Human islet cell MORF/cMORF pretargeting in a xenogeneic murine transplant model

Guozheng Liu, Shuping Dou, Dengfeng Cheng, Jean Leif, Mary Rusckowski, Philip Streeter, Leonard D. Shultz, Donald J. Hnatowich, Dale L. Greiner

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

noninvasive measurement of human islet cell mass in pancreas or following islet transplantation by nuclear imaging has yet to be achieved. It has been shown using mouse tumor models that pretargeting imaging strategies are sensitive and can greatly increase target to nontarget signal ratios. The objective now is to demonstrate the specific pretargeting of human islet cells in mice. Our pretargeting strategy uses an anti-human islet cell antibody HPi1, conjugated to a phosphorodiamidate morpholino oligomer (MORF) that binds specifically to a 99mTc labeled complementary MORF (cMORF). Sensitivity and specificity of the pretargeting were first validated in culture using a human beta cell line (betalox5) and a negative control human cell line (HEK293). Pretargeting was then used to target and visualize these two cell lines and human islets transplanted subcutaneously in NOD-scid IL2rγnull mice. In culture, 99mTc accumulation on the betalox5 cells pretargeted by MORF-HPi1 was 100-fold higher than on untreated betalox5 cells or following treatment with native HPi1 and much higher than on the MORF-HPi1 pretargeted control HEK293 cells. Small animal imaging readily localized the transplanted betalox5 cells and human islets, but not the HEK293 cells. Ex vivo counting demonstrated 3-fold higher 99mTc accumulation in the transplanted betalox5 cells and human islets than in the control HEK293 cells. The target accumulation was also shown to increase linearly with increased numbers of the implanted betalox5 cells. These results demonstrate specific binding of radioactivity and successful imaging of human betalox5 cells and human islets transplanted in mice. Thus MORF/cMORF pretargeting may be useful to measure noninvasively human islet cell mass within the pancreas or following islet transplantation.

Original languageEnglish (US)
Pages (from-to)767-773
Number of pages7
JournalMolecular Pharmaceutics
Volume8
Issue number3
DOIs
StatePublished - Jun 6 2011

Fingerprint

Islets of Langerhans
Transplants
HEK293 Cells
Islets of Langerhans Transplantation
Cell Line
Pancreas
Morpholinos
Radioactivity
Sensitivity and Specificity

ASJC Scopus subject areas

  • Pharmaceutical Science
  • Molecular Medicine
  • Drug Discovery

Cite this

Liu, G., Dou, S., Cheng, D., Leif, J., Rusckowski, M., Streeter, P., ... Greiner, D. L. (2011). Human islet cell MORF/cMORF pretargeting in a xenogeneic murine transplant model. Molecular Pharmaceutics, 8(3), 767-773. https://doi.org/10.1021/mp100382m

Human islet cell MORF/cMORF pretargeting in a xenogeneic murine transplant model. / Liu, Guozheng; Dou, Shuping; Cheng, Dengfeng; Leif, Jean; Rusckowski, Mary; Streeter, Philip; Shultz, Leonard D.; Hnatowich, Donald J.; Greiner, Dale L.

In: Molecular Pharmaceutics, Vol. 8, No. 3, 06.06.2011, p. 767-773.

Research output: Contribution to journalArticle

Liu, G, Dou, S, Cheng, D, Leif, J, Rusckowski, M, Streeter, P, Shultz, LD, Hnatowich, DJ & Greiner, DL 2011, 'Human islet cell MORF/cMORF pretargeting in a xenogeneic murine transplant model', Molecular Pharmaceutics, vol. 8, no. 3, pp. 767-773. https://doi.org/10.1021/mp100382m
Liu G, Dou S, Cheng D, Leif J, Rusckowski M, Streeter P et al. Human islet cell MORF/cMORF pretargeting in a xenogeneic murine transplant model. Molecular Pharmaceutics. 2011 Jun 6;8(3):767-773. https://doi.org/10.1021/mp100382m
Liu, Guozheng ; Dou, Shuping ; Cheng, Dengfeng ; Leif, Jean ; Rusckowski, Mary ; Streeter, Philip ; Shultz, Leonard D. ; Hnatowich, Donald J. ; Greiner, Dale L. / Human islet cell MORF/cMORF pretargeting in a xenogeneic murine transplant model. In: Molecular Pharmaceutics. 2011 ; Vol. 8, No. 3. pp. 767-773.
@article{833554f6603041bdb837c22418ce7b64,
title = "Human islet cell MORF/cMORF pretargeting in a xenogeneic murine transplant model",
abstract = "noninvasive measurement of human islet cell mass in pancreas or following islet transplantation by nuclear imaging has yet to be achieved. It has been shown using mouse tumor models that pretargeting imaging strategies are sensitive and can greatly increase target to nontarget signal ratios. The objective now is to demonstrate the specific pretargeting of human islet cells in mice. Our pretargeting strategy uses an anti-human islet cell antibody HPi1, conjugated to a phosphorodiamidate morpholino oligomer (MORF) that binds specifically to a 99mTc labeled complementary MORF (cMORF). Sensitivity and specificity of the pretargeting were first validated in culture using a human beta cell line (betalox5) and a negative control human cell line (HEK293). Pretargeting was then used to target and visualize these two cell lines and human islets transplanted subcutaneously in NOD-scid IL2rγnull mice. In culture, 99mTc accumulation on the betalox5 cells pretargeted by MORF-HPi1 was 100-fold higher than on untreated betalox5 cells or following treatment with native HPi1 and much higher than on the MORF-HPi1 pretargeted control HEK293 cells. Small animal imaging readily localized the transplanted betalox5 cells and human islets, but not the HEK293 cells. Ex vivo counting demonstrated 3-fold higher 99mTc accumulation in the transplanted betalox5 cells and human islets than in the control HEK293 cells. The target accumulation was also shown to increase linearly with increased numbers of the implanted betalox5 cells. These results demonstrate specific binding of radioactivity and successful imaging of human betalox5 cells and human islets transplanted in mice. Thus MORF/cMORF pretargeting may be useful to measure noninvasively human islet cell mass within the pancreas or following islet transplantation.",
author = "Guozheng Liu and Shuping Dou and Dengfeng Cheng and Jean Leif and Mary Rusckowski and Philip Streeter and Shultz, {Leonard D.} and Hnatowich, {Donald J.} and Greiner, {Dale L.}",
year = "2011",
month = "6",
day = "6",
doi = "10.1021/mp100382m",
language = "English (US)",
volume = "8",
pages = "767--773",
journal = "Molecular Pharmaceutics",
issn = "1543-8384",
publisher = "American Chemical Society",
number = "3",

