Research Article

Depleting tumor-associated Tregs via nanoparticle-mediated hyperthermia to enhance anti-CTLA-4 immunotherapy

*Author for correspondence:

E-mail Address: hongweic@umich.edu

Department of Pharmaceutical Sciences, College of Pharmacy, University of Michigan, Ann Arbor, MI 48109, USA

^‡Authors contributed equally

Search for more papers by this author

Xin Luan^‡

Department of Pharmaceutical Sciences, College of Pharmacy, University of Michigan, Ann Arbor, MI 48109, USA

^‡Authors contributed equally

Search for more papers by this author

Hayley J Paholak

Department of Pharmaceutical Sciences, College of Pharmacy, University of Michigan, Ann Arbor, MI 48109, USA

Search for more papers by this author

Joseph P Burnett

Department of Pharmaceutical Sciences, College of Pharmacy, University of Michigan, Ann Arbor, MI 48109, USA

Search for more papers by this author

Nicholas O Stevers

Department of Pharmaceutical Sciences, College of Pharmacy, University of Michigan, Ann Arbor, MI 48109, USA

Search for more papers by this author

Kanokwan Sansanaphongpricha

Department of Pharmaceutical Sciences, College of Pharmacy, University of Michigan, Ann Arbor, MI 48109, USA

Current address: National Nanotechnology Center, National Science and Technology Development Agency, Pathum Thani, 12120, Thailand

Search for more papers by this author

Miao He

Department of Pharmaceutical Sciences, College of Pharmacy, University of Michigan, Ann Arbor, MI 48109, USA

Search for more papers by this author

Alfred E Chang

Department of Surgery, School of Medicine, University of Michigan, Ann Arbor, MI 48109, USA

Search for more papers by this author

Qiao Li

Department of Surgery, School of Medicine, University of Michigan, Ann Arbor, MI 48109, USA

Search for more papers by this author

Duxin Sun

**Author for correspondence:

E-mail Address: duxins@umich.edu

Department of Pharmaceutical Sciences, College of Pharmacy, University of Michigan, Ann Arbor, MI 48109, USA

Search for more papers by this author

Published Online:23 Dec 2019https://doi.org/10.2217/nnm-2019-0190

View article

Abstract

Aim: We aim to demonstrate that a local nanoparticle-mediated hyperthermia can effectively eliminate tumor-associated Tregs and thereby boost checkpoint blockade-based immunotherapy. Materials & methods: Photothermal therapy (PTT), mediated with systemically administered stealthy iron-oxide nanoparticles, was applied to treat BALB/c mice bearing 4T1 murine breast tumors. Flow cytometry was applied to evaluate both Treg and CD8⁺ T-cell population. Tumor growth following combination therapy of both PTT and anti-CTLA-4 was further evaluated. Results: Our data reveal that tumor-associated Tregs can be preferentially depleted via iron-oxide nanoparticles-mediated PTT. When combining PTT with anti-CTLA-4 immunotherapy, we demonstrate a significant inhibition of syngeneic 4T1 tumor growth. Conclusion: This study offers a novel strategy to overcome Treg-mediated immunosuppression and thereby to boost cancer immunotherapy.

Graphical abstract

Keywords:

Papers of special note have been highlighted as: • of interest; •• of considerable interest

