Combined Vorinostat and Chloroquine Inhibit Sodium Iodide Symporter Endocytosis and Enhance Radionuclide Uptake In Vivo

Martin Read; Katie Brookes; Ling Zha; Selvambigai Manivannan; Jana Kim; Merve Kocbiyik; Alice Fletcher; Caroline M Gorvin; George Firth; Gilbert Fruhwirth; Juan Nicola; Sissy Jhiang; Matthew Ringel; Moray Campbell; Kavitha Sunassee; Philip Blower; Kristien Boelaert; Hannah Nieto; Vicki Smith; Christopher McCabe

doi:10.1158/1078-0432.CCR-23-2043

Combined Vorinostat and Chloroquine Inhibit Sodium Iodide Symporter Endocytosis and Enhance Radionuclide Uptake In Vivo

Martin Read, Katie Brookes, Ling Zha, Selvambigai Manivannan, Jana Kim, Merve Kocbiyik, Alice Fletcher, Caroline M Gorvin, George Firth, Gilbert Fruhwirth, Juan Nicola, Sissy Jhiang, Matthew Ringel, Moray Campbell, Kavitha Sunassee, Philip Blower, Kristien Boelaert, Hannah Nieto, Vicki Smith, Christopher McCabe^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

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Abstract

Purpose
Patients with aggressive thyroid cancer are frequently failed by the central therapy of ablative radioiodide (RAI) uptake, due to reduced plasma membrane (PM) localization of the sodium/iodide symporter (NIS). We aimed to understand how NIS is endocytosed away from the PM of human thyroid cancer cells, and whether this was druggable in vivo.

Experimental Design
Informed by analysis of endocytic gene expression in patients with aggressive thyroid cancer, we used mutagenesis, NanoBiT interaction assays, cell surface biotinylation assays, RAI uptake and NanoBRET to understand the mechanisms of NIS endocytosis in transformed cell lines and patient-derived human primary thyroid cells. Systemic drug responses were monitored via 99mTc pertechnetate gamma counting and gene expression in BALB/c mice.

Results
We identify an acidic dipeptide within the NIS C-terminus which mediates binding to the 2 subunit of the Adaptor Protein 2 (AP2) heterotetramer. We discovered that the FDA-approved drug chloroquine modulates NIS accumulation at the PM in a functional manner that is AP2 dependent. In vivo, chloroquine treatment of BALB/c mice significantly enhanced thyroidal uptake of 99mTc pertechnetate in combination with the histone deacetylase (HDAC) inhibitor vorinostat/ SAHA, accompanied by increased thyroidal NIS mRNA. Bioinformatic analyses validated the clinical relevance of AP2 genes with disease-free survival in RAI-treated DTC, enabling construction of an AP2 gene-related risk score classifier for predicting recurrence.

Conclusions
NIS internalisation is specifically druggable in vivo. Our data therefore provide new translatable potential for improving RAI therapy using FDA-approved drugs in patients with aggressive thyroid cancer.

Original language	English
Journal	Clinical Cancer Research
Early online date	3 Nov 2023
DOIs	https://doi.org/10.1158/1078-0432.CCR-23-2043
Publication status	E-pub ahead of print - 3 Nov 2023

Bibliographical note

This work was supported by the Department of Defense (BC201532P1 to M.J. Campbell and
596 C.J. McCabe) and Wellcome Trust (RG_05-052 to A. Fletcher). C.J. McCabe also received
597 funding from the Medical Research Council (CiC/1001505) and British Thyroid Foundation
598 (1002175). We acknowledge the labs of K. Pfleger and N. Lambert for kindly providing Venus599 tagged constructs, J.E. Blower for in vivo expertise, and D.P. Larner and R.L. Hoare for
600 technical expertise. We further acknowledge support from the Wellcome Trust and EPSRC
601 funded Centre for Medical Engineering at King’s College London (203148/Z/16/Z), the
602 Wellcome Multiuser Equipment Radioanalytical Facility (212885/Z/18/Z), and the EPSRC
603 programme for Next Generation Molecular Imaging and Therapy with Radionuclides
604 (EP/S019901/1).

