Skip to main content

Thank you for visiting You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

Targeting unfolded protein response using albumin-encapsulated nanoparticles attenuates temozolomide resistance in glioblastoma



Chemoresistant cancer cells frequently exhibit a state of chronically activated endoplasmic reticulum (ER) stress. Engaged with ER stress, the unfolded protein response (UPR) is an adaptive reaction initiated by the accumulation of misfolded proteins. Protein disulfide isomerase (PDI) is a molecular chaperone known to be highly expressed in glioblastomas with acquired resistance to temozolomide (TMZ). We investigate whether therapeutic targeting of PDI provides a rationale to overcome chemoresistance.


The activity of PDI was suppressed in glioblastoma cells using a small molecule inhibitor CCF642. Either single or combination treatment with TMZ was used. We prepared nanoformulation of CCF642 loaded in albumin as a drug carrier for orthotopic tumour model.


Inhibition of PDI significantly enhances the cytotoxic effect of TMZ on glioblastoma cells. More importantly, inhibition of PDI is able to sensitise glioblastoma cells that are initially resistant to TMZ treatment. Nanoformulation of CCF642 is well-tolerated and effective in suppressing tumour growth. It activates cell death-triggering UPR beyond repair and induces ER perturbations through the downregulation of PERK signalling. Combination treatment of TMZ with CCF642 significantly reduces tumour growth compared with either modality alone.


Our study demonstrates modulation of ER stress by targeting PDI as a promising therapeutic rationale to overcome chemoresistance.

This is a preview of subscription content, access via your institution

Access options

Rent or buy this article

Get just this article for as long as you need it


Prices may be subject to local taxes which are calculated during checkout

Fig. 1: Inhibition of PDI attenuates TMZ resistance in vitro.
Fig. 2: Inhibition of PDI-sensitised glioblastoma cells upon TMZ treatment via UPR-mediated cell death in vitro.
Fig. 3: Characterisation of HSA-encapsulated CCF642 nanoparticles.
Fig. 4: PDI inhibition suppresses glioblastoma growth and attenuates TMZ resistance in vivo.
Fig. 5: Schematic diagram of the molecular mechanism underlying chronic ER stress-induced TMZ resistance.

Data availability

The data presented in this study are available from the corresponding author upon reasonable request.


  1. Saunders NA, Simpson F, Thompson EW, Hill MM, Endo-Munoz L, Leggatt G, et al. Role of intratumoural heterogeneity in cancer drug resistance: molecular and clinical perspectives. EMBO Mol Med. 2012;4:675–84.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  2. Happold C, Roth P, Wick W, Schmidt N, Florea AM, Silginer M, et al. Distinct molecular mechanisms of acquired resistance to temozolomide in glioblastoma cells. J Neurochem. 2012;122:444–55.

    Article  CAS  PubMed  Google Scholar 

  3. Ron D, Walter P. Signal integration in the endoplasmic reticulum unfolded protein response. Nat Rev Mol Cell Biol. 2007;8:519–29.

    Article  CAS  PubMed  Google Scholar 

  4. Salaroglio IC, Panada E, Moiso E, Buondonno I, Provero P, Rubinstein M, et al. PERK induces resistance to cell death elicited by endoplasmic reticulum stress and chemotherapy. Mol Cancer. 2017;16:91.

    Article  PubMed  PubMed Central  Google Scholar 

  5. Sun S, Lee D, Ho AS, Pu JK, Zhang XQ, Lee NP, et al. Inhibition of prolyl 4-hydroxylase, beta polypeptide (P4HB) attenuates temozolomide resistance in malignant glioma via the endoplasmic reticulum stress response (ERSR) pathways. Neuro Oncol. 2013;15:562–77.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Sun S, Wong TS, Zhang XQ, Pu JK, Lee NP, Day PJ, et al. Protein alterations associated with temozolomide resistance in subclones of human glioblastoma cell lines. J Neurooncol. 2012;107:89–100.

