Cellular transformation is accompanied by extensive rewiring of many biological processes leading to augmented levels of distinct types of cellular stress, including proteotoxic stress. Cancer cells critically depend on stress-relief pathways for their survival. However, the mechanisms underlying the transcriptional initiation and maintenance of the oncogenic stress response remain elusive. Here, we show that the expression of heat shock transcription factor 1 (HSF1) and the downstream mediators of the heat shock response is transcriptionally upregulated in T cell acute lymphoblastic leukemia (T-ALL). Hsf1 ablation suppresses the growth of human T-ALL and eradicates leukemia in mouse models of T-ALL, while sparing normal hematopoiesis. HSF1 drives a compact transcriptional program and among the direct HSF1 targets, specific chaperones and co-chaperones mediate its critical role in T-ALL. Notably, we demonstrate that the central T-ALL oncogene NOTCH1 hijacks the cellular stress response machinery by inducing the expression of HSF1 and its downstream effectors. The NOTCH1 signaling status controls the levels of chaperone/co-chaperone complexes and predicts the response of T-ALL patient samples to HSP90 inhibition. Our data demonstrate an integral crosstalk between mediators of oncogene and non-oncogene addiction and reveal critical nodes of the heat shock response pathway that can be targeted therapeutically.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.


All prices are NET prices.

Additional information

Publisher’s note: Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.


  1. 1.

    Dai, C., Whitesell, L., Rogers, A. B. & Lindquist, S. Heat shock factor 1 is a powerful multifaceted modifier of carcinogenesis. Cell 130, 1005–1018 (2007).

  2. 2.

    Mendillo, M. L. et al. HSF1 drives a transcriptional program distinct from heat shock to support highly malignant human cancers. Cell 150, 549–562 (2012).

  3. 3.

    Santagata, S. et al. Tight coordination of protein translation and HSF1 activation supports the anabolic malignant state. Science 341, 1238303 (2013).

  4. 4.

    Xi, C., Hu, Y., Buckhaults, P., Moskophidis, D. & Mivechi, N. F. Heat shock factor Hsf1 cooperates with ErbB2 (Her2/Neu) protein to promote mammary tumorigenesis and metastasis. J. Biol. Chem. 287, 35646–35657 (2012).

  5. 5.

    Akerfelt, M., Morimoto, R. I. & Sistonen, L. Heat shock factors: integrators of cell stress, development and lifespan. Nat. Rev. Mol. Cell Biol. 11, 545–555 (2010).

  6. 6.

    Dai, C. The heat-shock, or HSF1-mediated proteotoxic stress, response in cancer: from proteomic stability to oncogenesis. Phil. Trans. R. Soc. B 373, 20160525 (2018).

  7. 7.

    Gomez-Pastor, R. et al. Abnormal degradation of the neuronal stress-protective transcription factor HSF1 in Huntington’s disease. Nat. Commun. 8, 14405 (2017).

  8. 8.

    Li, J., Labbadia, J. & Morimoto, R. I. Rethinking HSF1 in stress, development, and organismal health. Trends Cell Biol. (2017).

  9. 9.

    Luo, J., Solimini, N. L. & Elledge, S. J. Principles of cancer therapy: oncogene and non-oncogene addiction. Cell 136, 823–837 (2009).

  10. 10.

    Nagel, R., Semenova, E. A. & Berns, A. Drugging the addict: non-oncogene addiction as a target for cancer therapy. EMBO Rep. 17, 1516–1531 (2016).

  11. 11.

    Gomez-Pastor, R., Burchfiel, E. T. & Thiele, D. J. Regulation of heat shock transcription factors and their roles in physiology and disease. Nat. Rev. Mol. Cell Biol. 19, 4–19 (2018).

  12. 12.

    Whitesell, L. & Lindquist, S. Inhibiting the transcription factor HSF1 as an anticancer strategy. Expert Opin. Ther. Targets 13, 469–478 (2009).

  13. 13.

