Supported by a grant from The Memorial Foundation of Manufacturer Vilhelm Pedersen &#38; Wife. The foundation played no role in the planning or any other phase of the study.

A few clinical trials using this 64Cu PET/CT or PET/MRI protocol have been approved by the Regional Ethics Committee of&#160;Region Midt, Denmark [1-10-72-196-16 (EudraCT 2016-001975-59), 1-10-72-41-19 (EudraCT 2019-000905-57), 1-10-72-343-20 (EudraCT 2020-005832-31), 1-10-72-25-21 (EudraCT 2021-000102-25), and 1-10-72-15-22 (EudraCT 2021-005464-21)]. Written informed consent was obtained from the participants at enrollment. The inclusion criteria for all participants were age &#62;18, and for females the use ...

<ol>
	<li>T&#252;mer, Z., M&#248;ller, L. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Menkes+disease.">Menkes disease.</a> European Journal of Human Genetics. 18 (5), 511-518 (2010).</li><li>Ala, A., Walker, A. P., Ashkan, K., Dooley, J. S., Schilsky, M. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Wilson's+disease.">Wilson's disease.</a> The Lancet. 369 (9559), 397-408 (2007).</li><li>Owen, C. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Absorption+and+excretion+of+Cu64-labeled+copper+by+the+rat.">Absorption and excretion of Cu64-labeled copper by the rat.</a> The American Journal of Physiology-Legacy Content. 207 (6), 1203-1206 (1964).</li><li>Osborn, S. B., Roberts, C. N., Walshe, J. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Uptake+of+radiocopper+by+the+liver.+A+study+of+patients+with+Wilson's+disease+and+various+control+groups.">Uptake of radiocopper by the liver. A study of patients with Wilson's disease and various control groups.</a> Clinical Science. 24, 13-22 (1963).</li><li>Vierling, J. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Incorporation+of+radiocopper+into+ceruloplasmin+in+normal+subjects+and+in+patients+with+primary+biliary+cirrhosis+and+Wilson's+disease.">Incorporation of radiocopper into ceruloplasmin in normal subjects and in patients with primary biliary cirrhosis and Wilson's disease.</a> Gastroenterology. 74 (4), 652-660 (1978).</li><li>Gibbs, K., Walshe, J. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Studies+with+radioactive+copper+(64Cu+and+67Cu);+the+incorporation+of+radioactive+copper+into+caeruloplasmin+in+Wilson's+disease+and+in+primary+biliary+cirrhosis.">Studies with radioactive copper (64Cu and 67Cu); the incorporation of radioactive copper into caeruloplasmin in Wilson's disease and in primary biliary cirrhosis.</a> Clinical Science. 41 (3), 189-202 (1971).</li><li>Kume, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+semi-automated+system+for+the+routine+production+of+copper-64.">A semi-automated system for the routine production of copper-64.</a> Applied Radiation and Isotopes: Including Data, Instrumentation and Methods for Use in Agriculture, Industry and Medicine. 70 (8), 1803-1806 (2012).</li><li>Ohya, T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Efficient+preparation+of+high-quality+64Cu+for+routine+use.">Efficient preparation of high-quality 64Cu for routine use.</a> Nuclear Medicine and Biology. 43 (11), 685-691 (2016).</li><li>Koole, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=EANM+guidelines+for+PET-CT+and+PET-MR+routine+quality+control.">EANM guidelines for PET-CT and PET-MR routine quality control.</a> Zeitschrift f&#252;r Medizinische Physik. , (2022).</li><li>Sandahl, T. D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+pathophysiology+of+Wilson's+disease+visualized:+A+human+64Cu+PET+study.">The pathophysiology of Wilson's disease visualized: A human 64Cu PET study.</a> Hepatology. 76 (6), 1461-1470 (2022).</li><li>Munk, D. E., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+oral+zinc+regimens+on+human+hepatic+copper+content:+a+randomized+intervention+study.">Effect of oral zinc regimens on human hepatic copper content: a randomized intervention study.</a> Scientific Reports. 12 (1), 14714 (2022).</li><li>Kj&#230;rgaard, K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intravenous+and+oral+copper+kinetics,+biodistribution+and+dosimetry+in+healthy+humans+studied+by+64Cu]copper+PET/CT.">Intravenous and oral copper kinetics, biodistribution and dosimetry in healthy humans studied by 64Cu]copper PET/CT.</a> EJNMMI Radiopharmacy and Chemistry. 5 (1), 15 (2020).</li><li>Brewer, G. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Zinc+acetate+for+the+treatment+of+Wilson's+disease.">Zinc acetate for the treatment of Wilson's disease.</a> Expert Opinion on Pharmacotherapy. 2 (9), 1473-1477 (2001).</li><li>Bush, J. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Studies+on+copper+metabolism.+XVI.+Radioactive+copper+studies+in+normal+subjects+and+in+patients+with+hepatolenticular+degeneration.">Studies on copper metabolism. XVI. Radioactive copper studies in normal subjects and in patients with hepatolenticular degeneration.</a> Journal of Clinical Investigation. 34 (12), 1766-1778 (1955).</li><li>Murillo, O., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=High+value+of+64Cu+as+a+tool+to+evaluate+the+restoration+of+physiological+copper+excretion+after+gene+therapy+in+Wilson's+disease.">High value of 64Cu as a tool to evaluate the restoration of physiological copper excretion after gene therapy in Wilson's disease.</a> Molecular Therapy - Methods &amp; Clinical Development. 26, 98-106 (2022).</li><li>Squitti, R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Copper+dyshomeostasis+in+Wilson+disease+and+Alzheimer's+disease+as+shown+by+serum+and+urine+copper+indicators.">Copper dyshomeostasis in Wilson disease and Alzheimer's disease as shown by serum and urine copper indicators.</a> Journal of Trace Elements in Medicine and Biology. 45, 181-188 (2018).</li></ol>

