Glyoxylate reductase activity in blood mononuclear cells and the diagnosis of primary hyperoxaluria type 2

John Knight, Ross P. Holmes, Dawn S. Milliner, Carla G. Monico, Scott D. Cramer

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

Background. Primary hyperoxaluria type 2 (PH2) is a rare monogenic disorder characterized by an elevated urinary excretion of oxalate. Increased oxalate excretion in PH2 patients can cause nephrolithiasis and nephrocalcinosis, and can, in some cases, result in renal failure and systemic oxalate deposition. The disease is due to a deficiency of glyoxylate reductase/hydroxypyruvate reductase (GRHPR) activity. A definitive diagnosis of PH2 is currently made by the analysis of GR activity in a liver biopsy. GRHPR is expressed in virtually every tissue in the body, suggesting that utilization of more readily available cells could be used to determine GRHPR deficiency. In this study, we have evaluated the potential of determining GR and d -glycerate dehydrogenase (DGDH) activity in blood mononuclear cells (BMC) as a diagnostic indicator of PH2. Methods. Blood samples were obtained from 10 male and 10 female normal subjects, median age 31, range 21-63, at the Wake Forest University Medical Center and from primary hyperoxaluria patients at the Mayo Clinic. The BMC were isolated and GR and DGDH activities measured in cell lysates. Results. An assay of 20 normal individuals indicated that BMC contained a DGDH and GR activity of 0.97±0.20 (range 0.62-1.45), and 10.6±3.3 (range 8.3-16.6) nmol/min/mg protein, respectively. The intra-assay coefficient of variation for DGDH and GR activity was 8.2 and 11.5%, respectively. The BMC lysates from al adult subjects and patients with PH1 showed similar GR and DGDH activities. This was confirmed by the presence of immunoreactive GRHPR protein by western blot analysis. In contrast, PH2 BMC lysates did not exhibit DGDH or GR activity, and showed no immunoreactive GRHPR by western blot analysis. Conclusion. These results suggest that the assay of DGDH or GR activity in BMC could be used as a minimally invasive diagnostic test for PH2.

Original languageEnglish (US)
Pages (from-to)2292-2295
Number of pages4
JournalNephrology Dialysis Transplantation
Volume21
Issue number8
DOIs
StatePublished - Aug 2006

Fingerprint

glyoxylate reductase
Glycerate dehydrogenase
Blood Cells
Hydroxypyruvate Reductase
Oxalates
Western Blotting
Primary Hyperoxaluria
Nephrocalcinosis
Nephrolithiasis
Primary hyperoxaluria type 2
Routine Diagnostic Tests

Keywords

  • Blood mononuclear cells
  • D-glycerate dehydrogenase
  • Glyoxylate reductase
  • Primary hyperoxaluria type 2

ASJC Scopus subject areas

  • Nephrology
  • Transplantation

Cite this

Glyoxylate reductase activity in blood mononuclear cells and the diagnosis of primary hyperoxaluria type 2. / Knight, John; Holmes, Ross P.; Milliner, Dawn S.; Monico, Carla G.; Cramer, Scott D.

In: Nephrology Dialysis Transplantation, Vol. 21, No. 8, 08.2006, p. 2292-2295.

Research output: Contribution to journalArticle

Knight, John ; Holmes, Ross P. ; Milliner, Dawn S. ; Monico, Carla G. ; Cramer, Scott D. / Glyoxylate reductase activity in blood mononuclear cells and the diagnosis of primary hyperoxaluria type 2. In: Nephrology Dialysis Transplantation. 2006 ; Vol. 21, No. 8. pp. 2292-2295.
@article{f93893ff5f494b5ebec5f6326bdd133a,
title = "Glyoxylate reductase activity in blood mononuclear cells and the diagnosis of primary hyperoxaluria type 2",
abstract = "Background. Primary hyperoxaluria type 2 (PH2) is a rare monogenic disorder characterized by an elevated urinary excretion of oxalate. Increased oxalate excretion in PH2 patients can cause nephrolithiasis and nephrocalcinosis, and can, in some cases, result in renal failure and systemic oxalate deposition. The disease is due to a deficiency of glyoxylate reductase/hydroxypyruvate reductase (GRHPR) activity. A definitive diagnosis of PH2 is currently made by the analysis of GR activity in a liver biopsy. GRHPR is expressed in virtually every tissue in the body, suggesting that utilization of more readily available cells could be used to determine GRHPR deficiency. In this study, we have evaluated the potential of determining GR and d -glycerate dehydrogenase (DGDH) activity in blood mononuclear cells (BMC) as a diagnostic indicator of PH2. Methods. Blood samples were obtained from 10 male and 10 female normal subjects, median age 31, range 21-63, at the Wake Forest University Medical Center and from primary hyperoxaluria patients at the Mayo Clinic. The BMC were isolated and GR and DGDH activities measured in cell lysates. Results. An assay of 20 normal individuals indicated that BMC contained a DGDH and GR activity of 0.97±0.20 (range 0.62-1.45), and 10.6±3.3 (range 8.3-16.6) nmol/min/mg protein, respectively. The intra-assay coefficient of variation for DGDH and GR activity was 8.2 and 11.5{\%}, respectively. The BMC lysates from al adult subjects and patients with PH1 showed similar GR and DGDH activities. This was confirmed by the presence of immunoreactive GRHPR protein by western blot analysis. In contrast, PH2 BMC lysates did not exhibit DGDH or GR activity, and showed no immunoreactive GRHPR by western blot analysis. Conclusion. These results suggest that the assay of DGDH or GR activity in BMC could be used as a minimally invasive diagnostic test for PH2.",
keywords = "Blood mononuclear cells, D-glycerate dehydrogenase, Glyoxylate reductase, Primary hyperoxaluria type 2",
author = "John Knight and Holmes, {Ross P.} and Milliner, {Dawn S.} and Monico, {Carla G.} and Cramer, {Scott D.}",
year = "2006",
month = "8",
doi = "10.1093/ndt/gfl142",
language = "English (US)",
volume = "21",
pages = "2292--2295",
journal = "Nephrology Dialysis Transplantation",
issn = "0931-0509",
publisher = "Oxford University Press",
number = "8",