References

1. Couzin-Frankel J . Cancer immunotherapy. Science 342(6165), 1432–1433 (2013).
Medline CASGoogle Scholar
2. Emens LA , Ascierto PA , Darcy PK et al. Cancer immunotherapy: opportunities and challenges in the rapidly evolving clinical landscape. Eur. J. Cancer 81, 116–129 (2017).
Medline CASGoogle Scholar
3. Zou WP , Wolchok JD , Chen LP . PD-L1 (B7-H1) and PD-1 pathway blockade for cancer therapy: mechanisms, response biomarkers, and combinations. Sci. Transl. Med. 8(328), 328rv4 (2016).
MedlineGoogle Scholar
4. Sharma P , Hu-Lieskovan S , Wargo JA , Ribas A . Primary, adaptive, and acquired resistance to cancer immunotherapy. Cell 168(4), 707–723 (2017).
Medline CASGoogle Scholar
5. Ott PA , Hodi FS , Kaufman HL , Wigginton JM , Wolchok JD . Combination immunotherapy: a road map. J. Immunother. Cancer 5, 16 (2017).
MedlineGoogle Scholar
6. Liu C , Workman CJ , Vignali DA . Targeting regulatory T cells in tumors. Febs J. 283(14), 2731–2748 (2016).
Medline CASGoogle Scholar
7. Tanaka A , Sakaguchi S . Regulatory T cells in cancer immunotherapy. Cell Res. 27(1), 109–118 (2017). •• A comprehensive review of Treg functions in tumor tissues and stragegies to target Tregs.
Medline CASGoogle Scholar
8. Piao YJ , Kim HS , Hwang EH , Woo J , Zhang M , Moon WK . Breast cancer cell-derived exosomes and macrophage polarization are associated with lymph node metastasis. Oncotarget 9(7), 7398–7410 (2018).
MedlineGoogle Scholar
9. Beyer M , Schultze JL . Regulatory T cells in cancer. Blood 108(3), 804–811 (2006).
Medline CASGoogle Scholar
10. Kavanagh B , O'Brien S , Lee D et al. CTLA4 blockade expands FoxP3(⁺) regulatory and activated effector CD4(⁺) T cells in a dose-dependent fashion. Blood 112(4), 1175–1183 (2008).
Medline CASGoogle Scholar
11. Chaudhary B , Elkord E . Regulatory T cells in the tumor microenvironment and cancer progression: role and therapeutic targeting. Vaccines (Basel) 4(3), E28 (2016).
MedlineGoogle Scholar
12. Maj T , Wang W , Crespo J et al. Oxidative stress controls regulatory T cell apoptosis and suppressor activity and PD-L1-blockade resistance in tumor. Nat. Immunol. 18(12), 1332–1341 (2017).
Medline CASGoogle Scholar
13. Chao JL , Savage PA . Unlocking the complexities of tumor-associated regulatory T cells. J. Immunol. 200(2), 415–421 (2018). • An interesting work of reviewing the advances in the understanding of tumor-associated Treg diversity.
Medline CASGoogle Scholar
14. Rech AJ , Mick R , Martin S et al. CD25 blockade depletes and selectively reprograms regulatory T cells in concert with immunotherapy in cancer patients. Sci. Transl. Med. 4(134), 134ra62 (2012).
MedlineGoogle Scholar
15. Jacobs JFM , Punt CJA , Lesterhuis WJ et al. Dendritic cell vaccination in combination with anti-CD25 monoclonal antibody treatment: a Phase I/II study in metastatic melanoma patients. Clin. Cancer Res. 16(20), 5067–5078 (2010).
Medline CASGoogle Scholar
16. Plitas G , Konopacki C , Wu K et al. Regulatory T cells exhibit distinct features in human breast cancer. Immunity 45(5), 1122–1134 (2016). •• An important work demonstrating the potent immunosuppressive features of the tumor-resident Tregs in human breast carcinomas.
Medline CASGoogle Scholar
17. De Simone M , Arrigoni A , Rossetti G et al. Transcriptional landscape of human tissue lymphocytes unveils uniqueness of tumor-infiltrating T regulatory cells. Immunity 45(5), 1135–1147 (2016).
