A copper-based metabolite of disulfiram upregulates sodium iodide symporter (NIS) gene expression to enhance thyroidal uptake of radionuclides in vivo

Katie Brookes; Ling Zha; Benjamin Small; Jana Kim; Selvambigai Manivannan; Caitlin Thornton; Vinodh Kannappan; Weiguang Wang; Hannah Nieto; Kavitha Sunassee; Philip Blower; Vicki Smith; Martin Read; Christopher McCabe

doi:10.1530/endoabs.94.OC7.1

Abstract

Introduction: New approaches to improve radioiodide (RAI) uptake are urgently required in RAI-refractory thyroid cancer. We previously identified disulfiram as a leading candidate to induce sodium iodide symporter (NIS) activity and promote RAI uptake. In vivo, DSF is metabolised to diethyldithiocarbamate (DDC) which binds metal ions. Here, we aimed to gain a mechanistic understanding of how DSF and it’s related metabolite Cu(DDC)₂ impact NIS activity.

Methods: NIS function was monitored by RAI (¹²⁵I) uptake assays. Global gene expression changes in response to Cu(DDC)₂ were appraised via in vitro RNAseq analysis. Technetium-99m pertechnetate (^99mTc) uptake was used to evaluate NIS function in wild-type BALB/c mice.

Results: The ability of DSF to increase RAI uptake in TPC-1 (3.1-fold; P<0.01) and 8505C (4.9-fold; P<0.001) cells was potentiated by combination with Cu²⁺ to 5.1- and 18.9-fold increases, respectively. Similarly, Cu(DDC)₂ was highly effective at increasing RAI uptake (up to 8-fold;250nM; P<0.001) in multiple thyroid cell types and induced NIS protein expression, whilst methylated-DDC lacking Cu²⁺ had no effect. Interestingly, a potent transcriptional effect of Cu(DDC)₂ was revealed via NIS mRNA induction in TPC-1 (8.5-fold; P<0.001) and 8505C (104.8-fold; P<0.001) cells. RNA-Seq analysis of Cu(DDC)₂-treated 8505C cells revealed altered expression of NIS transcriptional regulators, including PAX8 (+4.02-fold; P=0.009), CREM (+1.97-fold; P=0.003), SMAD3 (-1.66-fold; P=0.012) and NKX2-1 (-1.93-fold; P=0.026). Intraperitoneal administration of Cu(DDC)₂ in wild-type BALB/c mice significantly induced thyroidal uptake of ^99mTc after 30 min (~40% increase;3mg/kg dose; P<0.001), as well as increasing thyroidal NIS (1.9-fold; P<0.01), thyroid peroxidase (1.8-fold; P<0.001) and thyroglobulin (1.3-fold; P<0.05) mRNA expression. Importantly, there was a significant positive correlation between thyroidal ^99mTc uptake and NIS mRNA levels (r_s=0.448, P=0.0169) in Cu(DDC)₂-treated mice.

Discussion: Our study demonstrates that a copper-disulfiram metabolite induces a transcriptional response to increase NIS activity in vitro and in vivo, with clinical potential to improve RAI-refractory thyroid cancer treatment.

Original language	English
Article number	OC7.1
Number of pages	1
Journal	Endocrine Abstracts
Volume	94
DOIs	https://doi.org/10.1530/endoabs.94.OC7.1
Publication status	Published - 30 Oct 2023
Event	Society for Endocrinology BES 2023 - SEC, Glasgow, United Kingdom Duration: 13 Nov 2023 → 15 Nov 2023

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Making Radioiodine Treatment a Realistic Therapeutic Opportunity in Breast Cancer
McCabe, C.
US ARMY
30/09/21 → 29/09/25
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Brookes, K., Zha, L., Small, B., Kim, J., Manivannan, S., Thornton, C., Kannappan, V., Wang, W., Nieto, H., Sunassee, K., Blower, P., Smith, V., Read, M., & McCabe, C. (2023). A copper-based metabolite of disulfiram upregulates sodium iodide symporter (NIS) gene expression to enhance thyroidal uptake of radionuclides in vivo. Endocrine Abstracts, 94, Article OC7.1. https://doi.org/10.1530/endoabs.94.OC7.1

