Skip to main content
  • AACR Journals
    • Cancer Discovery
    • Cancer Epidemiology, Biomarkers & Prevention
    • Cancer Immunology Research
    • Cancer Prevention Research
    • Cancer Research
    • Clinical Cancer Research
    • Molecular Cancer Research
    • Molecular Cancer Therapeutics

  • Register
  • Log in
Advertisement

Main menu

  • Home
  • About
    • The Journal
    • AACR Journals
    • Subscriptions
    • Permissions and Reprints
    • Reviewing
  • Articles
    • OnlineFirst
    • Current Issue
    • Past Issues
    • Meeting Abstracts
    • Molecular Targets Collections
  • For Authors
    • Information for Authors
    • Author Services
    • Best of: Author Profiles
    • Submit
  • Alerts
    • Table of Contents
    • OnlineFirst
    • Editors' Picks
    • Citation
    • Author/Keyword
  • News
    • Cancer Discovery News
  • AACR Journals
    • Cancer Discovery
    • Cancer Epidemiology, Biomarkers & Prevention
    • Cancer Immunology Research
    • Cancer Prevention Research
    • Cancer Research
    • Clinical Cancer Research
    • Molecular Cancer Research
    • Molecular Cancer Therapeutics

User menu

  • Register
  • Log in

Search

  • Advanced search
Molecular Cancer Therapeutics
Molecular Cancer Therapeutics

Advanced Search

  • Home
  • About
    • The Journal
    • AACR Journals
    • Subscriptions
    • Permissions and Reprints
    • Reviewing
  • Articles
    • OnlineFirst
    • Current Issue
    • Past Issues
    • Meeting Abstracts
    • Molecular Targets Collections
  • For Authors
    • Information for Authors
    • Author Services
    • Best of: Author Profiles
    • Submit
  • Alerts
    • Table of Contents
    • OnlineFirst
    • Editors' Picks
    • Citation
    • Author/Keyword
  • News
    • Cancer Discovery News
Small Molecule Therapeutics

Clinical Dosing Regimen of Selinexor Maintains Normal Immune Homeostasis and T-cell Effector Function in Mice: Implications for Combination with Immunotherapy

Paul M. Tyler, Mariah M. Servos, Romy C. de Vries, Boris Klebanov, Trinayan Kashyap, Sharon Sacham, Yosef Landesman, Michael Dougan and Stephanie K. Dougan
Paul M. Tyler
Dana-Farber Cancer Institute, Boston, Massachusetts.
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Mariah M. Servos
Dana-Farber Cancer Institute, Boston, Massachusetts.
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Romy C. de Vries
Dana-Farber Cancer Institute, Boston, Massachusetts.University of Amsterdam, Program in Biomedical Sciences, Amsterdam, the Netherlands.
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Boris Klebanov
Karyopharm Therapeutics, Inc., Newton, Massachusetts.
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Trinayan Kashyap
Karyopharm Therapeutics, Inc., Newton, Massachusetts.
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Sharon Sacham
Karyopharm Therapeutics, Inc., Newton, Massachusetts.
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Yosef Landesman
Karyopharm Therapeutics, Inc., Newton, Massachusetts.
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Michael Dougan
Massachusetts General Hospital, Boston, Massachusetts.
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Stephanie K. Dougan
Dana-Farber Cancer Institute, Boston, Massachusetts.
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • For correspondence: Stephanie_Dougan@dfci.harvard.edu
DOI: 10.1158/1535-7163.MCT-16-0496 Published March 2017
  • Article
  • Figures & Data
  • Info & Metrics
  • PDF
Loading

Article Figures & Data

Figures

  • Additional Files
  • Figure 1.
    • Download figure
    • Open in new tab
    • Download powerpoint
    Figure 1.