MedlineGoogle Scholar
18. Vargas FA , Furness AJS , Litchfield K et al. Fc effector function contributes to the activity of human anti-CTLA-4 antibodies. Cancer Cell 33(4), 649–663 (2018). •• An interesting work demonstrating the Fc-gamma receptor-mediated intratumoral Treg depletion by anti-CTLA-4 antibodies.
MedlineGoogle Scholar
19. Selby MJ , Engelhardt JJ , Quigley M et al. Anti-CTLA-4 antibodies of IgG2a isotype enhance antitumor activity through reduction of intratumoral regulatory T cells. Cancer Immunol. Res. 1(1), 32–42 (2013).
Medline CASGoogle Scholar
20. Romano E , Kusio-Kobialka M , Foukas PG et al. Ipilimumab-dependent cell-mediated cytotoxicity of regulatory T cells ex vivo by nonclassical monocytes in melanoma patients. Proc. Natl Acad. Sci. USA 112(19), 6140–6145 (2015).
Medline CASGoogle Scholar
21. Sharma A , Subudhi SK , Blando J et al. Anti-CTLA-4 immunotherapy does not deplete FOXP3(⁺) regulatory T cells (Tregs) in human cancers. Clin. Cancer Res. 25(4), 1233–1238 (2019). • An interesting work of reporting that the anti-CTLA-4 anbibodies do not deplete Tregs in human tumors.
Medline CASGoogle Scholar
22. Postow MA , Chesney J , Pavlick AC et al. Nivolumab and ipilimumab versus ipilimumab in untreated melanoma. N. Engl. J. Med. 372(21), 2006–2017 (2015).
MedlineGoogle Scholar
23. Koumarianou A , Christodoulou MI , Patapis P et al. The effect of metronomic versus standard chemotherapy on the regulatory to effector T-cell equilibrium in cancer patients. Exp. Hematol. Oncol. 3(1), 3 (2014).
MedlineGoogle Scholar
24. Park SS , Dong H , Liu X et al. PD-1 restrains radiotherapy-induced abscopal effect. Cancer Immunol. Res. 3(6), 610–619 (2015).
Medline CASGoogle Scholar
25. Hou JJ , Greten TF , Xia Q . Immunosuppressive cell death in cancer. Nat. Rev. Immunol. 17(6), 401–401 (2017).
Medline CASGoogle Scholar
26. Li SY , Liu Y , Xu CF et al. Restoring anti-tumor functions of T cells via nanoparticle-mediated immune checkpoint modulation. J. Control. Rel. 231, 17–28 (2016).
Medline CASGoogle Scholar
27. Hussein EA , Zagho MM , Nasrallah GK , Elzatahry AA . Recent advances in functional nanostructures as cancer photothermal therapy. Int. J. Nanomed. 13, 2897–2906 (2018). • An important review of the progresses in nanoparticle-mediated photothermal cancer therapy.
Medline CASGoogle Scholar
28. Yang K , Hu LL , Ma XX et al. Multimodal imaging guided photothermal therapy using functionalized graphene nanosheets anchored with magnetic nanoparticles. Adv. Mater. 24(14), 1868–1872 (2012).
Medline CASGoogle Scholar
29. Guo LR , Yan DD , Yang DF et al. Combinatorial photothermal and immuno cancer therapy using chitosan-coated hollow copper sulfide nanoparticles. ACS Nano 8(6), 5670–5681 (2014).
Medline CASGoogle Scholar
30. Liang C , Diao S , Wang C et al. Tumor metastasis inhibition by imaging-guided photothermal therapy with single-walled carbon nanotubes. Adv. Mater. 26(32), 5646–5652 (2014).
Medline CASGoogle Scholar
31. Wyckoff JB , Wang Y , Lin EY et al. Direct visualization of macrophage-assisted tumor cell intravasation in mammary tumors. Cancer Res. 67(6), 2649–2656 (2007).
Medline CASGoogle Scholar
32. Chauhan VP , Jain RK . Strategies for advancing cancer nanomedicine. Nat. Mater. 12(11), 958–962 (2013).
Medline CASGoogle Scholar