    Belver, L. & Ferrando, A. The genetics and mechanisms of T cell acute lymphoblastic leukaemia. Nat. Rev. Cancer 16, 494–507 (2016).

  14. 14.

    Palomero, T. et al. NOTCH1 directly regulates c-MYC and activates a feed-forward-loop transcriptional network promoting leukemic cell growth. Proc. Natl. Acad. Sci. USA 103, 18261–18266 (2006).

  15. 15.

    Weng, A. P. et al. c-Myc is an important direct target of Notch1 in T-cell acute lymphoblastic leukemia/lymphoma. Genes Dev. 20, 2096–2109 (2006).

  16. 16.

    Chou, S. D., Prince, T., Gong, J. & Calderwood, S. K. mTOR is essential for the proteotoxic stress response, HSF1 activation and heat shock protein synthesis. PloS One 7, e39679 (2012).

  17. 17.

    Cotto, J. J., Kline, M. & Morimoto, R. I. Activation of heat shock factor 1 DNA binding precedes stress-induced serine phosphorylation. Evidence for a multistep pathway of regulation. J. Biol. Chem. 271, 3355–3358 (1996).

  18. 18.

    Hietakangas, V. et al. PDSM, a motif for phosphorylation-dependent SUMO modification. Proc. Natl. Acad. Sci. USA 103, 45–50 (2006).

  19. 19.

    Holmberg, C. I. et al. Phosphorylation of serine 230 promotes inducible transcriptional activity of heat shock factor 1. EMBO J. 20, 3800–3810 (2001).

  20. 20.

    Hu, Y. & Mivechi, N. F. Promotion of heat shock factor Hsf1 degradation via adaptor protein filamin A-interacting protein 1-like (FILIP-1L). J. Biol. Chem. 286, 31397–31408 (2011).

  21. 21.

    Kourtis, N. et al. FBXW7 modulates cellular stress response and metastatic potential through HSF1 post-translational modification. Nat. Cell Biol. 17, 322–332 (2015).

  22. 22.

    Tang, Z. et al. MEK guards proteome stability and inhibits tumor-suppressive amyloidogenesis via HSF1. Cell 160, 729–744 (2015).

  23. 23.

    Westerheide, S. D., Anckar, J., Stevens, S. M. Jr, Sistonen, L. & Morimoto, R. I. Stress-inducible regulation of heat shock factor 1 by the deacetylase SIRT1. Science 323, 1063–1066 (2009).

  24. 24.

    Van Vlierberghe, P. et al. ETV6 mutations in early immature human T cell leukemias. J. Exp. Med. 208, 2571–2579 (2011).

  25. 25.

    Guettouche, T., Boellmann, F., Lane, W. S. & Voellmy, R. Analysis of phosphorylation of human heat shock factor 1 in cells experiencing a stress. BMC Biochem. 6, 4 (2005).

  26. 26.

    Palomero, T. et al. CUTLL1, a novel human T-cell lymphoma cell line with t(7;9) rearrangement, aberrant NOTCH1 activation and high sensitivity to gamma-secretase inhibitors. Leukemia 20, 1279–1287 (2006).

  27. 27.

    Brandvold, K. R. & Morimoto, R. I. The chemical biology of molecular chaperones—implications for modulation of proteostasis. J. Mol. Biol. 427, 2931–2947 (2015).

  28. 28.

    Aster, J. C. et al. Oncogenic forms of NOTCH1 lacking either the primary binding site for RBP-Jkappa or nuclear localization sequences retain the ability to associate with RBP-Jkappa and activate transcription. J. Biol. Chem. 272, 11336–11343 (1997).

  29. 29.

    King, B. et al. The ubiquitin ligase FBXW7 modulates leukemia-initiating cell activity by regulating MYC stability. Cell 153, 1552–1566 (2013).

  30. 30.

    O’Neil, J. et al. Activating Notch1 mutations in mouse models of T-ALL. Blood 107, 781–785 (2006).

  31. 31.

    Roderick, J. E. et al. c-Myc inhibition prevents leukemia initiation in mice and impairs the growth of relapsed and induction failure pediatric T-ALL cells. Blood 123, 1040–1050 (2014).