    Article  CAS  PubMed  Google Scholar 

  7. Sun S, Kiang KMY, Ho ASW, Lee D, Poon MW, Xu FF, et al. Endoplasmic reticulum chaperone prolyl 4-hydroxylase, beta polypeptide (P4HB) promotes malignant phenotypes in glioma via MAPK signaling. Oncotarget. 2017;8:71911–23.

    Article  PubMed  PubMed Central  Google Scholar 

  8. Sun S, Kiang KM, Leung GK. Chaperone protein P4HB predicts temozolomide response and prognosis in malignant glioma. Oncol Lett. 2022;24:264.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Samanta S, Tamura S, Dubeau L, Mhawech-Fauceglia P, Miyagi Y, Kato H, et al. Expression of protein disulfide isomerase family members correlates with tumor progression and patient survival in ovarian cancer. Oncotarget. 2017;8:103543–56.

    Article  PubMed  PubMed Central  Google Scholar 

  10. Yu SJ, Won JK, Ryu HS, Choi WM, Cho H, Cho EJ, et al. A novel prognostic factor for hepatocellular carcinoma: protein disulfide isomerase. Korean J Intern Med. 2014;29:580–7.

    Article  PubMed  PubMed Central  Google Scholar 

  11. Kim KM, An AR, Park HS, Jang KY, Moon WS, Kang MJ, et al. Combined expression of protein disulfide isomerase and endoplasmic reticulum oxidoreductin 1-alpha is a poor prognostic marker for non-small cell lung cancer. Oncol Lett. 2018;16:5753–60.

    CAS  PubMed  PubMed Central  Google Scholar 

  12. Goplen D, Wang J, Enger PO, Tysnes BB, Terzis AJ, Laerum OD, et al. Protein disulfide isomerase expression is related to the invasive properties of malignant glioma. Cancer Res. 2006;66:9895–902.

    Article  CAS  PubMed  Google Scholar 

  13. Lovat PE, Corazzari M, Armstrong JL, Martin S, Pagliarini V, Hill D, et al. Increasing melanoma cell death using inhibitors of protein disulfide isomerases to abrogate survival responses to endoplasmic reticulum stress. Cancer Res. 2008;68:5363–9.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Eirich J, Braig S, Schyschka L, Servatius P, Hoffmann J, Hecht S, et al. A small molecule inhibits protein disulfide isomerase and triggers the chemosensitization of cancer cells. Angew Chem Int Ed Engl. 2014;53:12960–5.

    Article  CAS  PubMed  Google Scholar 

  15. Kaplan A, Gaschler MM, Dunn DE, Colligan R, Brown LM, Palmer AG 3rd, et al. Small molecule-induced oxidation of protein disulfide isomerase is neuroprotective. Proc Natl Acad Sci USA. 2015;112:E2245–52.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Vatolin S, Phillips JG, Jha BK, Govindgari S, Hu J, Grabowski D, et al. Novel protein disulfide isomerase inhibitor with anticancer activity in multiple myeloma. Cancer Res. 2016;76:3340–50.

    Article  CAS  PubMed  Google Scholar 

  17. Liu Y, Ji W, Shergalis A, Xu J, Delaney AM, Calcaterra A, et al. Activation of the unfolded protein response via inhibition of protein disulfide isomerase decreases the capacity for DNA repair to sensitize glioblastoma to radiotherapy. Cancer Res. 2019;79:2923–32.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Xu S, Liu Y, Yang K, Wang H, Shergalis A, Kyani A, et al. Inhibition of protein disulfide isomerase in glioblastoma causes marked downregulation of DNA repair and DNA damage response genes. Theranostics. 2019;9:2282–98.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Lampson LA. Monoclonal antibodies in neuro-oncology: getting past the blood-brain barrier. MAbs. 2011;3:153–60.

    Article  PubMed  PubMed Central  Google Scholar 

  20. Zhou G, Jin X, Zhu P, Yao JU, Zhang Y, Teng L, et al. Human serum albumin nanoparticles as a novel delivery system for cabazitaxel. Anticancer Res. 2016;36:1649–56.

