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01-04-2018 | Original Article

N-acetyl cysteine protects anti-melanoma cytotoxic T cells from exhaustion induced by rapid expansion via the downmodulation of Foxo1 in an Akt-dependent manner

Authors: Matthew J. Scheffel, Gina Scurti, Megan M. Wyatt, Elizabeth Garrett-Mayer, Chrystal M. Paulos, Michael I. Nishimura, Christina Voelkel-Johnson

Published in: Cancer Immunology, Immunotherapy | Issue 4/2018

Abstract

Therapeutic outcomes for adoptive cell transfer (ACT) therapy are constrained by the quality of the infused T cells. The rapid expansion necessary to obtain large numbers of cells results in a more terminally differentiated phenotype with decreased durability and functionality. N-acetyl cysteine (NAC) protects against activation-induced cell death (AICD) and improves anti-tumor efficacy of Pmel-1 T cells in vivo. Here, we show that these benefits of NAC can be extended to engineered T cells and significantly increases T-cell survival within the tumor microenvironment. The addition of NAC to the expansion protocol of human TIL13838I TCR-transduced T cells that are under evaluation in a Phase I clinical trial, demonstrated that findings in murine cells extend to human cells. Expansion of TIL13838I TCR-transduced T cells in NAC also increased their ability to kill target cells in vitro. Interestingly, NAC did not affect memory subsets, but diminished up-regulation of senescence (CD57) and exhaustion (PD-1) markers and significantly decreased expression of the transcription factors EOMES and Foxo1. Pharmacological inhibition of the PI3K/Akt pathway ablates the decrease in Foxo1 induced by NAC treatment of activated T cells. This suggests a model in which NAC through PI3K/Akt activation suppresses Foxo1 expression, thereby impacting its transcriptional targets EOMES, PD-1, and granzyme B. Taken together, our results indicate that NAC exerts pleiotropic effects that impact the quality of TCR-transduced T cells and suggest that the addition of NAC to current clinical protocols should be considered.

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DeSantis C, Lin C, Mariotto A et al (2014) Cancer treatment and survivorship statistics, 2014. CA Cancer J Clin 64:252–271CrossRefPubMed

Rosenberg S (2012) Raising the bar: the curative potential of human cancer immunotherapy. Sci Transl Med 4:127ps8CrossRefPubMed

Johnson LA, June CH (2016) Driving gene-engineered T cell immunotherapy of cancer. Cell Res 38–58

Tran KQ, Zhou J, Durflinger KH et al (2008) Minimally cultured tumor-infiltrating lymphocytes display optimal characteristics for adoptive cell therapy. J Immunother 31:742–751CrossRefPubMedPubMedCentral

Powell DJ, Dudley ME, Robbins PF, Rosenberg SA (2005) Transition of late-stage effector T cells to CD27⁺ CD28⁺ tumor-reactive effector memory T cells in humans after adoptive cell transfer therapy. Blood 105:241–250CrossRefPubMed

Li Y, Liu S, Hernandez J et al (2010) MART-1-specific melanoma tumor-infiltrating lymphocytes maintaining CD28 expression have improved survival and expansion capability following antigenic restimulation in vitro. J Immunol 184:452–465CrossRefPubMed

Ahmadzadeh M, Johnson LA, Heemskerk B et al (2009) Tumor antigen-specific CD8 T cells infiltrating the tumor express high levels of PD-1 and are functionally impaired. Blood 114:1537–1544CrossRefPubMedPubMedCentral

Hernandez-Chacon JA, Li Y, Wu RC et al (2011) Costimulation through the CD137/4-1BB pathway protects human melanoma tumor-infiltrating lymphocytes from activation-induced cell death and enhances antitumor effector function. J Immunother 34:236–250CrossRefPubMedPubMedCentral

Scheffel MJ, Scurti G, Simms P et al (2016) Efficacy of adoptive T-cell therapy is improved by treatment with the antioxidant N-acetyl cysteine, which limits activation-induced T-cell death. Cancer Res 76:6006–6016CrossRefPubMedPubMedCentral

10.

Roszkowski JJ, Lyons GE, Kast WM et al (2005) Simultaneous generation of CD8⁺ and CD4⁺ melanoma-reactive T cells by retroviral-mediated transfer of a single T-cell receptor. Cancer Res 65:1570–1576CrossRefPubMed

11.

Norell H, Zhang Y, McCracken J et al (2010) CD34-based enrichment of genetically engineered human T cells for clinical use results in dramatically enhanced tumor targeting. Cancer Immunol Immunother 59:851–862CrossRefPubMedPubMedCentral

12.

Overwijk WW, Theoret MR, Finkelstein SE et al (2003) Tumor regression and autoimmunity after reversal of a functionally tolerant state of self-reactive CD8⁺ T cells. J Exp Med 198:569–580CrossRefPubMedPubMedCentral

13.

