Immunomodulatory Effects of Tyrosine Kinase Inhibitors (TKIs) in Vitro and in Vivo Study

Elena MARINELLI BUSILACCHI; Andrea COSTANTINI; Nadia VIOLA; Benedetta COSTANTINI; Jacopo OLIVIERI; Luca BUTINI; Giorgia MANCINI; Ilaria SCORTECHINI; Martina CHIARUCCI; Monica POIANI; Antonella POLONI; Pietro LEONI; Attilio OLIVIERI

doi:10.1016/j.bbmt.2017.10.039

Immunomodulatory Effects of Tyrosine Kinase Inhibitors (TKIs) in Vitro and in Vivo Study

Elena MARINELLI BUSILACCHI, Andrea COSTANTINI, Nadia VIOLA, Benedetta COSTANTINI, Jacopo OLIVIERI, Luca BUTINI, Giorgia MANCINI, Ilaria SCORTECHINI, Martina CHIARUCCI, Monica POIANI, Antonella POLONI, Pietro LEONI, Attilio OLIVIERI

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Abstract

Pathogenesis of chronic Graft Versus Host Disease (cGVHD) is incompletely defined, involving donor-derived CD4 and CD8-positive T lymphocytes, as well as B cells. Standard treatment is lacking for steroid dependent/refractory cases; potential usefulness of tyrosine kinase inhibitors (TKIs) has been suggested, based on their potent anti-fibrotic effect. However, TKI seems to have pleiotropic activity. We sought to evaluate the in vitro and in vivo impact of different TKIs on lymphocyte phenotype and function. Peripheral blood mononuclear cells (PBMC) from healthy donors were cultured in presence of increasing concentrations of Nilotinib, Imatinib, Dasatinib and Ponatinib; in parallel 44 PBMC samples from 15 patients with steroid-dependent/refractory cGVHD treated with Nilotinib in the setting of a phase 1-2 trial were analyzed at baseline, after 90 and after 180 days of therapy. Flow cytometry was performed after labelling lymphocytes with a panel of monoclonal antibodies (CD3, CD4, CD16, CD56, CD25, CD19, CD45RA, FoxP3, CD127 and 7AAD). Cytokine production was assessed in supernatants of purified CD3+ T cells and in plasma samples from Nilotinib-treated patients. Main T lymphocytes subpopulations were not significantly affected by therapeutic concentrations of TKIs in vitro, whereas pro-inflammatory cytokine (in particular IL2, IFNγ, TNFα and IL10) and IL17 production showed a sharp decline. Frequency of T regulatory, B and NK cells decreased progressively in presence of therapeutic concentrations of all the TKIs tested in vitro, except for Nilotinib that showed little effect on these subsets; of note, naïve T regulatory cell subset accumulated following exposure to TKIs. Results obtained in vivo on Nilotinib-treated patients were largely comparable, both on lymphocyte subset kinetics and on cytokine production by CD3-positive cells. This study underlines the anti-inflammatory and immunomodulatory effects of TKIs, and support their potential usefulness as treatment for patients with steroid dependent/refractory cGVHD. In addition, both in vitro and in vivo data point out that, compared to other TKIs, Nilotinib could better preserve the integrity of some important regulatory subsets, such as T regulatory and NK cells.

Original language	English
Journal	BIOLOGY OF BLOOD AND MARROW TRANSPLANTATION
Early online date	8 Nov 2017
DOIs	https://doi.org/10.1016/j.bbmt.2017.10.039
Publication status	E-pub ahead of print - 8 Nov 2017

Keywords

Tyrosine kinase inhibitors (TKIs)
Chronic Graft Versus Host Disease (cGVHD)
Lymphocyte subpopulations
T regulatory cells
Cytokine production

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.1016/j.bbmt.2017.10.039

Immunomodulatory Effects of Tyrosine_BUSILACCHI_PublishedonlineNovember2017_GREEN AAM (CC BY-NC-ND)Accepted author manuscript, 1.39 MBLicence: CC BY-NC-ND

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MARINELLI BUSILACCHI, E., COSTANTINI, A., VIOLA, N., COSTANTINI, B., OLIVIERI, J., BUTINI, L., MANCINI, G., SCORTECHINI, I., CHIARUCCI, M., POIANI, M., POLONI, A., LEONI, P., & OLIVIERI, A. (2017). Immunomodulatory Effects of Tyrosine Kinase Inhibitors (TKIs) in Vitro and in Vivo Study. BIOLOGY OF BLOOD AND MARROW TRANSPLANTATION. Advance online publication. https://doi.org/10.1016/j.bbmt.2017.10.039

