Metabolism of 3′-deoxy-3′-[F-18]fluorothymidine in proliferating A549 cells

Validations for positron emission tomography

John R. Grierson, Jeffery L. Schwartz, Mark Muzi, Robert Jordan, Kenneth Krohn

Research output: Contribution to journalArticle

87 Citations (Scopus)

Abstract

3′-Deoxy-3′-[F-18]fluorothymidine (FLT) is under clinical evaluation as a metabolic probe for imaging cell proliferation in vivo using positron emission tomography (PET). As part of our validation effort, we followed the short-term metabolism of FLT in exponentially growing tumor cells to demonstrate the enzyme activities within the DNA salvage pathway that influence retention of radioactivity. In A549 cells, thymidine kinase-1 (TK1) activity produced FLTMP, which dominated the labeled nucleotide pool. Subsequent nucleotide phosphorylations by thymidylate kinase (TMPK) and nucleotide diphosphate kinase (NDPK) led to FLTTP. After 1h, the cellular metabolic pool contained ∼30% FLTTP. A putative deoxynucleotidase (dNT), which degrades FLTMP to FLT, provided the primary mechanism for tracer efflux from cells. In contrast, FLTTP was resistant to degradation and highly retained. The uptake and retention characteristics of FLT were also compared to those of thymidine, FMAU (2′-arabino-fluoro-TdR) and FIAU (2′-arabino-fluoro-5-iodo- 2′-dexoyuridine). Despite the fact that FLT lacks the 3′-hydroxy necessary for its incorporation into DNA it out performed both FMAU and FIAU in terms of uptake and retention.

Original languageEnglish (US)
Pages (from-to)829-837
Number of pages9
JournalNuclear Medicine and Biology
Volume31
Issue number7
DOIs
StatePublished - Oct 2004
Externally publishedYes

Fingerprint

Positron-Emission Tomography
Nucleotides
dTMP kinase
Diphosphates
DNA
Thymidine
Radioactivity
Phosphotransferases
Phosphorylation
Cell Proliferation
Enzymes
A549 Cells
Neoplasms
Clevudine
fialuridine
deoxynucleotidase
thymidine kinase 1

Keywords

  • DNA salvage pathway
  • FLT
  • Fluorothymidine
  • Nucleoside metabolism
  • PET

ASJC Scopus subject areas

  • Cancer Research
  • Molecular Medicine
  • Radiology Nuclear Medicine and imaging

Cite this

Metabolism of 3′-deoxy-3′-[F-18]fluorothymidine in proliferating A549 cells : Validations for positron emission tomography. / Grierson, John R.; Schwartz, Jeffery L.; Muzi, Mark; Jordan, Robert; Krohn, Kenneth.

In: Nuclear Medicine and Biology, Vol. 31, No. 7, 10.2004, p. 829-837.

Research output: Contribution to journalArticle

Grierson, John R. ; Schwartz, Jeffery L. ; Muzi, Mark ; Jordan, Robert ; Krohn, Kenneth. / Metabolism of 3′-deoxy-3′-[F-18]fluorothymidine in proliferating A549 cells : Validations for positron emission tomography. In: Nuclear Medicine and Biology. 2004 ; Vol. 31, No. 7. pp. 829-837.
@article{5f3855f3d908424cb5ea7d0e2e6e7f5a,
title = "Metabolism of 3′-deoxy-3′-[F-18]fluorothymidine in proliferating A549 cells: Validations for positron emission tomography",
abstract = "3′-Deoxy-3′-[F-18]fluorothymidine (FLT) is under clinical evaluation as a metabolic probe for imaging cell proliferation in vivo using positron emission tomography (PET). As part of our validation effort, we followed the short-term metabolism of FLT in exponentially growing tumor cells to demonstrate the enzyme activities within the DNA salvage pathway that influence retention of radioactivity. In A549 cells, thymidine kinase-1 (TK1) activity produced FLTMP, which dominated the labeled nucleotide pool. Subsequent nucleotide phosphorylations by thymidylate kinase (TMPK) and nucleotide diphosphate kinase (NDPK) led to FLTTP. After 1h, the cellular metabolic pool contained ∼30{\%} FLTTP. A putative deoxynucleotidase (dNT), which degrades FLTMP to FLT, provided the primary mechanism for tracer efflux from cells. In contrast, FLTTP was resistant to degradation and highly retained. The uptake and retention characteristics of FLT were also compared to those of thymidine, FMAU (2′-arabino-fluoro-TdR) and FIAU (2′-arabino-fluoro-5-iodo- 2′-dexoyuridine). Despite the fact that FLT lacks the 3′-hydroxy necessary for its incorporation into DNA it out performed both FMAU and FIAU in terms of uptake and retention.",
keywords = "DNA salvage pathway, FLT, Fluorothymidine, Nucleoside metabolism, PET",
author = "Grierson, {John R.} and Schwartz, {Jeffery L.} and Mark Muzi and Robert Jordan and Kenneth Krohn",
year = "2004",
month = "10",
doi = "10.1016/j.nucmedbio.2004.06.004",
language = "English (US)",
volume = "31",
pages = "829--837",
journal = "International journal of radiation applications and instrumentation. Part B, Nuclear medicine and biology",
issn = "0969-8051",
publisher = "Elsevier Inc.",
number = "7",