    CAS  PubMed  Google Scholar 

  21. Haley B, Frenkel E. Nanoparticles for drug delivery in cancer treatment. Urol Oncol. 2008;26:57–64.

    Article  CAS  PubMed  Google Scholar 

  22. Zong Y, Wu J, Shen K. Nanoparticle albumin-bound paclitaxel as neoadjuvant chemotherapy of breast cancer: a systematic review and meta-analysis. Oncotarget. 2017;8:17360–72.

    Article  PubMed  PubMed Central  Google Scholar 

  23. McBain C, Lawrie TA, Rogozinska E, Kernohan A, Robinson T, Jefferies S. Treatment options for progression or recurrence of glioblastoma: a network meta-analysis. Cochrane Database Syst Rev. 2021;5:CD013579.

    PubMed  Google Scholar 

  24. Montane J, de Pablo S, Obach M, Cadavez L, Castano C, Alcarraz-Vizan G, et al. Protein disulfide isomerase ameliorates beta-cell dysfunction in pancreatic islets overexpressing human islet amyloid polypeptide. Mol Cell Endocrinol. 2016;420:57–65.

    Article  CAS  PubMed  Google Scholar 

  25. Hasipek M, Grabowski D, Guan Y, Alugubelli RR, Tiwari AD, Gu X, et al. Therapeutic targeting of protein disulfide isomerase PDIA1 in multiple myeloma. Cancers. 2021;13:2649.

  26. Sisinni L, Pietrafesa M, Lepore S, Maddalena F, Condelli V, Esposito F, et al. Endoplasmic reticulum stress and unfolded protein response in breast cancer: the balance between apoptosis and autophagy and its role in drug resistance. Int J Mol Sci. 2019;20:857.

  27. Luo B, Lee AS. The critical roles of endoplasmic reticulum chaperones and unfolded protein response in tumorigenesis and anticancer therapies. Oncogene. 2013;32:805–18.

    Article  CAS  PubMed  Google Scholar 

  28. Markouli M, Strepkos D, Papavassiliou AG, Piperi C. Targeting of endoplasmic reticulum (ER) stress in gliomas. Pharm Res. 2020;157:104823.

    Article  CAS  Google Scholar 

Download references


This work was supported by the Health and Medical Research Fund (HMRF), project no.08191986, of the Food and Health Bureau (Hong Kong). We would like to thank the University of Hong Kong LKS Faculty of Medicine the Centre of PanorOmic Sciences (Imaging and flow cytometry core), and the Electron Microscope Unit for their assistance and services.


This work was supported by the Health and Medical Research Fund (HMRF), project no. 08191986, of the Food and Health Bureau (Hong Kong).

Author information

Authors and Affiliations



KK performed the experiments, wrote the original draft and involved in conceptualisation and funding acquisition. WT, JL and TL performed the experiments and provided technical support. QS and HS helped in the preparation of nanoparticles and provided technical support. NL participated in the formal data analyses. ZZ and GL supervised the entire study and provided critical review on the manuscript. All authors reviewed the final manuscript.

Corresponding authors

Correspondence to Zhiyuan Zhu or Gilberto Ka-Kit Leung.

Ethics declarations

Competing interests

HCS is a scientific advisor of EN Technology Limited in which he owns some equity, and also a managing director of the research centre, namely Advanced Biomedical Instrumentation Centre Limited. The works in the paper are however not directly related to the works of these two entities, as far as we know. The remaining authors declare no competing interests.

Ethics approval and consent to participate

Animal work was performed according to guidelines approved by the Committee on the Use of Live Animal for Teaching and Research, The University of Hong Kong. The use of animals and all experimental procedures were conducted in compliance to ARRIVE guidelines.

Consent for publication

Not applicable.

Additional information

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

Rights and permissions

Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.

Reprints and Permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Kiang, K.MY., Tang, W., Song, Q. et al. Targeting unfolded protein response using albumin-encapsulated nanoparticles attenuates temozolomide resistance in glioblastoma. Br J Cancer 128, 1955–1963 (2023).

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI:


Quick links