The authors have no conflicts of interest.

The method is like any other PET method, but the long half-life of 12.7 h offers the opportunity to investigate long-term copper fluxes (we have good results from up to 68 h after IV tracer injection). All steps in the protocol must be handled by personnel familiar with PET, although they are no more critical than any other PET examination.
Troubleshooting 
Because we often use 64Cu for long-term investigations, the PET signal will be noisier than usual. This i...

Copper is a vital catalytic cofactor that drives multiple important biochemical processes essential for life, and defects in copper homeostasis are directly responsible for human diseases. Mutations in the ATP7A or ATP7B genes, encoding copper-transporting ATPases, cause Menke&#39;s and Wilson diseases, respectively. Menke&#39;s disease (ATP7A) is a rare lethal disorder of intestinal copper hyperaccumulation with severe copper deficiency in peripheral tissues and deficits in copper-dependent enzymes1. Wilson disease (WD) (ATP7B) is a rare disease characterized by the inability to excrete excess copper to bile, resulting in copper overload and subsequent organ damage, most severely affecting the liver and brain2.
Studies on copper metabolism have utilized radiolabeled copper (usually 64-copper [64Cu] or 67-copper) for decades, and these studies have proven invaluable for our understanding of mammalian copper metabolism, including absorption site and excretion pathways3,4,5,6. Previously, gamma counters were used to detect the radioactive signal with a limited anatomical resolution, but recently, 64Cu positron emission tomography (PET) combined with computed tomography (CT) or magnetic resonance imaging (MRI) has been introduced in both human and animal studies. Today, PET scanners have such a high sensitivity that it is possible to track 64Cu for up to 70 h after injection. The long half-life of 12.7 h for 64Cu allows for the long-term assessment of copper fluxes. This improvement in resolution has just recently entered the field of copper studies, and studies on normal and pathological copper metabolism, as well as studies evaluating the impact of specific treatments, are starting to emerge. Additionally, the introduction of whole-body PET scanners with an extended field-of-view will further enhance the sensitivity of these examinations.
This methodological paper aims to enable clinicians and scientists to add 64Cu PET CT/MRI to the existing repertoire of tools as a robust and easy-to-use method for assessing copper metabolism in a manner comparable between nuclear medicine departments. The production of 64Cu copper can be carried out using different methods and is usually performed at special facilities. Among the nuclear reactions, the 64Ni (p, n) 64Cu method is widely used, since a high production yield of 64Cu can be obtained with low energy protons in this route7,8. A detailed description of the production methods is out of the scope of this work, and availability will differ by country and region.
In this article, we first describe the preparation of the necessary radiochemistry and the tracer. Then, the principles for preparing the PET/CT or PET/MRI scanners are demonstrated.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>0.22 micrometer sterilizing filter</td>
			<td>Merck Life Science</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Cannula 21 G 50 mm</td>
			<td>BD Microlance</td>
			<td>301155</td>
			<td></td>
		</tr>
		<tr>
			<td>Cannula 25 G 16 mm</td>
			<td>BD Microlance</td>
			<td>300600</td>
			<td></td>
		</tr>
		<tr>
			<td>Dose calibrator</td>
			<td>Capintec CRC-PC calibrator</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>PET/CT scanner</td>
			<td>Siemens: Biograph</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>PET/MR scanner</td>
			<td>GE Signa</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>PMOD version 4.0</td>
			<td>PMOD Technologies LLC</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Saline solution 0.9% NaCl</td>
			<td>Fresenius Kabi</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Sodium acetate trihydrate BioUltra</td>
			<td>Sigma Aldrich</td>
			<td>71188</td>
			<td></td>
		</tr>
		<tr>
			<td>Solid 64CuCl2</td>
			<td>Danish Technical University Ris&#248;</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Sterile water</td>
			<td>Fresenius Kabi</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Venflon 22 G 25 mm</td>
			<td>BD Venflon Pro Safety</td>
			<td>393280</td>
			<td></td>
		</tr>
	</tbody>
</table>

