moody, and David Ross Department of Pharmaceutical Sciences, Skaggs School of Pharmacy and Pharmaceutical Sciences (R.X., D.S., D.R.), and Department of Medicine, Division of Clinical Pharmacology and Toxicology (W.Z., C.R.F.), University of Colorado Anschutz Medical Campus, Aurora, Colorado; and School of Chemistry, University of Nottingham, Nottingham, United Kingdom (R.R.A.K., Downloaded from C.J.M.) Received August 20, 2015; accepted September 16, 2015 ABSTRACT A potential cause of neurodegenerative diseases, including 2,5-diphenyl-tetrazolium bromide (MTT), and apoptosis assays. molpharm.aspetjournals.org Parkinson’s disease (PD), is protein misfolding and aggregation Meanwhile, 19-phenyl-GA retained the ability to induce autophagy that in turn leads to neurotoxicity. Targeting Hsp90 is an and potentially protective heat shock proteins (HSPs) such as attractive strategy to halt neurodegenerative diseases, and Hsp70 and Hsp27. We found that transduction of A53T, but not benzoquinone ansamycin (BQA) Hsp90 inhibitors such as gelda- wild type (WT) a-synuclein, induced toxicity in SH-SY5Y cells.
    [Show full text]
  • Cell Surface GRP94 As a Novel Emerging Therapeutic Target for Monoclonal Antibody Cancer Therapy
    cells Review Cell Surface GRP94 as a Novel Emerging Therapeutic Target for Monoclonal Antibody Cancer Therapy Ji Woong Kim † , Yea Bin Cho † and Sukmook Lee * Biopharmaceutical Chemistry Major, School of Applied Chemistry, Kookmin University, Seoul 02707, Korea; [email protected] (J.W.K.); [email protected] (Y.B.C.) * Correspondence: [email protected]; Tel.: +82-2-910-6763 † These authors contributed equally to this work. Abstract: Glucose-regulated protein 94 (GRP94) is an endoplasmic reticulum (ER)-resident member of the heat shock protein 90 (HSP90) family. In physiological conditions, it plays a vital role in regulating biological functions, including chaperoning cellular proteins in the ER lumen, maintaining calcium homeostasis, and modulating immune system function.‘Recently, several reports have shown the functional role and clinical relevance of GRP94 overexpression in the progression and metastasis of several cancers. Therefore, the current review highlights GRP94’s physiological and pathophysiological roles in normal and cancer cells. Additionally, the unmet medical needs of small chemical inhibitors and the current development status of monoclonal antibodies specifically targeting GRP94 will be discussed to emphasize the importance of cell surface GRP94 as an emerging therapeutic target in monoclonal antibody therapy for cancer. Keywords: GRP94; cancer; therapeutic target; therapy; monoclonal antibody Citation: Kim, J.W.; Cho, Y.B.; Lee, S. Cell Surface GRP94 as a Novel 1. Introduction Emerging Therapeutic Target for Cancer, the second leading cause of death behind cardiovascular disease, remains a Monoclonal Antibody Cancer major global public health problem [1]. Overall, the burden of cancer incidence and mortal- Therapy. Cells 2021, 10, 670.
    [Show full text]
  • HSP90 Inhibitor 17-AAG Selectively Eradicates Lymphoma Stem Cells
    Published OnlineFirst June 29, 2012; DOI: 10.1158/0008-5472.CAN-11-3600 Cancer Tumor and Stem Cell Biology Research HSP90 Inhibitor 17-AAG Selectively Eradicates Lymphoma Stem Cells Bryan Newman2, Yan Liu1, Hsiu-Fang Lee2, Duxin Sun2, and Yin Wang1 Abstract Cancer stem cells (CSC; also called tumor-initiating cells) comprise tumor cell subpopulations that preserve the properties of quiescence, self-renewal, and differentiation of normal stem cells. In addition, CSCs are therapeutically important because of their key contributions toward drug resistance. The hypoxia- inducible transcription factor HIF1a is critical for CSC maintenance in mouse lymphoma. In this study, we showed that low concentrations of the HSP90 inhibitor 17-AAG eliminate lymphoma CSCs in vitro and in vivo by disrupting the transcriptional function of HIF1a, a client protein of HSP90. 17-AAG preferentially induced apoptosis and eliminated the colony formation capacity of mouse lymphoma CSCs and human acute myeloid leukemia (AML) CSCs. However, low concentrations of 17-AAG failed to eliminate highly proliferative lymphoma and AML cells (non-CSCs), in which the AKT-GSK3 signaling pathway is constitutively active. The heat shock transcription factor HSF1 is highly expressed in non-CSCs, but it was weakly expressed in lymphoma CSCs. However, siRNA-mediated attenuation of HSF1 abrogated the colony formation ability of both lymphoma and AML CSCs. This study supports the use of 17-AAG as a CSC targeting agent and, in addition, shows that HSF1 is an important target for elimination of both CSCs and non-CSCs in cancer. Cancer Res; 72(17); 1–11. Ó2012 AACR. Introduction HSP90 inhibitors is a derivative of the geldanamycin antibiotic, Molecular chaperone proteins function to ensure the proper 17-N-allylamino-17-demethoxy geldanamycin(17-AAG;ref.12).
