Immunological mechanisms of the antitumor effects of supplemental oxygenation
Paving the way for intratumoral T cells
Tumors often express unusual antigens and are surrounded by immune cells. Unfortunately, this immune surveillance is imperfect and does not always prevent the tumors from growing. In addition, tumors are often hypoxic, because their rapid growth outstrips that of their blood and oxygen supply. Now, Hatfield et al. have linked these two phenomena by demonstrating that T cells avoid going into the hypoxic areas of tumors. The authors have also shown a way to overcome this problem in mice with lung tumors by having the animals breathe supplementary oxygen. Having a higher concentration of oxygen throughout the body improved the oxygenation inside the tumors, allowing immune cells to enter the tumors and attack them, extending the animals’ survival.
Abstract
Antitumor T cells either avoid or are inhibited in hypoxic and extracellular adenosine-rich tumor microenvironments (TMEs) by A2A adenosine receptors. This may limit further advances in cancer immunotherapy. There is a need for readily available and safe treatments that weaken the hypoxia–A2-adenosinergic immunosuppression in the TME. Recently, we reported that respiratory hyperoxia decreases intratumoral hypoxia and concentrations of extracellular adenosine. We show that it also reverses the hypoxia-adenosinergic immunosuppression in the TME. This, in turn, stimulates (i) enhanced intratumoral infiltration and reduced inhibition of endogenously developed or adoptively transfered tumor-reactive CD8 T cells, (ii) increased proinflammatory cytokines and decreased immunosuppressive molecules, such as transforming growth factor–β (TGF-β), (iii) weakened immunosuppression by regulatory T cells, and (iv) improved lung tumor regression and long-term survival in mice. Respiratory hyperoxia also promoted the regression of spontaneous metastasis from orthotopically grown breast tumors. These effects are entirely T cell– and natural killer cell–dependent, thereby justifying the testing of supplemental oxygen as an immunological coadjuvant to combine with existing immunotherapies for cancer.
Get full access to this article
View all available purchase options and get full access to this article.
Supplementary Material
Summary
Materials and Methods
Fig. S1. The tumor-regressing effects of respiratory hyperoxia are lost in cγ/Rag-2−/− mice.
Fig. S2. ROS scavenger does not prevent the antitumor effects of respiratory hyperoxia.
Fig. S3. Respiratory hyperoxia reverses hypoxia-adenosinergic inhibition of NK cells.
Fig. S4. Respiratory hyperoxia does not further improve the activity of tumor-reactive A2AR−/− T cells.
Fig. S5. CD8 and CD4 T cells avoid hypoxic TME.
Fig. S6. Tregs with higher expression of CTLA-4 are more hypoxic.
Fig. S7. CD8 T cells from TDLN are enriched after culture activation for adoptive transfer.
Fig. S8. Breathing 60% oxygen increased IFN-γ production by CD8 T cells in the lung TME.
Table S1. Immunostimulating cytokines/chemokines increased by respiratory hyperoxia.
Table S2. Full list of primer sets in RT-PCR arrays.
Resources
File (7-277ra30_sm.pdf)
- Download
- 4.08 MB
REFERENCES AND NOTES
1
Ohta A., Sitkovsky M., Role of G-protein-coupled adenosine receptors in downregulation of inflammation and protection from tissue damage. Nature 414, 916–920 (2001).
2
Ohta A., Gorelik E., Prasad S. J., Ronchese F., Lukashev D., Wong M. K. K., Huang X., Caldwell S., Liu K., Smith P., Chen J.-F., Jackson E. K., Apasov S., Abrams S., Sitkovsky M., A2A adenosine receptor protects tumors from antitumor T cells. Proc. Natl. Acad. Sci. U.S.A. 103, 13132–13137 (2006).
3
Eltzschig H. K., Sitkovsky M. V., Robson S. C., Purinergic signaling during inflammation. N. Engl. J. Med. 367, 2322–2333 (2012).
