Evidence of 68 Ga-DOTA-NT-20.3 uptake in pancreatic adenocarcinoma AsPC-1 cell line – In vitro study

Manuela Marenco, Lorenzo Lodola, Marco G. Persico, Vanessa Frangipane, Angelica Facoetti, Carlo Aprile, Marina Hodolič

Research output: Contribution to journalArticle

Abstract

Background: Neurotensin receptors are overexpressed in several cancer types including pancreatic ductal adenocarcinoma. Three NTR subtypes have been cloned: NTR-1, NTR-2 and NTR-3. The most expressed NTR-1 is not present in normal pancreatic tissue and has a low expression in chronic pancreatitis. Objective: Objective of this study was to test in vitro affinity of the new68 Ga labelled neurotensin analogue DOTA-NT-20.3 (fragment 6-13, Ac-Lys(DOTA)-Pro-Arg(N-CH3 )-Arg-Pro-Tyr-Tle-Leu) on the human pancreatic ductal adenocarcinoma cell line AsPC-1. Method: For the preparation of68 Ga-DOTA-NT-20.3,68GaCl3 solution (eluted from68 Ge/68 Ga generator) and 50 µg of precursor (Iason, Graz, Austria) water dissolved were used in an automatic synthesis module. The labeled compound was added to cell culture flask and incubated at 37°C. At various time points after tracer addition up to 80min, cells were recovered, rinsed and counted for radioactivity. Results were expressed as percent binding normalized to 200000 cells and affinity parameters were calculated. Results: Labeling yield was ≥98 %. The molar ratio between labelled and total peptide was about 1/400. AsPC-1 cell line showed rapid uptake of the tracer including surface and internalized binding, tending to a plateau phase 80 min after tracer addition (11%/200.000 cells). The Kd (7.335 pmol) and Bmax (90.52 kBq) value indicated high tracer affinity for AsPC-1cell line especially if compared with the literature data regarding other malignancies (e.g. colonic cancer cell line). Binding sites were 1.09x106 sites per cell. Conclusion: New tracer68 Ga-DOTA-NT-20.3 can be a suitable candidate for the clinical use in patients with pancreatic ductal adenocarcinoma.

Original languageEnglish
Pages (from-to)754-759
Number of pages6
JournalCurrent Pharmaceutical Biotechnology
Volume19
Issue number9
DOIs
Publication statusPublished - Jan 1 2018

Fingerprint

Adenocarcinoma
Cell Line
prolylarginine
Neurotensin Receptors
Neurotensin
Austria
Chronic Pancreatitis
Pancreatic Neoplasms
Colonic Neoplasms
Radioactivity
Cell Culture Techniques
Binding Sites
Peptides
In Vitro Techniques
Water
Neoplasms

Keywords

  • Ga-NT-DOTA-20.3
  • AsPC-1
  • Neurotensin
  • Neurotensin receptors
  • NT
  • NTR-1
  • Pancreatic ductal adenocarcinoma

ASJC Scopus subject areas

  • Biotechnology
  • Pharmaceutical Science

Cite this

Evidence of 68 Ga-DOTA-NT-20.3 uptake in pancreatic adenocarcinoma AsPC-1 cell line – In vitro study. / Marenco, Manuela; Lodola, Lorenzo; Persico, Marco G.; Frangipane, Vanessa; Facoetti, Angelica; Aprile, Carlo; Hodolič, Marina.

In: Current Pharmaceutical Biotechnology, Vol. 19, No. 9, 01.01.2018, p. 754-759.

Research output: Contribution to journalArticle

Marenco, Manuela ; Lodola, Lorenzo ; Persico, Marco G. ; Frangipane, Vanessa ; Facoetti, Angelica ; Aprile, Carlo ; Hodolič, Marina. / Evidence of 68 Ga-DOTA-NT-20.3 uptake in pancreatic adenocarcinoma AsPC-1 cell line – In vitro study. In: Current Pharmaceutical Biotechnology. 2018 ; Vol. 19, No. 9. pp. 754-759.
@article{3f2d7b12e3804343b89947ce98c41cb7,
title = "Evidence of 68 Ga-DOTA-NT-20.3 uptake in pancreatic adenocarcinoma AsPC-1 cell line – In vitro study",
abstract = "Background: Neurotensin receptors are overexpressed in several cancer types including pancreatic ductal adenocarcinoma. Three NTR subtypes have been cloned: NTR-1, NTR-2 and NTR-3. The most expressed NTR-1 is not present in normal pancreatic tissue and has a low expression in chronic pancreatitis. Objective: Objective of this study was to test in vitro affinity of the new68 Ga labelled neurotensin analogue DOTA-NT-20.3 (fragment 6-13, Ac-Lys(DOTA)-Pro-Arg(N-CH3 )-Arg-Pro-Tyr-Tle-Leu) on the human pancreatic ductal adenocarcinoma cell line AsPC-1. Method: For the preparation of68 Ga-DOTA-NT-20.3,68GaCl3 solution (eluted from68 Ge/68 Ga generator) and 50 µg of precursor (Iason, Graz, Austria) water dissolved were used in an automatic synthesis module. The labeled compound was added to cell culture flask and incubated at 37°C. At various time points after tracer addition up to 80min, cells were recovered, rinsed and counted for radioactivity. Results were expressed as percent binding normalized to 200000 cells and affinity parameters were calculated. Results: Labeling yield was ≥98 {\%}. The molar ratio between labelled and total peptide was about 1/400. AsPC-1 cell line showed rapid uptake of the tracer including surface and internalized binding, tending to a plateau phase 80 min after tracer addition (11{\%}/200.000 cells). The Kd (7.335 pmol) and Bmax (90.52 kBq) value indicated high tracer affinity for AsPC-1cell line especially if compared with the literature data regarding other malignancies (e.g. colonic cancer cell line). Binding sites were 1.09x106 sites per cell. Conclusion: New tracer68 Ga-DOTA-NT-20.3 can be a suitable candidate for the clinical use in patients with pancreatic ductal adenocarcinoma.",
keywords = "Ga-NT-DOTA-20.3, AsPC-1, Neurotensin, Neurotensin receptors, NT, NTR-1, Pancreatic ductal adenocarcinoma",
author = "Manuela Marenco and Lorenzo Lodola and Persico, {Marco G.} and Vanessa Frangipane and Angelica Facoetti and Carlo Aprile and Marina Hodolič",
year = "2018",
month = "1",
day = "1",
doi = "10.2174/1389201019666180829152314",
language = "English",
volume = "19",
pages = "754--759",
journal = "Current Pharmaceutical Biotechnology",
issn = "1389-2010",
publisher = "Bentham Science Publishers B.V.",
number = "9",