@article{7c72e721a27349478e2356bb7b63d377,

title = "A copper-based metabolite of disulfiram upregulates sodium iodide symporter (NIS) gene expression to enhance thyroidal uptake of radionuclides in vivo",

abstract = "Introduction: New approaches to improve radioiodide (RAI) uptake are urgently required in RAI-refractory thyroid cancer. We previously identified disulfiram as a leading candidate to induce sodium iodide symporter (NIS) activity and promote RAI uptake. In vivo, DSF is metabolised to diethyldithiocarbamate (DDC) which binds metal ions. Here, we aimed to gain a mechanistic understanding of how DSF and it{\textquoteright}s related metabolite Cu(DDC)2 impact NIS activity.Methods: NIS function was monitored by RAI (125I) uptake assays. Global gene expression changes in response to Cu(DDC)2 were appraised via in vitro RNAseq analysis. Technetium-99m pertechnetate (99mTc) uptake was used to evaluate NIS function in wild-type BALB/c mice.Results: The ability of DSF to increase RAI uptake in TPC-1 (3.1-fold; P<0.01) and 8505C (4.9-fold; P<0.001) cells was potentiated by combination with Cu2+ to 5.1- and 18.9-fold increases, respectively. Similarly, Cu(DDC)2 was highly effective at increasing RAI uptake (up to 8-fold;250nM; P<0.001) in multiple thyroid cell types and induced NIS protein expression, whilst methylated-DDC lacking Cu2+ had no effect. Interestingly, a potent transcriptional effect of Cu(DDC)2 was revealed via NIS mRNA induction in TPC-1 (8.5-fold; P<0.001) and 8505C (104.8-fold; P<0.001) cells. RNA-Seq analysis of Cu(DDC)2-treated 8505C cells revealed altered expression of NIS transcriptional regulators, including PAX8 (+4.02-fold; P=0.009), CREM (+1.97-fold; P=0.003), SMAD3 (-1.66-fold; P=0.012) and NKX2-1 (-1.93-fold; P=0.026). Intraperitoneal administration of Cu(DDC)2 in wild-type BALB/c mice significantly induced thyroidal uptake of 99mTc after 30 min (~40% increase;3mg/kg dose; P<0.001), as well as increasing thyroidal NIS (1.9-fold; P<0.01), thyroid peroxidase (1.8-fold; P<0.001) and thyroglobulin (1.3-fold; P<0.05) mRNA expression. Importantly, there was a significant positive correlation between thyroidal 99mTc uptake and NIS mRNA levels (rs=0.448, P=0.0169) in Cu(DDC)2-treated mice.Discussion: Our study demonstrates that a copper-disulfiram metabolite induces a transcriptional response to increase NIS activity in vitro and in vivo, with clinical potential to improve RAI-refractory thyroid cancer treatment.",

author = "Katie Brookes and Ling Zha and Benjamin Small and Jana Kim and Selvambigai Manivannan and Caitlin Thornton and Vinodh Kannappan and Weiguang Wang and Hannah Nieto and Kavitha Sunassee and Philip Blower and Vicki Smith and Martin Read and Christopher McCabe",

year = "2023",

month = oct,

day = "30",

doi = "10.1530/endoabs.94.OC7.1",

language = "English",

volume = "94",

journal = "Endocrine Abstracts",

issn = "1470-3947",

publisher = "BioScientifica",

note = "Society for Endocrinology BES 2023 ; Conference date: 13-11-2023 Through 15-11-2023",

}

Brookes, K, Zha, L, Small, B, Kim, J, Manivannan, S, Thornton, C, Kannappan, V, Wang, W, Nieto, H, Sunassee, K, Blower, P, Smith, V , Read, M & McCabe, C 2023, 'A copper-based metabolite of disulfiram upregulates sodium iodide symporter (NIS) gene expression to enhance thyroidal uptake of radionuclides in vivo', Endocrine Abstracts, vol. 94, OC7.1. https://doi.org/10.1530/endoabs.94.OC7.1

TY - JOUR

T1 - A copper-based metabolite of disulfiram upregulates sodium iodide symporter (NIS) gene expression to enhance thyroidal uptake of radionuclides in vivo