    Selinexor disrupts normal immune homeostasis. A, C57BL/6 mice were treated with vehicle or KPT-330 (15 mg/kg) by oral gavage 3 times per week (Monday, Wednesday, and Friday) for 2 or 4 weeks. Total thymocyte cell numbers are shown. n = 10, vehicle; n = 5 KPT-330 groups. B, Thymocytes from vehicle or 14-day treated mice in A were stained with antibodies to CD4 and CD8 and analyzed by flow cytometry. Representative image is shown. C, Quantification of thymic populations from vehicle or 14-day treated mice. D, Thymocytes from vehicle or 14-day treated mice in A were stained with antibodies to CD4, CD8, CD25, and CD44 and analyzed by flow cytometry. Representative image is shown, gated on CD4−CD8− cells. E, C57BL/6 mice were treated with vehicle or KPT-330 (15 mg/kg) by oral gavage 3 times per week (Monday, Wednesday, and Friday) for 4 weeks. Bone marrow cells were harvested, stained with antibodies, and analyzed by flow cytometry. CD8 T cells = CD45+CD8+; CD4 T cells = CD45+CD4+; myeloid = CD45+CD11b+Gr1−; pre, proB = CD45+B220+IgD−; immature/mature B = CD45+B220+IgD+; NK cells = CD45+NK1.1+; neutrophils = CD45+CD11b+Gr1+; DN T cells = CD45+CD3+CD4−CD8−; lineage− = CD45+ negative for all other markers. Pie chart shows average frequencies from n = 5 mice per group. F, C57BL/6 mice were treated with vehicle or KPT-330 (15 mg/kg) by oral gavage 3 times per week (Monday, Wednesday, and Friday) for 1, 2, or 4 weeks. Spleen cells were harvested, stained with antibodies, and analyzed by flow cytometry. B cells = CD45+B220+; CD4 T cells = CD45+CD4+Foxp3−; Tregs = CD45+CD4+Foxp3+; macrophages = CD45+CD11b+Gr1−; CD8 T cells = CD45+CD8+; NK cells = CD45+NK1.1+; monocytes = CD45+CD11b+Ly6C+Gr1−; neutrophils = CD45+CD11b+Gr1+. Pie charts show averages from n = 5 mice per group.

  • Figure 2.
    • Download figure
    • Open in new tab
    • Download powerpoint
    Figure 2.

    Antibody production is only modestly affected by selinexor. A, Diagram of KPT-330 dosing and immunization schedule. M, Monday; W, Wednesday; F, Friday. B, C57BL/6 mice were immunized according to diagram shown in A. Serum was collected at the indicated time points, diluted at the indicated ratios, and used in an ELISA with ovalbumin-coated plates. Secondary anti-mouse Igκ coupled to HRP was used for detection. Each line represents one mouse. n = 3 mice per group. C, Serum was analyzed as in B, using secondary anti-mouse IgG1, anti-mouse IgG2b, or anti-mouse IgG2c coupled to HRP for detection.

  • Figure 3.
    • Download figure
    • Open in new tab
    • Download powerpoint
    Figure 3.

    Decreased frequency of selinexor treatment restores immune homeostasis better than decreased dose. A, C57BL/6 mice were dosed for 3 weeks according to the indicated schedules. n = 5 mice per group. M, Monday; W, Wednesday; F, Friday. B, Total thymocyte cell numbers. C, Bone marrow cells were harvested and stained as in Fig. 1E. CD8 T cells and immature B cells were quantified. D, Mesenteric lymph node cells were harvested and stained as in Fig. 1F. Monocytes were quantified. E, Spleen cells were harvested and stained as in Fig. 1F. Monocytes and CD8 T cells were quantified.

  • Figure 4.
    • Download figure
    • Open in new tab
    • Download powerpoint
    Figure 4.