  32. 32.

    Xiao, X. et al. HSF1 is required for extra-embryonic development, postnatal growth and protection during inflammatory responses in mice. EMBO J. 18, 5943–5952 (1999).

  33. 33.

    Xiao, H. & Lis, J. T. Germline transformation used to define key features of heat-shock response elements. Science 239, 1139–1142 (1988).

  34. 34.

    Scherz-Shouval, R. et al. The reprogramming of tumor stroma by HSF1 is a potent enabler of malignancy. Cell 158, 564–578 (2014).

  35. 35.

    Taipale, M. et al. A quantitative chaperone interaction network reveals the architecture of cellular protein homeostasis pathways. Cell 158, 434–448 (2014).

  36. 36.

    Grabher, C., von Boehmer, H. & Look, A. T. Notch 1 activation in the molecular pathogenesis of T-cell acute lymphoblastic leukaemia. Nat. Rev. Cancer 6, 347–359 (2006).

  37. 37.

    Weng, A. P. et al. Activating mutations of NOTCH1 in human T cell acute lymphoblastic leukemia. Science 306, 269–271 (2004).

  38. 38.

    Wang, H. et al. Genome-wide analysis reveals conserved and divergent features of Notch1/RBPJ binding in human and murine T-lymphoblastic leukemia cells. Proc. Natl. Acad. Sci. USA 108, 14908–14913 (2011).

  39. 39.

    Mahat, D. B., Salamanca, H. H., Duarte, F. M., Danko, C. G. & Lis, J. T. Mammalian heat shock response and mechanisms underlying its genome-wide transcriptional regulation. Mol. Cell 62, 63–78 (2016).

  40. 40.

    Solis, E. J. et al. Defining the essential function of yeast Hsf1 reveals a compact transcriptional program for maintaining eukaryotic proteostasis. Mol. Cell 63, 60–71 (2016).

  41. 41.

    Kopan, R. & Ilagan, M. X. The canonical Notch signaling pathway: unfolding the activation mechanism. Cell 137, 216–233 (2009).

  42. 42.

    Ben-Bassat, H., Shlomai, Z., Kohn, G. & Prokocimer, M. Establishment of a human T-acute lymphoblastic leukemia cell line with a (16;20) chromosome translocation. Cancer Genet. Cytogenet. 49, 241–248 (1990).

  43. 43.

    Holmes, R. & Zuniga-Pflucker, J. C. The OP9-DL1 system: generation of T-lymphocytes from embryonic or hematopoietic stem cells in vitro. Cold Spring Harb. Protoc. 2009, pdbprot5156 (2009).

  44. 44.

    Core, L. J., Waterfall, J. J. & Lis, J. T. Nascent RNA sequencing reveals widespread pausing and divergent initiation at human promoters. Science 322, 1845–1848 (2008).

  45. 45.

    Yatim, A. et al. NOTCH1 nuclear interactome reveals key regulators of its transcriptional activity and oncogenic function. Mol. Cell 48, 445–458 (2012).

  46. 46.

    de Jonge, H. J. et al. Gene expression profiling in the leukemic stem cell-enriched CD34+ fraction identifies target genes that predict prognosis in normal karyotype AML. Leukemia 25, 1825–1833 (2011).

  47. 47.

    Fabbri, G. et al. Analysis of the chronic lymphocytic leukemia coding genome: role of NOTCH1 mutational activation. J. Exp. Med. 208, 1389–1401 (2011).

  48. 48.

    Rossi, D. et al. Mutations of NOTCH1 are an independent predictor of survival in chronic lymphocytic leukemia. Blood 119, 521–529 (2012).

  49. 49.

    Klinakis, A. et al. A novel tumour-suppressor function for the Notch pathway in myeloid leukaemia. Nature 473, 230–233 (2011).

  50. 50.

    Fabbri, G. et al. Common nonmutational NOTCH1 activation in chronic lymphocytic leukemia. Proc. Natl. Acad. Sci. USA 114, E2911–E2919 (2017).