positron emission tomography using 64-copper as a tracer for the study of copper-related disorders

Copper is an essential trace element, functioning in catalysis and signaling in biological systems. Radiolabeled copper has been used for decades in studying basic human and animal copper metabolism and copper-related disorders, such as Wilson disease (WD) and Menke&#39;s disease. A recent addition to this toolkit is 64-copper (64Cu) positron emission tomography (PET), combining the accurate anatomical imaging of modern computed tomography (CT) or magnetic resonance imaging (MRI) scanners with the biodistribution of the 64Cu PET tracer signal. This allows the in vivo tracking of copper fluxes and kinetics, thereby directly visualizing human and animal copper organ traffic and metabolism. Consequently, 64Cu PET is well-suited for evaluating clinical and preclinical treatment effects and has already demonstrated the ability to diagnose WD accurately. Furthermore, 64Cu PET/CT studies have proven valuable in other scientific areas like cancer and stroke research. The present article shows how to perform 64Cu PET/CT or PET/MR in humans. Procedures for 64Cu handling, patient preparation, and scanner setup are demonstrated here.

Dose calculation 
Based on dosimetry calculations, the effective radioactivity dose for IV administration is 62 &#177; 5 &#181;Sv/MBq tracer10. Thus, a 50 MBq dose is recommended depending on the time frame. Up to 75-80 MBq is applicable for longer examinations and provides good-quality images without exceeding an ethically approved dose. The effective dose for oral administration is 113 &#177; 1 &#181;Sv/MBq tracer, due to intestinal accumulation of the tracer. Thus, a lower...

Watch this Scientific Journal Video about Positron Emission Tomography Using 64-Copper as a Tracer for the Study of Copper-Related Disorders at JoVE.com

Positron Emission Tomography Using 64-Copper as a Tracer for the Study of Copper-Related Disorders

The present protocol describes how to perform 64Cu PET/CT and PET/MRI imaging in humans to study copper-related disorders, such as Wilson disease, and the treatment effect on copper metabolism.

Copper is an essential trace element, functioning in catalysis and signaling in biological systems. Radiolabeled copper has been used for decades in ...

