Combination of IL-2, rapamycin, DNA methyltransferase and histone deacetylase inhibitors for the expansion of human regulatory T cells

Makoto Miyara; Driss Chader; Aude Burlion; Jérémie Goldstein; Delphine Sterlin; Françoise Norol; Hélène Trebeden-Nègre; Laetitia Claër; Shimon Sakaguchi; Gilles Marodon; Zahir Amoura; Guy Gorochov

doi:10.18632/oncotarget.10914

Combination of IL-2, rapamycin, DNA methyltransferase and histone deacetylase inhibitors for the expansion of human regulatory T cells

Oncotarget. 2016 Jul 28;8(62):104733-104744. doi: 10.18632/oncotarget.10914. eCollection 2017 Dec 1.

Authors

Makoto Miyara^{1

2}, Driss Chader^{1

2}, Aude Burlion², Jérémie Goldstein², Delphine Sterlin^{1

2}, Françoise Norol³, Hélène Trebeden-Nègre³, Laetitia Claër^{1

2}, Shimon Sakaguchi^#⁴, Gilles Marodon^#², Zahir Amoura^#^{1

2

5

6}, Guy Gorochov^{1

2

6}

Affiliations

¹ Department of immunology, AP-HP Pitié Salpêtrière, Paris, France.
² Sorbonne Universités, UPMC Univ Paris 06, INSERM, CNRS, Centre d'Immunologie et des Maladies Infectieuses (CIMI-Paris), Paris, France.
³ Cell Therapy, AP-HP Pitié Salpêtrière, Paris, France.
⁴ Experimental Immunology, Immunology Frontier Research Center, Osaka University, Osaka, Japan.
⁵ Internal Medicine, French Reference Center for Systemic Lupus Erythematosus and Antiphospholipid Syndrome, AP-HP Pitié Salpêtrière, Paris, France.
⁶ UPMC Paris Sorbonne, Paris, France.

^# Contributed equally.

Abstract

FOXP3⁺ regulatory T cell (Treg) based cellular therapies represent promising therapeutic options in autoimmunity, allergy, transplantation and prevention of Graft Versus Host (GVH) Disease. Among human FOXP3-expressing CD4⁺T cells, only the CD45RA⁺ naïve Treg (nTreg) subset is suitable for in vitro expansion. However, FoxP3 expression decays in cells using currently described culture protocols. Rapamycin alone was not able to prevent FOXP3 loss in nTregs cells, as only a half of them maintained FOXP3 expression after 14 days of culture. In contrast we report a novel combined drug regimen that can drastically stabilize FOXP3 expression in cultured Tregs. IL-2, rapamycin, histone deacetylase and DNA methyltransferase inhibitors act in synergy to allow expansion of human regulatory T cells with sustained high expression of FOXP3 and CD15s with potent suppressive capacities in vitro and control of murine xeno-GVH reactions. Of note, an additional subsequent infusion of expanded nTreg cells did not improve survival of mice. Combination of IL-2, rapamycin, histone deacetylase and DNA methyltransferase inhibitors is optimal for the expansion in vitro of pure effective nTreg maintaining high levels of FOXP3 for therapeutic purposes.

Keywords: FOXP3; GVH; autoimmunity; cell therapy; human regulatory T cells.