UN SDGs

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

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10.1158/1078-0432.CCR-23-2043Licence: None: All rights reserved

ReadML2023Combined_AAMAccepted author manuscript, 15.1 MBLicence: Creative Commons: Attribution (CC BY)

2 Finished
2 Active

Making Radioiodine Treatment a Realistic Therapeutic Opportunity in Breast Cancer
McCabe, C.
US ARMY
30/09/21 → 29/09/25
Project: Research
Can new drug approaches elicit a novel dual action of radioiodine treatment?
McCabe, C.
BRITISH THYROID FOUNDATION
1/09/21 → 31/05/24
Project: Research
MRC CiC New clotrimazole analogues to improve radioiodine treatment of thyroid cancer
McCabe, C., Cox, L. & Bottegoni, G.
Medical Research Council
1/06/21 → 31/03/23
Project: Research Councils
WT ISSF 3 - Identification of novel sodium iodide symporter interactors which modulate radioiodine uptake
McCabe, C.
THE WELLCOME TRUST
1/10/18 → 30/09/19
Project: Research

Cite this

Read, M., Brookes, K., Zha, L., Manivannan, S., Kim, J., Kocbiyik, M., Fletcher, A., Gorvin, C. M., Firth, G., Fruhwirth, G., Nicola, J., Jhiang, S., Ringel, M., Campbell, M., Sunassee, K., Blower, P., Boelaert, K., Nieto, H., Smith, V., & McCabe, C. (2023). Combined Vorinostat and Chloroquine Inhibit Sodium Iodide Symporter Endocytosis and Enhance Radionuclide Uptake In Vivo. Clinical Cancer Research. Advance online publication. https://doi.org/10.1158/1078-0432.CCR-23-2043

@article{84e481bf0d584bb1ac6fa7f466ce7ea9,

title = "Combined Vorinostat and Chloroquine Inhibit Sodium Iodide Symporter Endocytosis and Enhance Radionuclide Uptake In Vivo",

abstract = "Purpose Patients with aggressive thyroid cancer are frequently failed by the central therapy of ablative radioiodide (RAI) uptake, due to reduced plasma membrane (PM) localization of the sodium/iodide symporter (NIS). We aimed to understand how NIS is endocytosed away from the PM of human thyroid cancer cells, and whether this was druggable in vivo.Experimental DesignInformed by analysis of endocytic gene expression in patients with aggressive thyroid cancer, we used mutagenesis, NanoBiT interaction assays, cell surface biotinylation assays, RAI uptake and NanoBRET to understand the mechanisms of NIS endocytosis in transformed cell lines and patient-derived human primary thyroid cells. Systemic drug responses were monitored via 99mTc pertechnetate gamma counting and gene expression in BALB/c mice.ResultsWe identify an acidic dipeptide within the NIS C-terminus which mediates binding to the 2 subunit of the Adaptor Protein 2 (AP2) heterotetramer. We discovered that the FDA-approved drug chloroquine modulates NIS accumulation at the PM in a functional manner that is AP2 dependent. In vivo, chloroquine treatment of BALB/c mice significantly enhanced thyroidal uptake of 99mTc pertechnetate in combination with the histone deacetylase (HDAC) inhibitor vorinostat/ SAHA, accompanied by increased thyroidal NIS mRNA. Bioinformatic analyses validated the clinical relevance of AP2 genes with disease-free survival in RAI-treated DTC, enabling construction of an AP2 gene-related risk score classifier for predicting recurrence.ConclusionsNIS internalisation is specifically druggable in vivo. Our data therefore provide new translatable potential for improving RAI therapy using FDA-approved drugs in patients with aggressive thyroid cancer.",

author = "Martin Read and Katie Brookes and Ling Zha and Selvambigai Manivannan and Jana Kim and Merve Kocbiyik and Alice Fletcher and Gorvin, {Caroline M} and George Firth and Gilbert Fruhwirth and Juan Nicola and Sissy Jhiang and Matthew Ringel and Moray Campbell and Kavitha Sunassee and Philip Blower and Kristien Boelaert and Hannah Nieto and Vicki Smith and Christopher McCabe",