4
Raskovalova T., Huang X., Sitkovsky M., Zacharia L. C., Jackson E. K., Gorelik E., Gs protein-coupled adenosine receptor signaling and lytic function of activated NK cells. J. Immunol. 175, 4383–4391 (2005).
5
Sitkovsky M. V., T regulatory cells: Hypoxia-adenosinergic suppression and re-direction of the immune response. Trends Immunol. 30, 102–108 (2009).
6
Sitkovsky M. V., Hatfield S., Abbott R., Belikoff B., Lukashev D., Ohta A., Hostile, hypoxia–A2-adenosinergic tumor biology as the next barrier to overcome for tumor immunologists. Cancer Immunol. Res. 2, 598–605 (2014).
7
Rosenberg S. A., Restifo N. P., Yang J. C., Morgan R. A., Dudley M. E., Adoptive cell transfer: A clinical path to effective cancer immunotherapy. Nat. Rev. Cancer 8, 299–308 (2008).
8
Sharma P., Wagner K., Wolchok J. D., Allison J. P., Novel cancer immunotherapy agents with survival benefit: Recent successes and next steps. Nat. Rev. Cancer 11, 805–812 (2011).
9
Naidoo J., Page D. B., Wolchok J. D., Immune modulation for cancer therapy. Br. J. Cancer 111, 2214–2219 (2014).
10
Mellman I., Coukos G., Dranoff G., Cancer immunotherapy comes of age. Nature 480, 480–489 (2011).
11
Hodi F. S., O’Day S. J., McDermott D. F., Weber R. W., Sosman J. A., Haanen J. B., Gonzalez R., Robert C., Schadendorf D., Hassel J. C., Akerley W., van den Eertwegh A. J., Lutzky J., Lorigan P., Vaubel J. M., Linette G. P., Hogg D., Ottensmeier C. H., Lebbé C., Peschel C., Quirt I., Clark J. I., Wolchok J. D., Weber J. S., Tian J., Yellin M. J., Nichol G. M., Hoos A., Urba W. J., Improved survival with ipilimumab in patients with metastatic melanoma. N. Engl. J. Med. 363, 711–723 (2010).
12
Pardoll D. M., The blockade of immune checkpoints in cancer immunotherapy. Nat. Rev. Cancer 12, 252–264 (2012).
13
Young A., Mittal D., Stagg J., Smyth M. J., Targeting cancer-derived adenosine: New therapeutic approaches. Cancer Discov. 4, 879–888 (2014).
14
Koebel C. M., Vermi W., Swann J. B., Zerafa N., Rodig S. J., Old L. J., Smyth M. J., Schreiber R. D., Adaptive immunity maintains occult cancer in an equilibrium state. Nature 450, 903–907 (2007).
15
Waickman A. T., Alme A., Senaldi L., Zarek P. E., Horton M., Powell J. D., Enhancement of tumor immunotherapy by deletion of the A2A adenosine receptor. Cancer Immunol. Immunother. 61, 917–926 (2012).
16
Künzli B. M., Bernlochner M. I., Rath S., Käser S., Csizmadia E., Enjyoji K., Cowan P., d’Apice A., Dwyer K., Rosenberg R., Perren A., Friess H., Maurer C. A., Robson S. C., Impact of CD39 and purinergic signalling on the growth and metastasis of colorectal cancer. Purinergic Signal. 7, 231–241 (2011).
17
Stagg J., Divisekera U., McLaughlin N., Sharkey J., Pommey S., Denoyer D., Dwyer K. M., and, Smyth M. J., Anti-CD73 antibody therapy inhibits breast tumor growth and metastasis. Proc. Natl. Acad. Sci. U.S.A. 107, 1547–1552 (2010).
18
Stagg J., Divisekera U., Duret H., Sparwasser T., Teng M. W. L., Darcy P. K., Smyth M. J., CD73-deficient mice have increased antitumor immunity and are resistant to experimental metastasis. Cancer Res. 71, 2892–2900 (2011).
19
Jin D., Fan J., Wang L., Thompson L. F., Liu A., Daniel B. J., Shin T., Curiel T. J., Zhang B., CD73 on tumor cells impairs antitumor T-cell responses: A novel mechanism of tumor-induced immune suppression. Cancer Res. 70, 2245–2255 (2010).