}

TY - JOUR

T1 - Metabolism of 3′-deoxy-3′-[F-18]fluorothymidine in proliferating A549 cells

T2 - Validations for positron emission tomography

AU - Grierson, John R.

AU - Schwartz, Jeffery L.

AU - Muzi, Mark

AU - Jordan, Robert

AU - Krohn, Kenneth

PY - 2004/10

Y1 - 2004/10

N2 - 3′-Deoxy-3′-[F-18]fluorothymidine (FLT) is under clinical evaluation as a metabolic probe for imaging cell proliferation in vivo using positron emission tomography (PET). As part of our validation effort, we followed the short-term metabolism of FLT in exponentially growing tumor cells to demonstrate the enzyme activities within the DNA salvage pathway that influence retention of radioactivity. In A549 cells, thymidine kinase-1 (TK1) activity produced FLTMP, which dominated the labeled nucleotide pool. Subsequent nucleotide phosphorylations by thymidylate kinase (TMPK) and nucleotide diphosphate kinase (NDPK) led to FLTTP. After 1h, the cellular metabolic pool contained ∼30% FLTTP. A putative deoxynucleotidase (dNT), which degrades FLTMP to FLT, provided the primary mechanism for tracer efflux from cells. In contrast, FLTTP was resistant to degradation and highly retained. The uptake and retention characteristics of FLT were also compared to those of thymidine, FMAU (2′-arabino-fluoro-TdR) and FIAU (2′-arabino-fluoro-5-iodo- 2′-dexoyuridine). Despite the fact that FLT lacks the 3′-hydroxy necessary for its incorporation into DNA it out performed both FMAU and FIAU in terms of uptake and retention.

AB - 3′-Deoxy-3′-[F-18]fluorothymidine (FLT) is under clinical evaluation as a metabolic probe for imaging cell proliferation in vivo using positron emission tomography (PET). As part of our validation effort, we followed the short-term metabolism of FLT in exponentially growing tumor cells to demonstrate the enzyme activities within the DNA salvage pathway that influence retention of radioactivity. In A549 cells, thymidine kinase-1 (TK1) activity produced FLTMP, which dominated the labeled nucleotide pool. Subsequent nucleotide phosphorylations by thymidylate kinase (TMPK) and nucleotide diphosphate kinase (NDPK) led to FLTTP. After 1h, the cellular metabolic pool contained ∼30% FLTTP. A putative deoxynucleotidase (dNT), which degrades FLTMP to FLT, provided the primary mechanism for tracer efflux from cells. In contrast, FLTTP was resistant to degradation and highly retained. The uptake and retention characteristics of FLT were also compared to those of thymidine, FMAU (2′-arabino-fluoro-TdR) and FIAU (2′-arabino-fluoro-5-iodo- 2′-dexoyuridine). Despite the fact that FLT lacks the 3′-hydroxy necessary for its incorporation into DNA it out performed both FMAU and FIAU in terms of uptake and retention.

KW - DNA salvage pathway

KW - FLT

KW - Fluorothymidine

KW - Nucleoside metabolism

KW - PET

UR - http://www.scopus.com/inward/record.url?scp=4644361597&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=4644361597&partnerID=8YFLogxK

U2 - 10.1016/j.nucmedbio.2004.06.004

DO - 10.1016/j.nucmedbio.2004.06.004

M3 - Article

VL - 31

SP - 829

EP - 837

JO - International journal of radiation applications and instrumentation. Part B, Nuclear medicine and biology

JF - International journal of radiation applications and instrumentation. Part B, Nuclear medicine and biology

SN - 0969-8051

IS - 7

ER -