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title = "Immunomodulatory Effects of Tyrosine Kinase Inhibitors (TKIs) in Vitro and in Vivo Study",

abstract = "Pathogenesis of chronic Graft Versus Host Disease (cGVHD) is incompletely defined, involving donor-derived CD4 and CD8-positive T lymphocytes, as well as B cells. Standard treatment is lacking for steroid dependent/refractory cases; potential usefulness of tyrosine kinase inhibitors (TKIs) has been suggested, based on their potent anti-fibrotic effect. However, TKI seems to have pleiotropic activity. We sought to evaluate the in vitro and in vivo impact of different TKIs on lymphocyte phenotype and function. Peripheral blood mononuclear cells (PBMC) from healthy donors were cultured in presence of increasing concentrations of Nilotinib, Imatinib, Dasatinib and Ponatinib; in parallel 44 PBMC samples from 15 patients with steroid-dependent/refractory cGVHD treated with Nilotinib in the setting of a phase 1-2 trial were analyzed at baseline, after 90 and after 180 days of therapy. Flow cytometry was performed after labelling lymphocytes with a panel of monoclonal antibodies (CD3, CD4, CD16, CD56, CD25, CD19, CD45RA, FoxP3, CD127 and 7AAD). Cytokine production was assessed in supernatants of purified CD3+ T cells and in plasma samples from Nilotinib-treated patients. Main T lymphocytes subpopulations were not significantly affected by therapeutic concentrations of TKIs in vitro, whereas pro-inflammatory cytokine (in particular IL2, IFNγ, TNFα and IL10) and IL17 production showed a sharp decline. Frequency of T regulatory, B and NK cells decreased progressively in presence of therapeutic concentrations of all the TKIs tested in vitro, except for Nilotinib that showed little effect on these subsets; of note, na{\"i}ve T regulatory cell subset accumulated following exposure to TKIs. Results obtained in vivo on Nilotinib-treated patients were largely comparable, both on lymphocyte subset kinetics and on cytokine production by CD3-positive cells. This study underlines the anti-inflammatory and immunomodulatory effects of TKIs, and support their potential usefulness as treatment for patients with steroid dependent/refractory cGVHD. In addition, both in vitro and in vivo data point out that, compared to other TKIs, Nilotinib could better preserve the integrity of some important regulatory subsets, such as T regulatory and NK cells.",

keywords = "Tyrosine kinase inhibitors (TKIs), Chronic Graft Versus Host Disease (cGVHD), Lymphocyte subpopulations, T regulatory cells, Cytokine production",

author = "{MARINELLI BUSILACCHI}, Elena and Andrea COSTANTINI and Nadia VIOLA and Benedetta COSTANTINI and Jacopo OLIVIERI and Luca BUTINI and Giorgia MANCINI and Ilaria SCORTECHINI and Martina CHIARUCCI and Monica POIANI and Antonella POLONI and Pietro LEONI and Attilio OLIVIERI",

year = "2017",

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day = "8",

doi = "10.1016/j.bbmt.2017.10.039",

language = "English",

journal = "BIOLOGY OF BLOOD AND MARROW TRANSPLANTATION",

issn = "1083-8791",

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T1 - Immunomodulatory Effects of Tyrosine Kinase Inhibitors (TKIs) in Vitro and in Vivo Study