    Selinexor blocks activation of naïve T cells. A, CD8 T cells from an OT-I mouse were labeled with CellTrace violet and cocultured with CD40-activated B cells pulsed with ovalbumin peptide (SIINFEKL) at the indicated concentrations in the presence or absence of 1 μmol/L KPT-330. T-cell proliferation was measured by flow cytometry 3 days later. B, CD8 T cells from C57BL/6 mice were labeled with CellTrace violet and cocultured with anti-CD3/CD28–coated beads or PMA/Ion in the presence or absence of 1 μmol/L KPT-330. Three days later, cells were stained with anti-CD25 and anti-CD8 and analyzed by flow cytometry. C, Total lymph node cells from C57BL/6 mice were cultured with anti-CD3/CD28 beads and the indicated concentrations of KPT-330 added at the indicated time points relative to CD3/CD28 stimulation. Twenty-four hours later, cells were stained with antibodies to CD4, CD8, and CD25 and analyzed by flow cytometry.

  • Figure 5.
    • Download figure
    • Open in new tab
    • Download powerpoint
    Figure 5.

    Selinexor blocks the activation of effector CD8 T cells. A, CD8 T cells from an OT-I mouse were cultured with anti-CD3/CD28 beads for 7 days to generate effector cells. Effector cells were washed, counted, and plated onto B16OVA tumor cells that had been pretreated with IFNγ to induce MHC class I expression. KPT-330 was added to the cocultures at the indicated concentrations. Twenty-four hours later, OT-I cells were collected, fixed and permeabilized, and stained with antibodies to CD25, CD69, IFNγ, and granzyme B, and analyzed by flow cytometry. No response was seen from OT-I effector cells cultured with B16 cells not transduced with ovalbumin. Representative of three independent experiments. B, C57BL/6 mice were dosed with KPT-330 10 mg/kg by oral gavage. Serum was collected at various time points and analyzed for KPT-330 concentration by mass spectrometry. Dashed line, 100 nmol/L concentration.

  • Figure 6.
    • Download figure
    • Open in new tab
    • Download powerpoint
    Figure 6.

    Optimized selinexor dosing does not inhibit antitumor immunity in vivo. A, C57BL/6 mice were inoculated subcutaneously with 1 × 105 B16 cells on day 0. On days 7, 9, 14, and 16, mice were dosed with vehicle or 10 mg/kg KPT-330 by oral gavage. Tumors were harvested on day 21, digested and infiltrates analyzed by flow cytometry. Total CD45+ cells were quantified. n = 5 mice per group. Representative of two experiments. B, Tumor infiltrates from A were stained with antibodies to CD45, CD8, CD4, NK1.1, B220, MHC class II, Gr1, Foxp3, CD11b, and CD11c and analyzed by flow cytometry. n = 5 mice per group. Representative of two experiments. M, Monday; W, Wednesday. C, C57BL/6 mice were inoculated with B16 tumors and dosed with 10 mg/kg KPT-330 on days 6, 8, 13, and 15. On day 12 after tumor inoculation, all mice received adoptive transfer of 5 × 105 activated TRP1high;CD45.1+ CD8 T cells. On day 17, tumor infiltrates were harvested, stained with the indicated antibodies, and analyzed by flow cytometry after gating on CD45+CD8+CD45.1+ TRP1-specific cells. n = 5 mice per group. D, Tumor infiltrates from C were gated on endogenous CD8 T cells (CD45+CD8+CD45.1−). E, Tumor infiltrates from C were stained with antibodies to PD-L1 and other cell surface markers. Mean fluorescence intensity (MFI) of PD-L1 expression was quantified. Tumor = CD45−; Mac = CD45+CD11b+Gr1−CD11c−; MDSCs = CD45+CD11b+Gr1+; dendritic cells = CD45+MHCII+CD11c+. n = 5 mice per group. F, Tumor growth of mice analyzed in C and D.