AU - Brookes, Katie

AU - Zha, Ling

AU - Small, Benjamin

AU - Kim, Jana

AU - Manivannan, Selvambigai

AU - Thornton, Caitlin

AU - Kannappan, Vinodh

AU - Wang, Weiguang

AU - Nieto, Hannah

AU - Sunassee, Kavitha

AU - Blower, Philip

AU - Smith, Vicki

AU - Read, Martin

AU - McCabe, Christopher

PY - 2023/10/30

Y1 - 2023/10/30

N2 - Introduction: New approaches to improve radioiodide (RAI) uptake are urgently required in RAI-refractory thyroid cancer. We previously identified disulfiram as a leading candidate to induce sodium iodide symporter (NIS) activity and promote RAI uptake. In vivo, DSF is metabolised to diethyldithiocarbamate (DDC) which binds metal ions. Here, we aimed to gain a mechanistic understanding of how DSF and it’s related metabolite Cu(DDC)2 impact NIS activity.Methods: NIS function was monitored by RAI (125I) uptake assays. Global gene expression changes in response to Cu(DDC)2 were appraised via in vitro RNAseq analysis. Technetium-99m pertechnetate (99mTc) uptake was used to evaluate NIS function in wild-type BALB/c mice.Results: The ability of DSF to increase RAI uptake in TPC-1 (3.1-fold; P<0.01) and 8505C (4.9-fold; P<0.001) cells was potentiated by combination with Cu2+ to 5.1- and 18.9-fold increases, respectively. Similarly, Cu(DDC)2 was highly effective at increasing RAI uptake (up to 8-fold;250nM; P<0.001) in multiple thyroid cell types and induced NIS protein expression, whilst methylated-DDC lacking Cu2+ had no effect. Interestingly, a potent transcriptional effect of Cu(DDC)2 was revealed via NIS mRNA induction in TPC-1 (8.5-fold; P<0.001) and 8505C (104.8-fold; P<0.001) cells. RNA-Seq analysis of Cu(DDC)2-treated 8505C cells revealed altered expression of NIS transcriptional regulators, including PAX8 (+4.02-fold; P=0.009), CREM (+1.97-fold; P=0.003), SMAD3 (-1.66-fold; P=0.012) and NKX2-1 (-1.93-fold; P=0.026). Intraperitoneal administration of Cu(DDC)2 in wild-type BALB/c mice significantly induced thyroidal uptake of 99mTc after 30 min (~40% increase;3mg/kg dose; P<0.001), as well as increasing thyroidal NIS (1.9-fold; P<0.01), thyroid peroxidase (1.8-fold; P<0.001) and thyroglobulin (1.3-fold; P<0.05) mRNA expression. Importantly, there was a significant positive correlation between thyroidal 99mTc uptake and NIS mRNA levels (rs=0.448, P=0.0169) in Cu(DDC)2-treated mice.Discussion: Our study demonstrates that a copper-disulfiram metabolite induces a transcriptional response to increase NIS activity in vitro and in vivo, with clinical potential to improve RAI-refractory thyroid cancer treatment.

AB - Introduction: New approaches to improve radioiodide (RAI) uptake are urgently required in RAI-refractory thyroid cancer. We previously identified disulfiram as a leading candidate to induce sodium iodide symporter (NIS) activity and promote RAI uptake. In vivo, DSF is metabolised to diethyldithiocarbamate (DDC) which binds metal ions. Here, we aimed to gain a mechanistic understanding of how DSF and it’s related metabolite Cu(DDC)2 impact NIS activity.Methods: NIS function was monitored by RAI (125I) uptake assays. Global gene expression changes in response to Cu(DDC)2 were appraised via in vitro RNAseq analysis. Technetium-99m pertechnetate (99mTc) uptake was used to evaluate NIS function in wild-type BALB/c mice.Results: The ability of DSF to increase RAI uptake in TPC-1 (3.1-fold; P<0.01) and 8505C (4.9-fold; P<0.001) cells was potentiated by combination with Cu2+ to 5.1- and 18.9-fold increases, respectively. Similarly, Cu(DDC)2 was highly effective at increasing RAI uptake (up to 8-fold;250nM; P<0.001) in multiple thyroid cell types and induced NIS protein expression, whilst methylated-DDC lacking Cu2+ had no effect. Interestingly, a potent transcriptional effect of Cu(DDC)2 was revealed via NIS mRNA induction in TPC-1 (8.5-fold; P<0.001) and 8505C (104.8-fold; P<0.001) cells. RNA-Seq analysis of Cu(DDC)2-treated 8505C cells revealed altered expression of NIS transcriptional regulators, including PAX8 (+4.02-fold; P=0.009), CREM (+1.97-fold; P=0.003), SMAD3 (-1.66-fold; P=0.012) and NKX2-1 (-1.93-fold; P=0.026). Intraperitoneal administration of Cu(DDC)2 in wild-type BALB/c mice significantly induced thyroidal uptake of 99mTc after 30 min (~40% increase;3mg/kg dose; P<0.001), as well as increasing thyroidal NIS (1.9-fold; P<0.01), thyroid peroxidase (1.8-fold; P<0.001) and thyroglobulin (1.3-fold; P<0.05) mRNA expression. Importantly, there was a significant positive correlation between thyroidal 99mTc uptake and NIS mRNA levels (rs=0.448, P=0.0169) in Cu(DDC)2-treated mice.Discussion: Our study demonstrates that a copper-disulfiram metabolite induces a transcriptional response to increase NIS activity in vitro and in vivo, with clinical potential to improve RAI-refractory thyroid cancer treatment.

U2 - 10.1530/endoabs.94.OC7.1

DO - 10.1530/endoabs.94.OC7.1

M3 - Abstract

SN - 1470-3947

VL - 94

JO - Endocrine Abstracts

JF - Endocrine Abstracts

M1 - OC7.1

T2 - Society for Endocrinology BES 2023

Y2 - 13 November 2023 through 15 November 2023

ER -

A copper-based metabolite of disulfiram upregulates sodium iodide symporter (NIS) gene expression to enhance thyroidal uptake of radionuclides in vivo

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Making Radioiodine Treatment a Realistic Therapeutic Opportunity in Breast Cancer

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