Kerkar SP, Sanchez-Perez L, Borman Z et al (2011) Genetic engineering of murine CD8⁺ and CD4⁺ T cells for preclinical adoptive immunotherapy studies. J Immunother 34:343–352CrossRefPubMedPubMedCentral

14.

Karlsson H, Nava S, Remberger M et al (2011) N-acetyl-l-cysteine increases acute graft-versus-host disease and promotes T-cell-mediated immunity in vitro. Eur J Immunol 41:1143–1153CrossRefPubMed

15.

Moore T, Wagner CR, Scurti GM et al (2017) Clinical and immunologic evaluation of three metastatic melanoma patients treated with autologous melanoma-reactive TCR-transduced T cells. Cancer Immunol Immunother. https://doi.org/10.1007/s00262-017-2073-0

16.

Kesarwani P, Al-Khami AA, Scurti G et al (2014) Promoting thiol expression increases the durability of antitumor T-cell functions. Cancer Res 74:6036–6047CrossRefPubMedPubMedCentral

17.

Klebanoff CA, Gattinoni L, Torabi-Parizi P et al (2005) Central memory self/tumor-reactive CD8⁺ T cells confer superior antitumor immunity compared with effector memory T cells. Proc Natl Acad Sci USA 102:9571–9576CrossRefPubMedPubMedCentral

18.

McLane LM, Banerjee PP, Cosma GL et al (2013) Differential localization of T-bet and Eomes in CD8 T-cell memory populations. J Immunol 190:3207–3215CrossRefPubMedPubMedCentral

19.

Pauken KE, Wherry EJ (2015) Overcoming T cell exhaustion in infection and cancer. Trends Immunol 36:265–276CrossRefPubMedPubMedCentral

20.

Crawford A, Wherry EJ (2009) The diversity of costimulatory and inhibitory receptor pathways and the regulation of antiviral T cell responses. Curr Opin Immunol 21:179–186CrossRefPubMedPubMedCentral

21.

Barber DL, Wherry EJ, Masopust D et al (2006) Restoring function in exhausted CD8 T cells during chronic viral infection. Nature 439:682–687CrossRefPubMed

22.

Staron MM, Gray SM, Marshall HD et al (2014) The transcription factor FoxO1 sustains expression of the inhibitory receptor PD-1 and survival of antiviral CD8⁺ T cells during chronic infection. Immunity 41:802–814CrossRefPubMedPubMedCentral

23.

Rao RR, Li Q, Bupp MRG, Shrikant PA (2012) Transcription factor Foxo1 represses T-bet-mediated effector functions and promotes memory CD8⁺ T cell differentiation. Immunity 36:374–387CrossRefPubMedPubMedCentral

24.

Aoki M, Jiang H, Vogt PK (2004) Proteasomal degradation of the FoxO1 transcriptional regulator in cells transformed by the P3k and Akt oncoproteins. Proc Natl Acad Sci USA 101:13613–13617CrossRefPubMedPubMedCentral

25.

Zaks TZ, Chappell DB, Rosenberg SA, Restifo NP (1999) Fas-mediated suicide of tumor-reactive T cells following activation by specific tumor: selective rescue by caspase inhibition. J Immunol 162:3273–3279PubMedPubMedCentral

26.

Lu B, Finn OJ (2008) T-cell death and cancer immune tolerance. Cell Death Differ 15:70–79CrossRefPubMed

27.

Saff RR, Spanjaard ES, Hohlbaum AM, Marshak-Rothstein A (2004) Activation-induced cell death limits effector function of CD4 tumor-specific T cells. J Immunol 172:6598–6606CrossRefPubMed

28.

Robbins PF, Dudley ME, Wunderlich J et al (2004) Cutting edge: persistence of transferred lymphocyte clonotypes correlates with cancer regression in patients receiving cell transfer therapy. J Immunol 173:7125–7130CrossRefPubMedPubMedCentral

29.

Klebanoff CA, Gattinoni L, Restifo NP (2012) Sorting through subsets: which T-cell populations mediate highly effective adoptive immunotherapy? J Immunother 35:651–660CrossRefPubMedPubMedCentral

30.

Gattinoni L, Klebanoff CA, Palmer DC et al (2005) Acquisition of full effector function in vitro paradoxically impairs the in vivo antitumor efficacy of adoptively transferred CD8⁺ T cells. J Clin Invest 115:1616–1626CrossRefPubMedPubMedCentral

31.

Brenchley JM, Karandikar NJ, Betts MR et al (2003) Expression of CD57 defines replicative senescence and antigen-induced apoptotic death of CD8. Blood 101:2711–2720CrossRefPubMed

32.