  51. 51.

    Kamal, A. et al. A high-affinity conformation of Hsp90 confers tumour selectivity on Hsp90 inhibitors. Nature 425, 407–410 (2003).

  52. 52.

    Rodina, A. et al. The epichaperome is an integrated chaperome network that facilitates tumour survival. Nature 538, 397–401 (2016).

  53. 53.

    Akahane, K. et al. HSP90 inhibition leads to degradation of the TYK2 kinase and apoptotic cell death in T-cell acute lymphoblastic leukemia. Leukemia 30, 219–228 (2016).

  54. 54.

    Shrestha, L., Patel, H. J. & Chiosis, G. Chemical tools to investigate mechanisms associated with HSP90 and HSP70 in disease. Cell Chem. Biol. 23, 158–172 (2016).

  55. 55.

    Moulick, K. et al. Affinity-based proteomics reveal cancer-specific networks coordinated by Hsp90. Nat. Chem. Biol. 7, 818–826 (2011).

  56. 56.

    Taldone, T. et al. Synthesis of purine-scaffold fluorescent probes for heat shock protein 90 with use in flow cytometry and fluorescence microscopy. Bioorg. Med. Chem. Lett. 21, 5347–5352 (2011).

  57. 57.

    Herranz, D. et al. Metabolic reprogramming induces resistance to anti-NOTCH1 therapies in T cell acute lymphoblastic leukemia. Nat. Med. 21, 1182–1189 (2015).

  58. 58.

    Wang, Z. et al. Stabilization of Notch1 by the Hsp90 chaperone is crucial for T-cell leukemogenesis. Clin. Cancer Res. 23, 3834–3846 (2017).

  59. 59.

    Dai, C. & Sampson, S. B. HSF1: guardian of proteostasis in cancer. Trends Cell Biol. 26, 17–28 (2016).

  60. 60.

    Schopf, F. H., Biebl, M. M. & Buchner, J. The HSP90 chaperone machinery. Nat. Rev. Mol. Cell Biol. 18, 345–360 (2017).

  61. 61.

    Le Masson, F. et al. Identification of heat shock factor 1 molecular and cellular targets during embryonic and adult female meiosis. Mol. Cell. Biol. 31, 3410–3423 (2011).

  62. 62.

    Tatarek, J. et al. Notch1 inhibition targets the leukemia-initiating cells in a Tal1/Lmo2 mouse model of T-ALL. Blood 118, 1579–1590 (2011).

  63. 63.

    Taldone, T. et al. Heat shock protein 70 inhibitors. 2. 2,5'-thiodipyrimidines, 5-(phenylthio)pyrimidines, 2-(pyridin-3-ylthio)pyrimidines, and 3-(phenylthio)pyridines as reversible binders to an allosteric site on heat shock protein 70. J. Med. Chem. 57, 1208–1224 (2014).

  64. 64.

    Ntziachristos, P. et al. Genetic inactivation of the polycomb repressive complex 2 in T cell acute lymphoblastic leukemia. Nat. Med. 18, 298–301 (2012).

  65. 65.

    Jensen, L. J. et al. STRING 8—global view on proteins and their functional interactions in 630 organisms. Nucleic Acids Res. 37, D412–D416 (2009).

  66. 66.

    Gong, Y. et al. lncRNA-screen: an interactive platform for computationally screening long non-coding RNAs in large genomics datasets. BMC Genomics 18, 434 (2017).

  67. 67.

    Dobin, A. et al. STAR: ultrafast universal RNA-seq aligner. Bioinformatics 29, 15–21 (2013).

  68. 68.

    Love, M. I., Huber, W. & Anders, S. Moderated estimation of fold change and dispersion for RNA-seq data with DESeq2. Genome Biol. 15, 550 (2014).

  69. 69.

    Langmead, B. & Salzberg, S. L. Fast gapped-read alignment with Bowtie 2. Nat. Methods 9, 357–359 (2012).