}

TY - JOUR

T1 - Evidence of 68 Ga-DOTA-NT-20.3 uptake in pancreatic adenocarcinoma AsPC-1 cell line – In vitro study

AU - Marenco, Manuela

AU - Lodola, Lorenzo

AU - Persico, Marco G.

AU - Frangipane, Vanessa

AU - Facoetti, Angelica

AU - Aprile, Carlo

AU - Hodolič, Marina

PY - 2018/1/1

Y1 - 2018/1/1

N2 - Background: Neurotensin receptors are overexpressed in several cancer types including pancreatic ductal adenocarcinoma. Three NTR subtypes have been cloned: NTR-1, NTR-2 and NTR-3. The most expressed NTR-1 is not present in normal pancreatic tissue and has a low expression in chronic pancreatitis. Objective: Objective of this study was to test in vitro affinity of the new68 Ga labelled neurotensin analogue DOTA-NT-20.3 (fragment 6-13, Ac-Lys(DOTA)-Pro-Arg(N-CH3 )-Arg-Pro-Tyr-Tle-Leu) on the human pancreatic ductal adenocarcinoma cell line AsPC-1. Method: For the preparation of68 Ga-DOTA-NT-20.3,68GaCl3 solution (eluted from68 Ge/68 Ga generator) and 50 µg of precursor (Iason, Graz, Austria) water dissolved were used in an automatic synthesis module. The labeled compound was added to cell culture flask and incubated at 37°C. At various time points after tracer addition up to 80min, cells were recovered, rinsed and counted for radioactivity. Results were expressed as percent binding normalized to 200000 cells and affinity parameters were calculated. Results: Labeling yield was ≥98 %. The molar ratio between labelled and total peptide was about 1/400. AsPC-1 cell line showed rapid uptake of the tracer including surface and internalized binding, tending to a plateau phase 80 min after tracer addition (11%/200.000 cells). The Kd (7.335 pmol) and Bmax (90.52 kBq) value indicated high tracer affinity for AsPC-1cell line especially if compared with the literature data regarding other malignancies (e.g. colonic cancer cell line). Binding sites were 1.09x106 sites per cell. Conclusion: New tracer68 Ga-DOTA-NT-20.3 can be a suitable candidate for the clinical use in patients with pancreatic ductal adenocarcinoma.

AB - Background: Neurotensin receptors are overexpressed in several cancer types including pancreatic ductal adenocarcinoma. Three NTR subtypes have been cloned: NTR-1, NTR-2 and NTR-3. The most expressed NTR-1 is not present in normal pancreatic tissue and has a low expression in chronic pancreatitis. Objective: Objective of this study was to test in vitro affinity of the new68 Ga labelled neurotensin analogue DOTA-NT-20.3 (fragment 6-13, Ac-Lys(DOTA)-Pro-Arg(N-CH3 )-Arg-Pro-Tyr-Tle-Leu) on the human pancreatic ductal adenocarcinoma cell line AsPC-1. Method: For the preparation of68 Ga-DOTA-NT-20.3,68GaCl3 solution (eluted from68 Ge/68 Ga generator) and 50 µg of precursor (Iason, Graz, Austria) water dissolved were used in an automatic synthesis module. The labeled compound was added to cell culture flask and incubated at 37°C. At various time points after tracer addition up to 80min, cells were recovered, rinsed and counted for radioactivity. Results were expressed as percent binding normalized to 200000 cells and affinity parameters were calculated. Results: Labeling yield was ≥98 %. The molar ratio between labelled and total peptide was about 1/400. AsPC-1 cell line showed rapid uptake of the tracer including surface and internalized binding, tending to a plateau phase 80 min after tracer addition (11%/200.000 cells). The Kd (7.335 pmol) and Bmax (90.52 kBq) value indicated high tracer affinity for AsPC-1cell line especially if compared with the literature data regarding other malignancies (e.g. colonic cancer cell line). Binding sites were 1.09x106 sites per cell. Conclusion: New tracer68 Ga-DOTA-NT-20.3 can be a suitable candidate for the clinical use in patients with pancreatic ductal adenocarcinoma.

KW - Ga-NT-DOTA-20.3

KW - AsPC-1

KW - Neurotensin

KW - Neurotensin receptors

KW - NT

KW - NTR-1

KW - Pancreatic ductal adenocarcinoma

UR - http://www.scopus.com/inward/record.url?scp=85057527659&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85057527659&partnerID=8YFLogxK

U2 - 10.2174/1389201019666180829152314

DO - 10.2174/1389201019666180829152314

M3 - Article

AN - SCOPUS:85057527659

VL - 19

SP - 754

EP - 759

JO - Current Pharmaceutical Biotechnology

JF - Current Pharmaceutical Biotechnology

SN - 1389-2010

IS - 9

ER -