}

TY - JOUR

T1 - Human islet cell MORF/cMORF pretargeting in a xenogeneic murine transplant model

AU - Liu, Guozheng

AU - Dou, Shuping

AU - Cheng, Dengfeng

AU - Leif, Jean

AU - Rusckowski, Mary

AU - Streeter, Philip

AU - Shultz, Leonard D.

AU - Hnatowich, Donald J.

AU - Greiner, Dale L.

PY - 2011/6/6

Y1 - 2011/6/6

N2 - noninvasive measurement of human islet cell mass in pancreas or following islet transplantation by nuclear imaging has yet to be achieved. It has been shown using mouse tumor models that pretargeting imaging strategies are sensitive and can greatly increase target to nontarget signal ratios. The objective now is to demonstrate the specific pretargeting of human islet cells in mice. Our pretargeting strategy uses an anti-human islet cell antibody HPi1, conjugated to a phosphorodiamidate morpholino oligomer (MORF) that binds specifically to a 99mTc labeled complementary MORF (cMORF). Sensitivity and specificity of the pretargeting were first validated in culture using a human beta cell line (betalox5) and a negative control human cell line (HEK293). Pretargeting was then used to target and visualize these two cell lines and human islets transplanted subcutaneously in NOD-scid IL2rγnull mice. In culture, 99mTc accumulation on the betalox5 cells pretargeted by MORF-HPi1 was 100-fold higher than on untreated betalox5 cells or following treatment with native HPi1 and much higher than on the MORF-HPi1 pretargeted control HEK293 cells. Small animal imaging readily localized the transplanted betalox5 cells and human islets, but not the HEK293 cells. Ex vivo counting demonstrated 3-fold higher 99mTc accumulation in the transplanted betalox5 cells and human islets than in the control HEK293 cells. The target accumulation was also shown to increase linearly with increased numbers of the implanted betalox5 cells. These results demonstrate specific binding of radioactivity and successful imaging of human betalox5 cells and human islets transplanted in mice. Thus MORF/cMORF pretargeting may be useful to measure noninvasively human islet cell mass within the pancreas or following islet transplantation.

AB - noninvasive measurement of human islet cell mass in pancreas or following islet transplantation by nuclear imaging has yet to be achieved. It has been shown using mouse tumor models that pretargeting imaging strategies are sensitive and can greatly increase target to nontarget signal ratios. The objective now is to demonstrate the specific pretargeting of human islet cells in mice. Our pretargeting strategy uses an anti-human islet cell antibody HPi1, conjugated to a phosphorodiamidate morpholino oligomer (MORF) that binds specifically to a 99mTc labeled complementary MORF (cMORF). Sensitivity and specificity of the pretargeting were first validated in culture using a human beta cell line (betalox5) and a negative control human cell line (HEK293). Pretargeting was then used to target and visualize these two cell lines and human islets transplanted subcutaneously in NOD-scid IL2rγnull mice. In culture, 99mTc accumulation on the betalox5 cells pretargeted by MORF-HPi1 was 100-fold higher than on untreated betalox5 cells or following treatment with native HPi1 and much higher than on the MORF-HPi1 pretargeted control HEK293 cells. Small animal imaging readily localized the transplanted betalox5 cells and human islets, but not the HEK293 cells. Ex vivo counting demonstrated 3-fold higher 99mTc accumulation in the transplanted betalox5 cells and human islets than in the control HEK293 cells. The target accumulation was also shown to increase linearly with increased numbers of the implanted betalox5 cells. These results demonstrate specific binding of radioactivity and successful imaging of human betalox5 cells and human islets transplanted in mice. Thus MORF/cMORF pretargeting may be useful to measure noninvasively human islet cell mass within the pancreas or following islet transplantation.

UR - http://www.scopus.com/inward/record.url?scp=79958297561&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=79958297561&partnerID=8YFLogxK

U2 - 10.1021/mp100382m

DO - 10.1021/mp100382m

M3 - Article

C2 - 21361360

AN - SCOPUS:79958297561

VL - 8

SP - 767

EP - 773

JO - Molecular Pharmaceutics

JF - Molecular Pharmaceutics

SN - 1543-8384

IS - 3

ER -