  70. 70.

    Heinz, S. et al. Simple combinations of lineage-determining transcription factors prime cis-regulatory elements required for macrophage and B cell identities. Mol. Cell 38, 576–589 (2010).

  71. 71.

    Huber, W. et al. Orchestrating high-throughput genomic analysis with Bioconductor. Nat. Methods 12, 115–121 (2015).

  72. 72.

    Tsirigos, A., Haiminen, N., Bilal, E. & Utro, F. GenomicTools: a computational platform for developing high-throughput analytics in genomics. Bioinformatics 28, 282–283 (2012).

  73. 73.

    Wickham, H. Ggplot2: Elegant Graphics for Data Analysis (Springer, New York, 2009).

  74. 74.

    Gautier, L., Cope, L., Bolstad, B. M. & Irizarry, R. A. affy—analysis of Affymetrix GeneChip data at the probe level. Bioinformatics 20, 307–315 (2004).

  75. 75.

    Zhou, X. et al. The Human Epigenome Browser at Washington University. Nat. Methods 8, 989–990 (2011).

  76. 76.

    Grant, C. E., Bailey, T. L. & Noble, W. S. FIMO: scanning for occurrences of a given motif. Bioinformatics 27, 1017–1018 (2011).

  77. 77.

    Wang, M., Zhao, Y. & Zhang, B. Efficient test and visualization of multi-set intersections. Sci. Rep. 5, 16923 (2015).

  78. 78.

    Huang da, W., Sherman, B. T. & Lempicki, R. A. Systematic and integrative analysis of large gene lists using DAVID bioinformatics resources. Nat. Protoc. 4, 44–57 (2009).

  79. 79.

    Rahl, P. B. et al. c-Myc regulates transcriptional pause release. Cell 141, 432–445 (2010).

  80. 80.

    Wang, H. et al. NOTCH1-RBPJ complexes drive target gene expression through dynamic interactions with superenhancers. Proc. Natl. Acad. Sci. USA 111, 705–710 (2014).

Download references


We thank all members of the Aifantis laboratory for discussions throughout the duration of this project; T. Papagiannakopoulos and P. Ntziachristos for critical assessment of this work; E. Christians (UPMC Univ. Paris 06, CNRS) for the Hsf1f/f mice; A. Heguy and the NYU Genome Technology Center (supported in part by National Institutes of Health (NIH)/National Cancer Institute (NCI) grant P30CA016087-30) for expertise with sequencing experiments; the NYU Histology Core (5P30CA16087-31) for assistance; C. Loomis and L. Chiriboga for immunohistochemistry experiments; C. Jamieson (UCSD) for human LICs; The ECOG-ACRIN Cancer Research Group for clinical specimens. This work has used computing resources at the High Performance Computing Facility at the NYU Medical Center. A.T. is supported by a Research Scholar Grant (RSG-15-189-01-RMC) from the American Cancer Society and a Leukemia & Lymphoma Society New Idea Award (8007-17). I.A. is supported by the NIH (R01CA133379, R01CA105129, R01CA149655, 5R01CA173636, 1R01CA194923), the NYSTEM program of the New York State Health Department (NYSTEM-N11G-255) and the Leukemia & Lymphoma Society (LLS) Translational Research Program (TRP). J.C.B. was supported by CONACyT​ (​​FOSISSS 2015-1-261848​)​ and​ ​IMSS​ (FIS/IMSS/PROT/G14/1289​). J.M. was supported by KWF BUIT2012-5358. N.K. is supported by a Human Frontiers Science Program (HFSP) Long Term Fellowship (LT000150/2013-L) and previously by a Charles H. Revson Senior Fellowship in Biomedical Science (15–31) and a European Molecular Biology Organization (EMBO) Long Term Fellowship (ALTF 850-2012).