AU - MARINELLI BUSILACCHI, Elena

AU - COSTANTINI, Andrea

AU - VIOLA, Nadia

AU - COSTANTINI, Benedetta

AU - OLIVIERI, Jacopo

AU - BUTINI, Luca

AU - MANCINI, Giorgia

AU - SCORTECHINI, Ilaria

AU - CHIARUCCI, Martina

AU - POIANI, Monica

AU - POLONI, Antonella

AU - LEONI, Pietro

AU - OLIVIERI, Attilio

PY - 2017/11/8

Y1 - 2017/11/8

N2 - Pathogenesis of chronic Graft Versus Host Disease (cGVHD) is incompletely defined, involving donor-derived CD4 and CD8-positive T lymphocytes, as well as B cells. Standard treatment is lacking for steroid dependent/refractory cases; potential usefulness of tyrosine kinase inhibitors (TKIs) has been suggested, based on their potent anti-fibrotic effect. However, TKI seems to have pleiotropic activity. We sought to evaluate the in vitro and in vivo impact of different TKIs on lymphocyte phenotype and function. Peripheral blood mononuclear cells (PBMC) from healthy donors were cultured in presence of increasing concentrations of Nilotinib, Imatinib, Dasatinib and Ponatinib; in parallel 44 PBMC samples from 15 patients with steroid-dependent/refractory cGVHD treated with Nilotinib in the setting of a phase 1-2 trial were analyzed at baseline, after 90 and after 180 days of therapy. Flow cytometry was performed after labelling lymphocytes with a panel of monoclonal antibodies (CD3, CD4, CD16, CD56, CD25, CD19, CD45RA, FoxP3, CD127 and 7AAD). Cytokine production was assessed in supernatants of purified CD3+ T cells and in plasma samples from Nilotinib-treated patients. Main T lymphocytes subpopulations were not significantly affected by therapeutic concentrations of TKIs in vitro, whereas pro-inflammatory cytokine (in particular IL2, IFNγ, TNFα and IL10) and IL17 production showed a sharp decline. Frequency of T regulatory, B and NK cells decreased progressively in presence of therapeutic concentrations of all the TKIs tested in vitro, except for Nilotinib that showed little effect on these subsets; of note, naïve T regulatory cell subset accumulated following exposure to TKIs. Results obtained in vivo on Nilotinib-treated patients were largely comparable, both on lymphocyte subset kinetics and on cytokine production by CD3-positive cells. This study underlines the anti-inflammatory and immunomodulatory effects of TKIs, and support their potential usefulness as treatment for patients with steroid dependent/refractory cGVHD. In addition, both in vitro and in vivo data point out that, compared to other TKIs, Nilotinib could better preserve the integrity of some important regulatory subsets, such as T regulatory and NK cells.

AB - Pathogenesis of chronic Graft Versus Host Disease (cGVHD) is incompletely defined, involving donor-derived CD4 and CD8-positive T lymphocytes, as well as B cells. Standard treatment is lacking for steroid dependent/refractory cases; potential usefulness of tyrosine kinase inhibitors (TKIs) has been suggested, based on their potent anti-fibrotic effect. However, TKI seems to have pleiotropic activity. We sought to evaluate the in vitro and in vivo impact of different TKIs on lymphocyte phenotype and function. Peripheral blood mononuclear cells (PBMC) from healthy donors were cultured in presence of increasing concentrations of Nilotinib, Imatinib, Dasatinib and Ponatinib; in parallel 44 PBMC samples from 15 patients with steroid-dependent/refractory cGVHD treated with Nilotinib in the setting of a phase 1-2 trial were analyzed at baseline, after 90 and after 180 days of therapy. Flow cytometry was performed after labelling lymphocytes with a panel of monoclonal antibodies (CD3, CD4, CD16, CD56, CD25, CD19, CD45RA, FoxP3, CD127 and 7AAD). Cytokine production was assessed in supernatants of purified CD3+ T cells and in plasma samples from Nilotinib-treated patients. Main T lymphocytes subpopulations were not significantly affected by therapeutic concentrations of TKIs in vitro, whereas pro-inflammatory cytokine (in particular IL2, IFNγ, TNFα and IL10) and IL17 production showed a sharp decline. Frequency of T regulatory, B and NK cells decreased progressively in presence of therapeutic concentrations of all the TKIs tested in vitro, except for Nilotinib that showed little effect on these subsets; of note, naïve T regulatory cell subset accumulated following exposure to TKIs. Results obtained in vivo on Nilotinib-treated patients were largely comparable, both on lymphocyte subset kinetics and on cytokine production by CD3-positive cells. This study underlines the anti-inflammatory and immunomodulatory effects of TKIs, and support their potential usefulness as treatment for patients with steroid dependent/refractory cGVHD. In addition, both in vitro and in vivo data point out that, compared to other TKIs, Nilotinib could better preserve the integrity of some important regulatory subsets, such as T regulatory and NK cells.

KW - Tyrosine kinase inhibitors (TKIs)

KW - Chronic Graft Versus Host Disease (cGVHD)

KW - Lymphocyte subpopulations

KW - T regulatory cells

KW - Cytokine production

U2 - 10.1016/j.bbmt.2017.10.039

DO - 10.1016/j.bbmt.2017.10.039

M3 - Article

SN - 1083-8791

JO - BIOLOGY OF BLOOD AND MARROW TRANSPLANTATION

JF - BIOLOGY OF BLOOD AND MARROW TRANSPLANTATION

ER -

Immunomodulatory Effects of Tyrosine Kinase Inhibitors (TKIs) in Vitro and in Vivo Study

Abstract

Keywords

UN SDGs

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