20
Beavis P. A., Divisekera U., Paget C., Chow M. T., John L. B., Devaud C., Dwyer K., Stagge J., Smyth M. J., Darcy P. K., Blockade of A2A receptors potently suppresses the metastasis of CD73+ tumors. Proc. Natl. Acad. Sci. U.S.A. 110, 14711–14716 (2013).
21
Loi S., Pommey S., Haibe-Kains B., Beavis P. A., Darcy P. K., Smyth M. J., Stagg J., CD73 promotes anthracycline resistance and poor prognosis in triple negative breast cancer. Proc. Natl. Acad. Sci. U.S.A. 110, 11091–11096 (2013).
22
Kallet R. H., Matthay M. A., Hyperoxic acute lung injury. Respir. Care 58, 123–141 (2013).
23
Thiel M., Chouker A., Ohta A., Jackson E., Caldwell C., Smith P., Lukashev D., Bittmann I., Sitkovsky M. V., Oxygenation inhibits the physiological tissue-protecting mechanism and thereby exacerbates acute inflammatory lung injury. PLOS Biol. 3, e174 (2005).
24
Vaupel P., Tumor oxygenation: An appraisal of past and present concepts and a look into the future: Arisztid G. B. Kovách Lecture. Adv. Exp. Med. Biol. 789, 229–236 (2013).
25
Hatfield S. M., Kjaergaard J., Lukashev D., Belikoff B., Schreiber T. H., Sethumadhavan S., Abbott R., Philbrook P., Thayer M., Shujia D., Rodig S., Kutok J. L., Ren J., Ohta A., Podack E. R., Karger B., Jackson E. K., Sitkovsky M., Systemic oxygenation weakens the hypoxia and hypoxia inducible factor 1α-dependent and extracellular adenosine-mediated tumor protection. J. Mol. Med. 92, 1283–1292 (2014).
26
Dewhirst M. W., Cao Y., Moeller B., Cycling hypoxia and free radicals regulate angiogenesis and radiotherapy response. Nat. Rev. Cancer 8, 425–437 (2008).
27
Wang J., Yi J., Cancer cell killing via ROS: To increase or decrease, that is the question. Cancer Biol. Ther. 7, 1875–1884 (2008).
28
Min J. H., Codipilly C. N., Nasim S., Miller E. J., Ahmed M. N., Synergistic protection against hyperoxia-induced lung injury by neutrophils blockade and EC-SOD overexpression. Respir. Res. 13, 58 (2012).
29
Kratzer E., Tian Y., Sarich N., Wu T., Meliton A., Leff A., Birukova A. A., Oxidative stress contributes to lung injury and barrier dysfunction via microtubule destabilization. Am. J. Respir. Cell Mol. Biol. 47, 688–697 (2012).
30
Kjaergaard J., Shu S., Tumor infiltration by adoptively transferred T cells is independent of immunologic specificity but requires down-regulation of L-selectin expression. J. Immunol. 163, 751–759 (1999).
31
Kjaergaard J., Peng L., Cohen P. A., Shu S., Therapeutic efficacy of adoptive immunotherapy is predicated on in vivo antigen-specific proliferation of donor T cells. Clin. Immunol. 108, 8–20 (2003).
32
Champsaur M., Lanier L. L., Effect of NKG2D ligand expression on host immune responses. Immunol. Rev. 235, 267–285 (2010).
33
Raulet D. H., Gasser S., Gowen B. G., Deng W., Jung H., Regulation of ligands for the NKG2D activating receptor. Annu. Rev. Immunol. 31, 413–441 (2013).
34
Quezada S. A., Peggs K. S., Simpson T. R., Shen Y., Littman D. R., Allison J. P., Limited tumor infiltration by activated T effector cells restricts the therapeutic activity of regulatory T cell depletion against established melanoma. J. Exp. Med. 205, 2125–2138 (2008).