Author information

Author notes

    • Jasper Mullenders

    Present address: Hubrecht Institute, Royal Netherlands Academy of Arts and Sciences (KNAW) and University Medical Centre (UMC), Utrecht, the Netherlands

    • Alberto Ambesi-Impiombato

    Present address: PsychoGenics Inc., Tarrytown, New York, NY, USA

  1. These authors contributed equally: Nikos Kourtis, Charalampos Lazaris.


  1. Department of Pathology and Laura and Isaac Perlmutter Cancer Center, NYU School of Medicine, New York, NY, USA

    • Nikos Kourtis
    • , Charalampos Lazaris
    • , Kathryn Hockemeyer
    • , Jasper Mullenders
    • , Yixiao Gong
    • , Thomas Trimarchi
    • , Kamala Bhatt
    • , Hai Hu
    • , Aristotelis Tsirigos
    •  & Iannis Aifantis
  2. Molecular Biomedicine Program, CINVESTAV IPN, Mexico City, Mexico

    • Juan Carlos Balandrán
  3. CONACYT–Centro de Investigacion Biomedica de Oriente, IMSS Delegacion Puebla, Atlixco, Mexico

    • Juan Carlos Balandrán
  4. Haematology and Medical Oncology, Department of Medicine, Weill Cornell Medical College, New York, NY, USA

    • Juan Carlos Balandrán
    • , Alejandra R. Jimenez
    •  & Monica L. Guzman
  5. Program in Chemical Biology, Sloan Kettering Institute, New York, NY, USA

    • Liza Shrestha
    •  & Gabriela Chiosis
  6. Institute for Cancer Genetics, Department of Pathology and Department of Pediatrics, Columbia University, New York, NY, USA

    • Alberto Ambesi-Impiombato
    •  & Adolfo A. Ferrando
  7. Department of Molecular, Cell and Cancer Biology, University of Massachusetts Medical School, Worcester, MA, USA

    • Michelle Kelliher
  8. Montefiore Medical Center, New York, NY, USA

    • Elisabeth Paietta
  9. Applied Bioinformatics Laboratories, NYU School of Medicine, New York, NY, USA

    • Aristotelis Tsirigos


  1. Search for Nikos Kourtis in:

  2. Search for Charalampos Lazaris in:

  3. Search for Kathryn Hockemeyer in:

  4. Search for Juan Carlos Balandrán in:

  5. Search for Alejandra R. Jimenez in:

  6. Search for Jasper Mullenders in:

  7. Search for Yixiao Gong in:

  8. Search for Thomas Trimarchi in:

  9. Search for Kamala Bhatt in:

  10. Search for Hai Hu in:

  11. Search for Liza Shrestha in:

  12. Search for Alberto Ambesi-Impiombato in:

  13. Search for Michelle Kelliher in:

  14. Search for Elisabeth Paietta in:

  15. Search for Gabriela Chiosis in:

  16. Search for Monica L. Guzman in:

  17. Search for Adolfo A. Ferrando in:

  18. Search for Aristotelis Tsirigos in:

  19. Search for Iannis Aifantis in:


N.K. and I.A. designed the experiments and wrote the manuscript. N.K. performed most of the experiments. C.L. designed and performed bioinformatics analysis of genome-wide data. K.H., J.C.B, A.R.J., J.M. and T.T. performed experiments. Y.G. performed RNA-seq analysis. K.B. and H.H maintained the mouse colonies. L.S. provided molecular chaperones inhibitors. M.K. provided Tal1 cells. A.A.-I., E.P. and A.A.F. provided T-ALL patient expression data and patient samples. G.C. and M.L.G. provided molecular chaperones inhibitors and ideas. A.T. designed and performed bioinformatics analysis of genome-wide data and contributed ideas.

Competing interests

The authors declare no competing interests.

Corresponding authors

Correspondence to Nikos Kourtis or Iannis Aifantis.

Supplementary Information

  1. Supplementary Text and Figures

    Supplementary Figures 1–11 and Supplementary Tables 1 and 3

  2. Reporting Summary

  3. Supplementary Table 2

    Notch1-IC-interacting proteins in the mouse T-ALL 720 cell line or 293T cells

About this article

Publication history




Issue Date