This method has already proven to be significant in the study of Wilson disease because we can visualize copper distribution in patients. The benefit of copper PET imaging lies in the ability to monitor its bio distribution within the body, facilitating non-invasive visualization and quantification of the copper concentration in various organs. This technique can be used in the diagnosis of Wilson disease and in research settings for testing treatment effects.In the future, the technique may also be used in clinical settings for treatment effect monitoring, and perhaps in other diseases with disturbed copper metabolism. To begin, wear plastic gloves. Using long tweezers, thoroughly disinfect the rubber membrane of the tracer containing glass bottle inside the lead container with a disinfection swab.Insert a short canula into the membrane using tweezers to prevent spillover from the vacuum inside the bottle. With tweezers, insert a longer canula into the membrane to draw the tracer solution. Insert an appropriately-sized plastic syringe into the long canula and draw the calculated volume of the tracer.Using tweezers, attach a canula with a cap to close the syringe and carefully transfer the syringe to the dose calibrator to measure the radioactivity. Store it in a lead container until use. Measure the radioactivity in the syringe using the dose calibrator.Note the time and activity on the worksheet. Wearing plastic gloves, carefully remove the cap and canula from the syringe using sterile tweezers. Connect the syringe to the patient's IV access.Note the time on the worksheet. And inject the tracer in one steady movement. After injection, remove the syringe from the IV access.Replace the cap and canula on the syringe and keep it that way till the remaining radioactivity is measured. Flush the IV access with saline solution to clear any remaining tracer. Note the time again.Then measure the remaining radioactivity present in the syringe and note it on the worksheet. For oral administration, which is another way of administering the tracer, pour 100 milliliters of cordial into a disposable soft plastic cup. Using tweezers, remove the cap and canula from the syringe.Then while wearing plastic gloves, carefully inject the tracer into the cup without spilling. Draw up a small amount of the cordial into the syringe and inject it back into the cup to ensure proper mixing. Place a plastic straw in the cup and hand over the cup to the participant.Note the time on the worksheet. And allow the participant to drink the mixture. Place the empty cup, straw, and syringe into a disposable plastic bag and place them in the lead container if possible.Note the time and measure the leftover radioactivity in the disposable bag with the syringe, plastic cup, and straw. Place the participant in a supine position in the scanner. Adjust the position of the scanner before beginning the scan.Conduct an overview, CT, or MR scan to plan the specific region to be examined during the PET scan. In the PET scan protocol, note the time of the draw, injection, leftover measurement, and radioactivity levels at the draw and leftover measurement. Download the scan data as DICOM files and transfer them to an analysis program, like PMOD.Adjust the tone settings of scans to differentiate the anatomical structures. Working in the horizontal plane, localize the liver and the big structures. Place appropriate volume of interest or multiple VOIs in the right lobe of the liver at different horizontal planes to obtain the most accurate standard uptake value measurements, considering that the value may vary by 5%within the right liver lobe.Calculate the mean standard uptake value of these VOIs. In a study, comparing healthy subjects and patients with Wilson disease, the distribution of copper was monitored at six and 20 hours post-injection. In patients, copper was accumulating in the liver.Whereas in healthy subjects, the copper is visible in gut segments with higher signals, indicating biliary copper excretion. 64 Copper scans were conducted utilizing an orally-administered tracer in two separate individuals with Wilson disease with and without zinc treatment. Zinc treatment effectively reduces copper absorption in the intestinal tract and subsequently lowers copper accumulation in the liver.In a healthy individual, four weeks of zinc treatment reduced copper content in the liver to around 50%of the pre-treatment content in the group. Remember to look for big structures in the liver when placing the VOIs. And remember to adjust the VOI size to the organ size to avoid spillover from other organs.The procedure can be cobbled with arterial blood sampling in order to perform kinetic analysis. And in addition to blood, copper-64 activity may also be measured in urine and feces.

positron-emission-tomography-using-64-copper-as-tracer-for-study

Research

JoVE Journal

Medicine

1.1K 조회수.  Aarhus University Hospital. 이 방법은 환자의 구리 분포를 시각화 할 수 있기 때문에 윌슨 병 연구에서 이미 중요한 것으로 입증되었습니다. 구리 PET 이미징의 이점은 신체 내 생체 분포를 모니터링하여 다양한 장기에서 구리 농도의 비침습적 시각화 및 정량화를 용이하게 하는 능력에 있습니다. 이 기술은 윌슨병의 진단과 치료 효과를 테스트하기 위한 연구 환경에서 사용할 수 있습니다.앞으로 ?...