35
Schreiber T. H., Deyev V. V., Rosenblatt J. D., Podack E. R., Tumor-induced suppression of CTL expansion and subjugation by gp96-Ig vaccination. Cancer Res. 69, 2026–2033 (2009).
36
Sitkovsky M. V., Kjaergaard J., Lukashev D., Ohta A., Hypoxia-adenosinergic immunosuppression: Tumor protection by T regulatory cells and cancerous tissue hypoxia. Clin. Cancer Res. 14, 5947–5952 (2008).
37
Wing K., Onishi Y., Prieto-Martin P., Yamaguchi T., Miyara M., Fehervari Z., Nomura T., Sakaguchi S., CTLA-4 control over Foxp3+ regulatory T cell function. Science 322, 271–275 (2008).
38
Kjaergaard J., Peng L., Cohen P. A., Drazba J. A., Weinberg A. D., Shu S., Augmentation versus inhibition: Effects of conjunctional OX-40 receptor monoclonal antibody and IL-2 treatment on adoptive immunotherapy of advanced tumor. J. Immunol. 167, 6669–6677 (2001).
39
Koshiba M., Kojima H., Huang S., Apasov S., Sitkovsky M. V., Memory of extracellular adenosine A2A purinergic receptor-mediated signaling in murine T cells. J. Biol. Chem. 272, 25881–25889 (1997).
40
Kobayashi S., Zimmermann H., Millhorn D. E., Chronic hypoxia enhances adenosine release in rat PC12 cells by altering adenosine metabolism and membrane transport. J. Neurochem. 74, 621–632 (2000).
41
Decking U. K., Schlieper G., Kroll K., Schrader J., Hypoxia-induced inhibition of adenosinekinase potentiates cardiac adenosine release. Circ. Res. 81, 154–164 (1997).
42
Synnestvedt K., Furuta G. T., Comerford K. M., Louis N., Karhausen J., Eltzschig H. K., Hansen K. R., Thompson L. F., Colgan S. P., Ecto-5'-nucleotidase (CD73) regulation by hypoxia-inducible factor-1 mediates permeability changes in intestinal epithelia. J. Clin. Invest. 110, 993–1002 (2002).
43
Corzo C. A., Condamine T., Lu L., Cotter M. J., Youn J. I., Cheng P., Cho H. I., Celis E., Quiceno D. G., Padhya T., McCaffrey T. V., McCaffrey J. C., Gabrilovich D. I., HIF-1α regulates function and differentiation of myeloid-derived suppressor cells in the tumor microenvironment. J. Exp. Med. 207, 2439–2453 (2010).
44
Colegio O. R., Chu N.-Q., Szabo A. L., Chu T., Marie Rhebergen A., Jairam V., Cyrus N., Brokowski C. E., Eisenbarth S. C., Phillips G. M., Cline G. W., Phillips A. J., Medzhitov R., Functional polarization of tumour-associated macrophages by tumour-derived lactic acid. Nature 513, 559–563 (2014).
45
Chaturvedi P., Gilkes D. M., Takano N., Semenza G. L., Hypoxia-inducible factor-dependent signaling between triple-negative breast cancer cells and mesenchymal stem cells promotes macrophage recruitment. Proc. Natl. Acad. Sci. U.S.A. 111, E2120–E2129 (2014).
46
Ohta A., Sitkovsky M., Caveats in promising therapeutic targeting of the anti-inflammatory A2 adenosine receptors: The notes of caution. Nat. Rev. Drug Discov. 5 (2006).
47
Ooi H., Cadogan E., Sweeney M., Howell K., O’Regan R. G., McLoughlin P., Chronic hypercapnia inhibits hypoxic pulmonary vascular remodeling. Am. J. Physiol. Heart Circ. Physiol. 278, H331–H338 (2000).
48
De Smet H. R., Bersten A. D., Barr H. A., Doyle I. R., Hypercapnic acidosis modulates inflammation, lung mechanics, and edema in the isolated perfused lung. J. Crit. Care 22, 305–313 (2007).