33. Hansen AE , Petersen AL , Henriksen JR et al. Positron emission tomography based elucidation of the enhanced permeability and retention effect in dogs with cancer using copper-64 liposomes. ACS Nano 9(7), 6985–6995 (2015).
Medline CASGoogle Scholar
34. Toraya-Brown S , Sheen MR , Zhang P et al. Local hyperthermia treatment of tumors induces CD8(⁺) T cell-mediated resistance against distal and secondary tumors. Nanomedicine 10(6), 1273–1285 (2014).
Medline CASGoogle Scholar
35. Wang C , Xu L , Liang C , Xiang J , Peng R , Liu Z . Immunological responses triggered by photothermal therapy with carbon nanotubes in combination with anti-CTLA-4 therapy to inhibit cancer metastasis. Adv. Mater. 26(48), 8154–8162 (2014).
Medline CASGoogle Scholar
36. Chen Q , Xu L , Liang C , Wang C , Peng R , Liu Z . Photothermal therapy with immune-adjuvant nanoparticles together with checkpoint blockade for effective cancer immunotherapy. Nat. Commun. 7, 13193 (2016).
Medline CASGoogle Scholar
37. Cano-Mejia J , Burga RA , Sweeney EE et al. Prussian blue nanoparticle-based photothermal therapy combined with checkpoint inhibition for photothermal immunotherapy of neuroblastoma. Nanomed. Nanotechnol. 13(2), 771–781 (2017).
Medline CASGoogle Scholar
38. Chen HW , Burnett J , Zhang FX , Zhang JM , Paholak H , Sun DX . Highly crystallized iron oxide nanoparticles as effective and biodegradable mediators for photothermal cancer therapy. J. Mater. Chem. B 2(7), 757–765 (2014).
Medline CASGoogle Scholar
39. Zhou ZG , Sun YA , Shen JC et al. Iron/iron oxide core/shell nanoparticles for magnetic targeting MRI and near-infrared photothermal therapy. Biomaterials 35(26), 7470–7478 (2014).
Medline CASGoogle Scholar
40. Chen HW , Wang LY , Yeh J et al. Reducing non-specific binding and uptake of nanoparticles and improving cell targeting with an antifouling PEO-b-P γ MPS copolymer coating. Biomaterials 31(20), 5397–5407 (2010).
Medline CASGoogle Scholar
41. Paholak HJ , Stevers NO , Chen HW et al. Elimination of epithelial-like and mesenchymal-like breast cancer stem cells to inhibit metastasis following nanoparticle-mediated photothermal therapy. Biomaterials 104, 145–157 (2016).
Medline CASGoogle Scholar
42. Yao X , Ahmadzadeh M , Lu YC et al. Levels of peripheral CD4(⁺)FoxP3(⁺) regulatory T cells are negatively associated with clinical response to adoptive immunotherapy of human cancer. Blood 119(24), 5688–5696 (2012). •• Reports the clinical correlation between the density of circulating Tregs and the therapeutic outcomes following adoptive immunotherapy.
Medline CASGoogle Scholar
43. Hirsch LR , Stafford RJ , Bankson JA et al. Nanoshell-mediated near-infrared thermal therapy of tumors under magnetic resonance guidance. Proc. Natl Acad. Sci. USA 100(23), 13549–13554 (2003).
Medline CASGoogle Scholar
44. Cabral H , Matsumoto Y , Mizuno K et al. Accumulation of sub-100 nm polymeric micelles in poorly permeable tumours depends on size. Nat. Nanotechnol. 6(12), 815–823 (2011).
Medline CASGoogle Scholar
45. Gu Y , Liu YF , Fu L et al. Tumor-educated B cells selectively promote breast cancer lymph node metastasis by HSPA4-targeting IgG. Nat. Med. 25(2), 312–322 (2019).
Medline CASGoogle Scholar
46. Plitas G , Konopacki C , Wu KM et al. Regulatory T cells exhibit distinct features in human breast cancer. Immunity 45(5), 1122–1134 (2016).
Medline CASGoogle Scholar
