NMR spectroscopy in β cell engineering and islet transplantation

Klearchos K Papas, Clark K. Colton, John S. Gounarides, Eric S. Roos, Mary Ann C Jarema, Michael J. Shapiro, Leo L. Cheng, Gary W. Cline, Gerald I. Shulman, W. U. Haiyan, Susan Bonner-Weir, Gordon C. Weir

Research output: Contribution to journalArticle

30 Citations (Scopus)

Abstract

Islet transplantation is a promising method for restoring normoglycemia and alleviating the long term complications of diabetes. Widespread application of islet transplantation is hindered by the limited supply of human islets and requires a large increase in the availability of suitable insulin secreting tissue as well as robust quality assessment methodologies that can ensure safety and in vivo efficacy. We explore the application of nuclear magnetic resonance (NMR) spectroscopy in two areas relevant to β cell engineering and islet transplantation: (1) the effect of genetic alterations on glucose metabolism, and (2) quality assessment of islet preparations prior to transplantation. Results obtained utilizing a variety of NMR techniques demonstrate the following: (1) Transfection of Rat1 cells with the c-myc oncogene (which may be involved in cell proliferation and cell cycle regulation) and overexpression of Bcl-2 (which may protect cells from stresses such as hypoxia and exposure to cytokines) introduce a wide array of alterations in cellular biochemistry, including changes in anaerobic and oxidative glucose metabolism, as assessed by 13C and 31P NMR spectroscopy. (2) Overnight incubation of islets and β cells in the bottom of centrifuge tubes filled with medium at room temperature, as is sometimes done in islet transportation, exposes them to severe oxygen limitations that may cause cell damage. Such exposure, leading to reversible or irreversible damage, can be observed with NMR-detectable markers using conventional 13C and 31P NMR spectroscopy of extracts. In addition, markers of irreversible damage (as well as markers of hypoxia) can be detected and quantified without cell extraction using high-resolution magic angle spinning 1H NMR spectroscopy. Finally, acute ischemia in a bed of perfused β cells leads to completely reversible changes that can be followed in real time with 31P NMR spectroscopy.

Original languageEnglish (US)
Pages (from-to)96-119
Number of pages24
JournalAnnals of the New York Academy of Sciences
Volume944
StatePublished - 2001
Externally publishedYes

Fingerprint

Cell engineering
Transplantation (surgical)
Cell Engineering
Islets of Langerhans Transplantation
Cell Transplantation
Nuclear magnetic resonance spectroscopy
Magnetic Resonance Spectroscopy
Metabolism
Cells
Nuclear magnetic resonance
Glucose
Biochemistry
Magic angle spinning
Centrifuges
Cell proliferation
Medical problems
myc Genes
Diabetes Complications
Nuclear Magnetic Resonance
Transplantation

Keywords

  • β cell
  • Apoptosis
  • Assessment
  • Bcl-2
  • c-myc
  • Culture
  • Hypoxia
  • Islet
  • Lactate
  • LDH
  • Lipids
  • NMR
  • Oxygen
  • Phosphocholine
  • Quality
  • Transplantation
  • Transportation

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

Papas, K. K., Colton, C. K., Gounarides, J. S., Roos, E. S., Jarema, M. A. C., Shapiro, M. J., ... Weir, G. C. (2001). NMR spectroscopy in β cell engineering and islet transplantation. Annals of the New York Academy of Sciences, 944, 96-119.

NMR spectroscopy in β cell engineering and islet transplantation. / Papas, Klearchos K; Colton, Clark K.; Gounarides, John S.; Roos, Eric S.; Jarema, Mary Ann C; Shapiro, Michael J.; Cheng, Leo L.; Cline, Gary W.; Shulman, Gerald I.; Haiyan, W. U.; Bonner-Weir, Susan; Weir, Gordon C.

In: Annals of the New York Academy of Sciences, Vol. 944, 2001, p. 96-119.

Research output: Contribution to journalArticle

Papas, KK, Colton, CK, Gounarides, JS, Roos, ES, Jarema, MAC, Shapiro, MJ, Cheng, LL, Cline, GW, Shulman, GI, Haiyan, WU, Bonner-Weir, S & Weir, GC 2001, 'NMR spectroscopy in β cell engineering and islet transplantation', Annals of the New York Academy of Sciences, vol. 944, pp. 96-119.
Papas KK, Colton CK, Gounarides JS, Roos ES, Jarema MAC, Shapiro MJ et al. NMR spectroscopy in β cell engineering and islet transplantation. Annals of the New York Academy of Sciences. 2001;944:96-119.
Papas, Klearchos K ; Colton, Clark K. ; Gounarides, John S. ; Roos, Eric S. ; Jarema, Mary Ann C ; Shapiro, Michael J. ; Cheng, Leo L. ; Cline, Gary W. ; Shulman, Gerald I. ; Haiyan, W. U. ; Bonner-Weir, Susan ; Weir, Gordon C. / NMR spectroscopy in β cell engineering and islet transplantation. In: Annals of the New York Academy of Sciences. 2001 ; Vol. 944. pp. 96-119.
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AU - Shapiro, Michael J.

AU - Cheng, Leo L.

AU - Cline, Gary W.

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