49
Kjaergaard J., Tanaka J., Kim J. A., Rothchild K., Weinberg A., Shu S., Therapeutic efficacy of OX-40 receptor antibody depends on tumor immunogenicity and anatomic site of tumor growth. Cancer Res. 60, 5514–5521 (2000).
(0)eLetters
eLetters is a forum for ongoing peer review. eLetters are not edited, proofread, or indexed, but they are screened. eLetters should provide substantive and scholarly commentary on the article. Embedded figures cannot be submitted, and we discourage the use of figures within eLetters in general. If a figure is essential, please include a link to the figure within the text of the eLetter. Please read our Terms of Service before submitting an eLetter.
Log In to Submit a ResponseNo eLetters have been published for this article yet.
Information & Authors
Information
Published In
Science Translational Medicine
Volume 7 | Issue 277
March 2015
March 2015
Copyright
Copyright © 2015, American Association for the Advancement of Science.
Submission history
Received: 23 October 2014
Accepted: 28 January 2015
Acknowledgments
We thank J. Stagg at the University of Montreal for providing the 4T1 tumor cell line and sharing his expertise. We thank S. Ohman for assistance in all steps leading to the preparation of the manuscript. We also thank R. Marsh, professor of biology at Northeastern University, for assistance with monitoring and controlling gas levels in hyperoxic units. Funding: This work was supported by funding from Northeastern University and NIH grants to M.V.S. (R01 CA 112561, R01 CA 111985, R21 AT 002788, U19 AI 091693, Dana-Farber Cancer Institute, and Harvard Medical School–Northeastern University Joint Program in Cancer Drug Development) as well as by National Cancer Institute grant 5PO1CA109094-03 and National Institute of Allergy and Infectious Diseases 1P01 grant AI096396-01 3 to E.R.P.; HL109002, DK091190, DK068575, DK079307, and CA168628 to E.K. J.; and a Bankhead-Coley Postdoctoral Fellowship to T.H.S. Author contributions: S.M.H., J.K., A.O., and M.V.S. performed and/or analyzed cancer immunology assays. S.R. and J.L.K. performed, enumerated, and interpreted immunohistochemistry assays. J.K. established and supervised lung tumor models. E.R.P. and T.H.S. designed and performed assays with custom-made RNA arrays to scan lung TME. D.L., B.B., R.A., S.S., P.P., K.K., R.C., and M.T. performed or assisted in tumor immunology, immunohistochemistry, and flow cytometry assays. E.K.J. and B.K. analyzed and interpreted changes in the TME. M.V.S. designed the overall approach, directed all stages of research, and wrote the manuscript with S.M.H. Competing interests: U.S. government, NIH holds an issued patent “Methods for using extracellular adenosine inhibitors and adenosine receptor inhibitors to enhance immune response and inflammation,” US 8,716,301, which is related to the work described in this paper, with A.O. and M.V.S. named as inventors. M.V.S. is a founder of RedoxTherapies, a company that is charged with the translation of this approach into the clinic and has licensed this patent. E.R.P. has a provisional patent for “Anti-immune suppression tumor therapy” and is the scientific cofounder, paid consultant, and equity owner of Heat Biologics Inc., and T.H.S. is an employee at Heat Biologics Inc. J.L.K. is an employee and shareholder at Infinity Pharmaceuticals Inc. E.K.J. holds equity in Adenopaint, which is unrelated to the current study. Data and materials availability: Correspondence and requests for materials should be addressed to M.V.S. ([email protected]).
Authors
Metrics & Citations
Metrics
Article Usage
Altmetrics
Citations
Cite as
- Stephen M. Hatfield et al.
Export citation
Select the format you want to export the citation of this publication.