note = "This work was supported by the Department of Defense (BC201532P1 to M.J. Campbell and 596 C.J. McCabe) and Wellcome Trust (RG_05-052 to A. Fletcher). C.J. McCabe also received 597 funding from the Medical Research Council (CiC/1001505) and British Thyroid Foundation 598 (1002175). We acknowledge the labs of K. Pfleger and N. Lambert for kindly providing Venus599 tagged constructs, J.E. Blower for in vivo expertise, and D.P. Larner and R.L. Hoare for 600 technical expertise. We further acknowledge support from the Wellcome Trust and EPSRC 601 funded Centre for Medical Engineering at King{\textquoteright}s College London (203148/Z/16/Z), the 602 Wellcome Multiuser Equipment Radioanalytical Facility (212885/Z/18/Z), and the EPSRC 603 programme for Next Generation Molecular Imaging and Therapy with Radionuclides 604 (EP/S019901/1). ",

year = "2023",

month = nov,

day = "3",

doi = "10.1158/1078-0432.CCR-23-2043",

language = "English",

journal = "Clinical Cancer Research",

issn = "1078-0432",

publisher = "American Association for Cancer Research",

}

Read, M, Brookes, K, Zha, L, Manivannan, S, Kim, J, Kocbiyik, M, Fletcher, A, Gorvin, CM, Firth, G, Fruhwirth, G, Nicola, J, Jhiang, S, Ringel, M, Campbell, M, Sunassee, K, Blower, P, Boelaert, K , Nieto, H , Smith, V & McCabe, C 2023, 'Combined Vorinostat and Chloroquine Inhibit Sodium Iodide Symporter Endocytosis and Enhance Radionuclide Uptake In Vivo', Clinical Cancer Research. https://doi.org/10.1158/1078-0432.CCR-23-2043

TY - JOUR

T1 - Combined Vorinostat and Chloroquine Inhibit Sodium Iodide Symporter Endocytosis and Enhance Radionuclide Uptake In Vivo

AU - Read, Martin

AU - Brookes, Katie

AU - Zha, Ling

AU - Manivannan, Selvambigai

AU - Kim, Jana

AU - Kocbiyik, Merve

AU - Fletcher, Alice

AU - Gorvin, Caroline M

AU - Firth, George

AU - Fruhwirth, Gilbert

AU - Nicola, Juan

AU - Jhiang, Sissy

AU - Ringel, Matthew

AU - Campbell, Moray

AU - Sunassee, Kavitha

AU - Blower, Philip

AU - Boelaert, Kristien

AU - Nieto, Hannah

AU - Smith, Vicki

AU - McCabe, Christopher

N1 - This work was supported by the Department of Defense (BC201532P1 to M.J. Campbell and 596 C.J. McCabe) and Wellcome Trust (RG_05-052 to A. Fletcher). C.J. McCabe also received 597 funding from the Medical Research Council (CiC/1001505) and British Thyroid Foundation 598 (1002175). We acknowledge the labs of K. Pfleger and N. Lambert for kindly providing Venus599 tagged constructs, J.E. Blower for in vivo expertise, and D.P. Larner and R.L. Hoare for 600 technical expertise. We further acknowledge support from the Wellcome Trust and EPSRC 601 funded Centre for Medical Engineering at King’s College London (203148/Z/16/Z), the 602 Wellcome Multiuser Equipment Radioanalytical Facility (212885/Z/18/Z), and the EPSRC 603 programme for Next Generation Molecular Imaging and Therapy with Radionuclides 604 (EP/S019901/1).