Dudley ME, Yang JC, Sherry R et al (2008) Adoptive cell therapy for patients with metastatic melanoma: evaluation of intensive myeloablative chemoradiation preparative regimens. J Clin Oncol 26:5233–5239CrossRefPubMedPubMedCentral

33.

Shen X, Zhou J, Hathcock KS et al (2007) Persistence of tumor infiltrating lymphocytes in adoptive immunotherapy correlates with telomere length. J Immunother 30:123–129CrossRefPubMedPubMedCentral

34.

Zhou J, Shen X, Huang J et al (2005) Telomere length of transferred lymphocytes correlates with in vivo persistence and tumor regression in melanoma patients receiving cell transfer therapy. J Immunol 175:7046–7052CrossRefPubMedPubMedCentral

35.

Hsin IL, Sheu GT, Chen HH et al (2010) N-acetyl cysteine mitigates curcumin-mediated telomerase inhibition through rescuing of Sp1 reduction in A549 cells. Mutat Res 688:72–77CrossRefPubMed

36.

Liu J, Liu M, Ye X et al (2012) Delay in oocyte aging in mice by the antioxidant N-acetyl-l-cysteine (NAC). Hum Reprod 27:1411–1420CrossRefPubMed

37.

Haendeler J, Hoffmann J, Diehl JF et al (2004) Antioxidants inhibit nuclear export of telomerase reverse transcriptase and delay replicative senescence of endothelial cells. Circ Res 94:768–775CrossRefPubMed

38.

Hao LY, Strong MA, Greider CW (2004) Phosphorylation of H2AX at short telomeres in T cells and fibroblasts. J Biol Chem 279:45148–45154CrossRefPubMed

39.

Kerdiles YM, Beisner DR, Tinoco R et al (2009) Foxo1 links homing and survival of naive T cells by regulating l-selectin, CCR7 and interleukin 7 receptor. Nat Immunol 10:176–184CrossRefPubMedPubMedCentral

40.

Matsuzaki H, Daitoku H, Hatta M et al (2003) Insulin-induced phosphorylation of FKHR (Foxo1) targets to proteasomal degradation. Proc Natl Acad Sci USA 100:11285–11290CrossRefPubMedPubMedCentral

41.

Noh YH, Chob HS, Kim DH et al (2012) N-acetylcysteine enhances neuronal differentiation of P19 embryonic stem cells via Akt and N-cadherin activation. Mol Biol (Mosk) 46:741–746CrossRef

42.

Wang T, Mao X, Li H et al (2013) N-Acetylcysteine and allopurinol up-regulated the Jak/STAT3 and PI3K/Akt pathways via adiponectin and attenuated myocardial postischemic injury in diabetes. Free Radic Biol Med 63:291–303CrossRefPubMed

43.

Wang C, Xia Y, Zheng Y et al (2015) Protective effects of N-acetylcysteine in concanavalin a-induced hepatitis in mice. Mediators Inflamm. https://doi.org/10.1155/2015/189785 PubMedPubMedCentral

44.

Jin HM, Zhou DC, Gu HF et al (2013) Antioxidant N-acetylcysteine protects pancreatic β-cells against aldosterone-induced oxidative stress and apoptosis in female db/db mice and insulin-producing MIN6 cells. Endocrinology 154:4068–4077CrossRefPubMed

45.

Leslie NR (2006) The redox regulation of PI 3-kinase-dependent signaling. Antioxid Redox Signal 8:1765–1774CrossRefPubMed

46.

Leslie NR, Bennett D, Lindsay YE et al (2003) Redox regulation of PI 3-kinase signalling via inactivation of PTEN. EMBO J 22:5501–5510CrossRefPubMedPubMedCentral

47.

Ahmad F, Nidadavolu P, Durgadoss L et al (2014) Critical cysteines in Akt1 regulate its activity and proteasomal degradation: implications for neurodegenerative diseases. Free Radic Biol Med 74:118 – 28CrossRefPubMed

48.

Kaech SM, Cui W (2012) Transcriptional control of effector and memory CD8⁺ T cell differentiation. Nat Rev Immunol 12:749–761CrossRefPubMedPubMedCentral

Title: N-acetyl cysteine protects anti-melanoma cytotoxic T cells from exhaustion induced by rapid expansion via the downmodulation of Foxo1 in an Akt-dependent manner
Authors: Matthew J. Scheffel
Gina Scurti
Megan M. Wyatt
Elizabeth Garrett-Mayer
Chrystal M. Paulos
Michael I. Nishimura
Christina Voelkel-Johnson
Publication date: 01-04-2018
Publisher: Springer Berlin Heidelberg
Published in: Cancer Immunology, Immunotherapy / Issue 4/2018
Print ISSN: 0340-7004
Electronic ISSN: 1432-0851
DOI: https://doi.org/10.1007/s00262-018-2120-5

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