Additional Files

  • Figures
  • Supplementary Data

    • Supplemental Figure 1 - Supplemental Figure 1: Response to immunization is only modestly affected by selinexor.
    • Supplemental Figure 2 - Supplemental Figure 2: IFNγ secretion is impaired by selinexor.
    • Supplemental Figure 3 - Supplemental Figure 3: Degranulation of effector CD8 T cells is affected by selinexor.
    • Supplemental Figure legends - Legends for supplemental figures
PreviousNext
Back to top
Molecular Cancer Therapeutics: 16 (3)
March 2017
Volume 16, Issue 3
  • Table of Contents
  • Table of Contents (PDF)
  • About the Cover
  • Index by Author
  • Editorial Board (PDF)

Sign up for alerts

View this article with LENS

Open full page PDF
Article Alerts
Sign In to Email Alerts with your Email Address
Email Article

Thank you for sharing this Molecular Cancer Therapeutics article.

NOTE: We request your email address only to inform the recipient that it was you who recommended this article, and that it is not junk mail. We do not retain these email addresses.

Enter multiple addresses on separate lines or separate them with commas.
Clinical Dosing Regimen of Selinexor Maintains Normal Immune Homeostasis and T-cell Effector Function in Mice: Implications for Combination with Immunotherapy
(Your Name) has forwarded a page to you from Molecular Cancer Therapeutics
(Your Name) thought you would be interested in this article in Molecular Cancer Therapeutics.
Citation Tools
Clinical Dosing Regimen of Selinexor Maintains Normal Immune Homeostasis and T-cell Effector Function in Mice: Implications for Combination with Immunotherapy
Paul M. Tyler, Mariah M. Servos, Romy C. de Vries, Boris Klebanov, Trinayan Kashyap, Sharon Sacham, Yosef Landesman, Michael Dougan and Stephanie K. Dougan
Mol Cancer Ther March 1 2017 (16) (3) 428-439; DOI: 10.1158/1535-7163.MCT-16-0496

Citation Manager Formats

  • BibTeX
  • Bookends
  • EasyBib
  • EndNote (tagged)
  • EndNote 8 (xml)
  • Medlars
  • Mendeley
  • Papers
  • RefWorks Tagged
  • Ref Manager
  • RIS
  • Zotero
Share
Clinical Dosing Regimen of Selinexor Maintains Normal Immune Homeostasis and T-cell Effector Function in Mice: Implications for Combination with Immunotherapy
Paul M. Tyler, Mariah M. Servos, Romy C. de Vries, Boris Klebanov, Trinayan Kashyap, Sharon Sacham, Yosef Landesman, Michael Dougan and Stephanie K. Dougan
Mol Cancer Ther March 1 2017 (16) (3) 428-439; DOI: 10.1158/1535-7163.MCT-16-0496
del.icio.us logo Digg logo Reddit logo Twitter logo CiteULike logo Facebook logo Google logo Mendeley logo
  • Tweet Widget
  • Facebook Like
  • Google Plus One

Jump to section

  • Article
    • Abstract
    • Introduction
    • Materials and Methods
    • Results
    • Discussion
    • Disclosure of Potential Conflicts of Interest
    • Authors' Contributions
    • Grant Support
    • Acknowledgments
    • Footnotes
    • References
  • Figures & Data
  • Info & Metrics
  • PDF
Advertisement

Related Articles

Cited By...

More in this TOC Section

  • Global Analysis of NT157 Action
  • A Novel Small RNA against KRAS-Mutant Colon Cancer
  • Trastuzumab-Resistant Breast Cancers Are Sensitive to PARPi
Show more Small Molecule Therapeutics
  • Home
  • Alerts
  • Feedback
Facebook  Twitter  LinkedIn  YouTube  RSS

Articles

  • Online First
  • Current Issue
  • Past Issues
  • Meeting Abstracts

Info for

  • Authors
  • Subscribers
  • Advertisers
  • Librarians
  • Reviewers

About MCT

  • About the Journal
  • Editorial Board
  • Permissions
  • Submit a Manuscript
AACR logo

Copyright 2016 by the American Association for Cancer Research.

Molecular Cancer Therapeutics
eISSN: 1538-8514
ISSN: 1535-7163

Advertisement