PY - 2023/11/3

Y1 - 2023/11/3

N2 - Purpose Patients with aggressive thyroid cancer are frequently failed by the central therapy of ablative radioiodide (RAI) uptake, due to reduced plasma membrane (PM) localization of the sodium/iodide symporter (NIS). We aimed to understand how NIS is endocytosed away from the PM of human thyroid cancer cells, and whether this was druggable in vivo.Experimental DesignInformed by analysis of endocytic gene expression in patients with aggressive thyroid cancer, we used mutagenesis, NanoBiT interaction assays, cell surface biotinylation assays, RAI uptake and NanoBRET to understand the mechanisms of NIS endocytosis in transformed cell lines and patient-derived human primary thyroid cells. Systemic drug responses were monitored via 99mTc pertechnetate gamma counting and gene expression in BALB/c mice.ResultsWe identify an acidic dipeptide within the NIS C-terminus which mediates binding to the 2 subunit of the Adaptor Protein 2 (AP2) heterotetramer. We discovered that the FDA-approved drug chloroquine modulates NIS accumulation at the PM in a functional manner that is AP2 dependent. In vivo, chloroquine treatment of BALB/c mice significantly enhanced thyroidal uptake of 99mTc pertechnetate in combination with the histone deacetylase (HDAC) inhibitor vorinostat/ SAHA, accompanied by increased thyroidal NIS mRNA. Bioinformatic analyses validated the clinical relevance of AP2 genes with disease-free survival in RAI-treated DTC, enabling construction of an AP2 gene-related risk score classifier for predicting recurrence.ConclusionsNIS internalisation is specifically druggable in vivo. Our data therefore provide new translatable potential for improving RAI therapy using FDA-approved drugs in patients with aggressive thyroid cancer.

AB - Purpose Patients with aggressive thyroid cancer are frequently failed by the central therapy of ablative radioiodide (RAI) uptake, due to reduced plasma membrane (PM) localization of the sodium/iodide symporter (NIS). We aimed to understand how NIS is endocytosed away from the PM of human thyroid cancer cells, and whether this was druggable in vivo.Experimental DesignInformed by analysis of endocytic gene expression in patients with aggressive thyroid cancer, we used mutagenesis, NanoBiT interaction assays, cell surface biotinylation assays, RAI uptake and NanoBRET to understand the mechanisms of NIS endocytosis in transformed cell lines and patient-derived human primary thyroid cells. Systemic drug responses were monitored via 99mTc pertechnetate gamma counting and gene expression in BALB/c mice.ResultsWe identify an acidic dipeptide within the NIS C-terminus which mediates binding to the 2 subunit of the Adaptor Protein 2 (AP2) heterotetramer. We discovered that the FDA-approved drug chloroquine modulates NIS accumulation at the PM in a functional manner that is AP2 dependent. In vivo, chloroquine treatment of BALB/c mice significantly enhanced thyroidal uptake of 99mTc pertechnetate in combination with the histone deacetylase (HDAC) inhibitor vorinostat/ SAHA, accompanied by increased thyroidal NIS mRNA. Bioinformatic analyses validated the clinical relevance of AP2 genes with disease-free survival in RAI-treated DTC, enabling construction of an AP2 gene-related risk score classifier for predicting recurrence.ConclusionsNIS internalisation is specifically druggable in vivo. Our data therefore provide new translatable potential for improving RAI therapy using FDA-approved drugs in patients with aggressive thyroid cancer.

U2 - 10.1158/1078-0432.CCR-23-2043

DO - 10.1158/1078-0432.CCR-23-2043

M3 - Article

SN - 1078-0432

JO - Clinical Cancer Research

JF - Clinical Cancer Research

ER -

Combined Vorinostat and Chloroquine Inhibit Sodium Iodide Symporter Endocytosis and Enhance Radionuclide Uptake In Vivo

Abstract

Bibliographical note

UN SDGs

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Projects

Making Radioiodine Treatment a Realistic Therapeutic Opportunity in Breast Cancer

Can new drug approaches elicit a novel dual action of radioiodine treatment?

MRC CiC New clotrimazole analogues to improve radioiodine treatment of thyroid cancer

WT ISSF 3 - Identification of novel sodium iodide symporter interactors which modulate radioiodine uptake

Cite this