47. Liu ZQ , Kim JH , Falo LD , You ZY . Tumor regulatory T cells potently abrogate antitumor immunity. J. Immunol. 182(10), 6160–6167 (2009).
Medline CASGoogle Scholar
48. Goudin N , Chappert P , Megret J , Gross DA , Rocha B , Azogui O . Depletion of regulatory T cells induces high numbers of dendritic cells and unmasks a subset of anti-tumour CD8(⁺)CD11c(⁺)PD-1(lo) effector T cells. PLoS ONE 11(6), e0157822 (2016).
MedlineGoogle Scholar
49. Yi JS , Cox MA , Zajac AJ . T-cell exhaustion: characteristics, causes and conversion. Immunology 129(4), 474–481 (2010).
Medline CASGoogle Scholar
50. Crespo J , Sun HY , Welling TH , Tian ZG , Zou WP . T cell anergy, exhaustion, senescence, and stemness in the tumor microenvironment. Curr. Opin. Immunol. 25(2), 214–221 (2013).
Medline CASGoogle Scholar
51. Tao K , Fang M , Alroy J , Sahagian GG . Imagable 4T1 model for the study of late stage breast cancer. BMC Cancer 8, 228 (2008).
MedlineGoogle Scholar
52. Melancon MP , Zhou M , Li C . Cancer theranostics with near-infrared light-activatable multimodal nanoparticles. Acc. Chem. Res. 44(10), 947–956 (2011).
Medline CASGoogle Scholar
53. Cheng L , Wang C , Feng L , Yang K , Liu Z . Functional nanomaterials for phototherapies of cancer. Chem. Rev. 114(21), 10869–10939 (2014).
Medline CASGoogle Scholar
54. Espinosa A , Di Corato R , Kolosnjaj-Tabi J , Flaud P , Pellegrino T , Wilhelm C . Duality of iron oxide nanoparticles in cancer therapy: amplification of heating efficiency by magnetic hyperthermia and photothermal bimodal treatment. ACS Nano 10(2), 2436–2446 (2016).
Medline CASGoogle Scholar
55. Sheng ZH , Zheng MB , Cai LT . Translational nanomedicine for imaging-guided photothermal therapy. Nanomed. Nanotechnol. 12(2), 568–568 (2016).
Google Scholar
56. Zou LL , Wang H , He B et al. Current approaches of photothermal therapy in treating cancer metastasis with nanotherapeutics. Theranostics 6(6), 762–772 (2016).
Medline CASGoogle Scholar
57. Nam J , Son S , Ochyl LJ , Kuai R , Schwendeman A , Moon JJ . Chemo-photothermal therapy combination elicits anti-tumor immunity against advanced metastatic cancer. Nat. Commun. 9(1), 1074 (2018).
MedlineGoogle Scholar
58. Dong WJ , Li YS , Niu DC et al. Facile synthesis of monodisperse superparamagnetic Fe₃O₄ Core@hybrid@Au shell nanocomposite for bimodal imaging and photothermal therapy. Adv. Mater. 23(45), 5392–5397 (2011).
Medline CASGoogle Scholar
59. Waight JD , Hu Q , Miller A , Liu S , Abrams SI . Tumor-derived G-CSF facilitates neoplastic growth through a granulocytic myeloid-derived suppressor cell-dependent mechanism. PLoS ONE 6(11), e27690 (2011).
Medline CASGoogle Scholar
60. Kachikwu EL , Iwamoto KS , Liao YP et al. Radiation enhances regulatory T cell representation. Int. J. Radiat. Oncol. Biol. Phys. 81(4), 1128–1135 (2011).
MedlineGoogle Scholar
61. Zhang T , Yu HF , Ni C et al. Hypofractionated stereotactic radiation therapy activates the peripheral immune response in operable stage I non-small-cell lung cancer. Sci. Rep. 7, 4866 (2017). • An interesting work revealing the effect of radiation frequency on antitumor efficacy.
Medline CASGoogle Scholar
62. Dewan MZ , Galloway AE , Kawashima N et al. Fractionated but not single-dose radiotherapy induces an immune-mediated abscopal effect when combined with anti-CTLA-4 antibody. Clin. Cancer Res. 15(17), 5379–5388 (2009).
Medline CASGoogle Scholar