Cited by
- Hypoxia, a Targetable Culprit to Counter Pancreatic Cancer Resistance to Therapy, Cancers, 15, 4, (1235), (2023).https://doi.org/10.3390/cancers15041235
- Purinergic signaling: Diverse effects and therapeutic potential in cancer, Frontiers in Oncology, 13, (2023).https://doi.org/10.3389/fonc.2023.1058371
- Towards a consensus definition of immune exclusion in cancer, Frontiers in Immunology, 14, (2023).https://doi.org/10.3389/fimmu.2023.1084887
- Hypoxia-responsive circular RNA circAAGAB reduces breast cancer malignancy by activating p38 MAPK and sponging miR-378 h, Cancer Cell International, 23, 1, (2023).https://doi.org/10.1186/s12935-023-02891-0
- Characterization of Oxygen Nanobubbles and In Vitro Evaluation of Retinal Cells in Hypoxia, Translational Vision Science & Technology, 12, 2, (16), (2023).https://doi.org/10.1167/tvst.12.2.16
- Hypoxic microenvironment in cancer: molecular mechanisms and therapeutic interventions, Signal Transduction and Targeted Therapy, 8, 1, (2023).https://doi.org/10.1038/s41392-023-01332-8
- Nanobiomaterials to modulate natural killer cell responses for effective cancer immunotherapy, Trends in Biotechnology, 41, 1, (77-92), (2023).https://doi.org/10.1016/j.tibtech.2022.06.011
- Prognostic impact of CD73/adenosine 2A receptor (A2AR) in renal cell carcinoma and immune microenvironmental status with sarcomatoid changes and rhabdoid features, Pathology - Research and Practice, 244, (154423), (2023).https://doi.org/10.1016/j.prp.2023.154423
- Oxygen therapy accelerates apoptosis induced by selenium compounds via regulating Nrf2/MAPK signaling pathway in hepatocellular carcinoma, Pharmacological Research, 187, (106624), (2023).https://doi.org/10.1016/j.phrs.2022.106624
- A macrophage membrane-coated mesoporous silica nanoplatform inhibiting adenosine A2AR via in situ oxygen supply for immunotherapy, Journal of Controlled Release, 353, (535-548), (2023).https://doi.org/10.1016/j.jconrel.2022.12.001
- See more
Loading...
View Options
Check Access
Log in to view the full text
AAAS login provides access to Science for AAAS Members, and access to other journals in the Science family to users who have purchased individual subscriptions.
- Become a AAAS Member
- Activate your AAAS ID
- Purchase Access to Other Journals in the Science Family
- Account Help
Log in via OpenAthens.
Log in via Shibboleth.
More options
Register for free to read this article
As a service to the community, this article is available for free. Login or register for free to read this article.
Oxygen and Human Viruses
Dear Editor , We have found very interesting this article of Stephen Hatfield suggesting the anti-tumoral effect of hyperoxia.We suggest that oxygen supplementation might be also useful as an adjuvant therapy for the treatment of chronic viral infections. Effectively ,hypoxia (1% oxygen) up regulates in vitro the expression of some human DNA and RNA viruses(review in reference1); also oxygenotherapy (hyperoxia) might be useful to down load gene expression for these viruses.
Very interestingly Victoria Polonis described in 1991 that hypoxia up regulates in vitro at 15 fold level the replication of HIV-1 expression(2). AIDS is a chronic viral infection and despite intensive research there is no vaccine; consequently the control of HIV-1 viral load is very important but has some limit despite the large implementation of tritherapy.Also we suggest that the use of supplemental oxygenation in addition to tritherapy will merit further consideration in some AIDS patients as described in this paper for cancer treatment.
References (1)Oxygen Tension Level and Human Viral Infections. Frederic Morinet,Luana Casetti,Jean-Hugues Francois, Claude Capron,Sylvie Pillet. Virology 2013;444:31-36 (invited review). (2)Anoxia Induces Human Immunodeficiency Virus Expression in Infected T Cell Lines.Victoria R. Polonis,Garth R.Anderson,Maryanne T.Vahey,Paul J.Morrow,Daniel Stoler,Robert R.Redfield.J. Biol.Chem. 1991;266:11421- 11424.
Frederic Morinet Universite Paris Diderot Paris7 Mathilde Parent Universite de Versailles Saint Quentin en Yvelines Sylvie Pillet Universite Jean Monnet Saint Etienne Claude Capron Universite de Versailles Saint Quentin en Yvelines France