    [Show full text]
  • Resveratrol Induces Apoptosis in K562 Cells
    View metadata, citation and similar papers at core.ac.uk brought to you by CORE provided by Publications of the IAS Fellows ResveratrolBlackwell Publishing Ltd induces apoptosis in K562 (chronic myelogenous leukemia) cells by targeting a key survival protein, heat shock protein 70 Prabir K. Chakraborty,1 Soumyajit Banerjee Mustafi,1 Sudipto Ganguly,2 Mitali Chatterjee2 and Sanghamitra Raha1,3 1Crystallography and Molecular Biology Division, Saha Institute of Nuclear Physics, 1/AF Bidhan Nagar, Kolkata-700064; 2Department of Pharmacology, Institute of Post Graduate Medical Education and Research, 244B Acharya JC Bose Road, Kolkata-700020, India (Received October 15, 2007/Revised February 10, 2008/Accepted February 12, 2008/Online publication April 21, 2008) Chronic myelogenous leukemia (CML) is a myeloproliferative disease reports has documented a lack of patient response to imatinib. associated with a characteristic chromosomal translocation called This might be due to point mutations resulting in impaired the Philadelphia chromosome. This results in the expression of the imatinib binding at the kinase activity site of Bcr-abl.(11–13) Bcr-Abl fusion protein, a constitutively active protein tyrosine Therefore, the search for other novel targets and new strategies kinase. Although there are a few treatment options with Bcr-Abl for the management of CML continues. Numerous natural kinase inhibitors, drug resistance is often encountered. One of the products are under investigation for their clinical efficacy in major obstacles in overcoming drug resistance in CML is the high prevention and treatment of a wide array of diseases including endogenous levels of heat shock protein 70 (Hsp70). Resveratrol cancer.(14,15) Among these, trans-resveratrol (trans-3,4′,5-tri is a phytoalexin produced by several plants.
    [Show full text]
  • HSP90 Inhibitor AUY922 Induces Cell Death by Disruption of the Bcr-Abl, Jak2 and HSP90 Signaling Network Complex in Leukemia Cells
    www.impactjournals.com/Genes&Cancer Genes & Cancer, Vol. 6 (1-2), January 2015 HSP90 inhibitor AUY922 induces cell death by disruption of the Bcr-Abl, Jak2 and HSP90 signaling network complex in leukemia cells Wenjing Tao1,*, Sandip N. Chakraborty1,*, Xiaohong Leng1, Helen Ma1, and Ralph B. Arlinghaus1 1 Department of Translational Molecular Pathology, University of Texas M.D. Anderson Cancer Center, Houston, TX, USA. * These author contributed equally to this work Correspondence to: Ralph B. Arlinghaus, email: [email protected] Keywords: HSP90, Bcr-Abl, CML, apoptosis, gel filtration Received: December 29, 2014 Accepted: January 28, 2015 Published: January 29, 2015 This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. ABSTRACT The Bcr-Abl protein is an important client protein of heat shock protein 90 (HSP90). We evaluated the inhibitory effects of the HSP90 ATPase inhibitor AUY922 on 32D mouse hematopoietic cells expressing wild-type Bcr-Abl (b3a2, 32Dp210) and mutant Bcr-Abl imatinib (IM)-resistant cell lines. Western blotting results of fractions from gel filtration column chromatography of 32Dp210 cells showed that HSP90 together with Bcr-Abl, Jak2 Stat3 and several other proteins co-eluted in peak column fractions of a high molecular weight network complex (HMWNC). Co-IP results showed that HSP90 directly bound to Bcr-Abl, Jak2, Stat 3 and Akt. The associations between HSP90 and Bcr-Abl or Bcr-Abl kinase domain mutants (T315I and E255K) were interrupted by AUY922 treatment. Tyrosine phosphorylation of Bcr-Abl showed a dose-dependent decrease in 32Dp210T315I following AUY922 treatment for 16h.
    [Show full text]
  • Cytotoxicity Activity of Geldanamycin Derivatives Against Various Cancer Cell Lines
    Journal of Applied Pharmaceutical Science Vol. 10(06), pp 012-021, June, 2020 Available online at http://www.japsonline.com DOI: 10.7324/JAPS.2020.10603 ISSN 2231-3354 Cytotoxicity activity of geldanamycin derivatives against various cancer cell lines Thongchai Taechowisan1*, Tipparat Samsawat1, Winyou Puckdee1, Waya S. Phutdhawong2 1Department of Microbiology, Faculty of Science, Silpakorn University, Nakhon Pathom 7300, Thailand. 2Department of Chemistry, Faculty of Science Silpakorn University, Nakhon Pathom 7300, Thailand. ARTICLE INFO ABSTRACT Received on: 12/10/2019 Geldanamycin (1) was isolated as a major compound from Streptomyces zerumbet W14. It was then used as a precursor to Accepted on: 02/04/2020 synthesize two new geldanamycins: 17-(tryptamine)-17-demethoxygeldanamycin (2) and 17-(5′-methoxytryptamine)- Available online: 05/06/2020 17-demethoxygeldanamycin (3). The cytotoxicity activity of these two new compounds was evaluated and compared with the cytotoxicity of compound 1. Cytotoxicity activity was evaluated against a normal cell line, and three cancer cell lines using an 3-(4, 5-Dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) colorimetric assay. The Key words: solubility of these compounds was also determined. The solubility of compounds 2 and 3 in water was 290.69 Anticancer activity, cancer and 348.18 µM, higher than that of compound 1 by about 1.91 and 2.29 times, respectively. Compounds 2 and 3 cell lines, cytotoxicity showed moderate cytotoxic activity on Vero and human cervical carcinoma cells with IC50 values of >200.00 µg/ activity, solubility, ml. The strongest cytotoxicity of compound 3 was observed in human breast carcinoma cells (MCF-7) and human tryptamine-geldanamycin hepatocellular carcinoma cell line (HepG2) cells with IC50 values of 82.50 and 114.35 µg/ml, respectively, while the hybrids.
    [Show full text]