63. Schaue D , Ratikan JA , Iwamoto KS , Mcbride WH . Maximizing tumor immunity with fractionated radiation. Int. J. Radiat. Oncol. 83(4), 1306–1310 (2012).
Medline CASGoogle Scholar
64. Lecciso M , Ocadlikova D , Sangaletti S et al. ATP release from chemotherapy-treated dying leukemia cells elicits an immune suppressive effect by increasing regulatory T cells and tolerogenic dendritic cells. Front. Immunol. 8, 1918 (2017).
MedlineGoogle Scholar
65. Li QS , Virtuoso LP , Anderson CD , Egilmez NK . Regulatory rebound in IL-12-treated tumors is driven by uncommitted peripheral regulatory T cells. J. Immunol. 195(3), 1293–1300 (2015).
Medline CASGoogle Scholar
66. Bos PD , Plitas G , Rudra D , Lee SY , Rudensky AY . Transient regulatory T cell ablation deters oncogene-driven breast cancer and enhances radiotherapy. J. Exp. Med. 210(11), 2435–2446 (2013).
MedlineGoogle Scholar
67. Galluzzi L , Buque A , Kepp O , Zitvogel L , Kroemer G . Immunogenic cell death in cancer and infectious disease. Nat. Rev. Immunol. 17(2), 97–111 (2017).
Medline CASGoogle Scholar
68. Medler TR , Cotechini T , Coussens LM . Immune response to cancer therapy: mounting an effective antitumor response and mechanisms of resistance. Trends Cancer 1(1), 66–75 (2015).
MedlineGoogle Scholar
69. Klemm F , Joyce JA . Microenvironmental regulation of therapeutic response in cancer. Trends Cell Biol. 25(4), 198–213 (2015).
MedlineGoogle Scholar
70. Barker HE , Paget JTE , Khan AA , Harrington KJ . The tumour microenvironment after radiotherapy: mechanisms of resistance and recurrence. Nat. Rev. Cancer 15(7), 409–425 (2015).
Medline CASGoogle Scholar
71. Williams CB , Yeh ES , Soloff AC . Tumor-associated macrophages: unwitting accomplices in breast cancer malignancy. NPJ Breast Cancer 2, 15025 (2016).
MedlineGoogle Scholar
72. Martens A , Wistuba-Hamprecht K , Foppen MG et al. Baseline peripheral blood biomarkers associated with clinical outcome of advanced melanoma patients treated with ipilimumab. Clin. Cancer Res. 22(12), 2908–2918 (2016).
Medline CASGoogle Scholar

Vol. 15, No. 1

Supplemental Materials

Metrics

Downloaded 351 times

History

Received 9 May 2019

Accepted 14 October 2019

Published online 23 December 2019

Published in print January 2020

Information

Keywords

Supplementary data

To view the supplementary data that accompany this paper please visit the journal website at: www.futuremedicine.com/doi/suppl/10.2217/nnm-2019-0190

Acknowledgments

The authors thank the technical support from the Center for Molecular Imaging and Biomedical Research Core Facilities at the University of Michigan.

Financial & competing interests disclosure

This work in part was supported by the University of Michigan Comprehensive Cancer Center grant (NIH 2P30CA046592-24 to D Sun), Upjohn Research Award (G021782 to H Chen), UMOR Award (29208 to H Chen), and MCubed Award (U064006 to H Chen). This work was also partially supported by the Gillson Longenbaugh Foundation (to Q Li and A Chang). The authors have no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed.

No writing assistance was utilized in the production of this manuscript.

Ethical conduct of research

The authors state that they have obtained appropriate institutional review board approval or have followed the principles outlined in the Declaration of Helsinki for all human or animal experimental investigations.

PDF download