}

TY - JOUR

T1 - Glyoxylate reductase activity in blood mononuclear cells and the diagnosis of primary hyperoxaluria type 2

AU - Knight, John

AU - Holmes, Ross P.

AU - Milliner, Dawn S.

AU - Monico, Carla G.

AU - Cramer, Scott D.

PY - 2006/8

Y1 - 2006/8

N2 - Background. Primary hyperoxaluria type 2 (PH2) is a rare monogenic disorder characterized by an elevated urinary excretion of oxalate. Increased oxalate excretion in PH2 patients can cause nephrolithiasis and nephrocalcinosis, and can, in some cases, result in renal failure and systemic oxalate deposition. The disease is due to a deficiency of glyoxylate reductase/hydroxypyruvate reductase (GRHPR) activity. A definitive diagnosis of PH2 is currently made by the analysis of GR activity in a liver biopsy. GRHPR is expressed in virtually every tissue in the body, suggesting that utilization of more readily available cells could be used to determine GRHPR deficiency. In this study, we have evaluated the potential of determining GR and d -glycerate dehydrogenase (DGDH) activity in blood mononuclear cells (BMC) as a diagnostic indicator of PH2. Methods. Blood samples were obtained from 10 male and 10 female normal subjects, median age 31, range 21-63, at the Wake Forest University Medical Center and from primary hyperoxaluria patients at the Mayo Clinic. The BMC were isolated and GR and DGDH activities measured in cell lysates. Results. An assay of 20 normal individuals indicated that BMC contained a DGDH and GR activity of 0.97±0.20 (range 0.62-1.45), and 10.6±3.3 (range 8.3-16.6) nmol/min/mg protein, respectively. The intra-assay coefficient of variation for DGDH and GR activity was 8.2 and 11.5%, respectively. The BMC lysates from al adult subjects and patients with PH1 showed similar GR and DGDH activities. This was confirmed by the presence of immunoreactive GRHPR protein by western blot analysis. In contrast, PH2 BMC lysates did not exhibit DGDH or GR activity, and showed no immunoreactive GRHPR by western blot analysis. Conclusion. These results suggest that the assay of DGDH or GR activity in BMC could be used as a minimally invasive diagnostic test for PH2.

AB - Background. Primary hyperoxaluria type 2 (PH2) is a rare monogenic disorder characterized by an elevated urinary excretion of oxalate. Increased oxalate excretion in PH2 patients can cause nephrolithiasis and nephrocalcinosis, and can, in some cases, result in renal failure and systemic oxalate deposition. The disease is due to a deficiency of glyoxylate reductase/hydroxypyruvate reductase (GRHPR) activity. A definitive diagnosis of PH2 is currently made by the analysis of GR activity in a liver biopsy. GRHPR is expressed in virtually every tissue in the body, suggesting that utilization of more readily available cells could be used to determine GRHPR deficiency. In this study, we have evaluated the potential of determining GR and d -glycerate dehydrogenase (DGDH) activity in blood mononuclear cells (BMC) as a diagnostic indicator of PH2. Methods. Blood samples were obtained from 10 male and 10 female normal subjects, median age 31, range 21-63, at the Wake Forest University Medical Center and from primary hyperoxaluria patients at the Mayo Clinic. The BMC were isolated and GR and DGDH activities measured in cell lysates. Results. An assay of 20 normal individuals indicated that BMC contained a DGDH and GR activity of 0.97±0.20 (range 0.62-1.45), and 10.6±3.3 (range 8.3-16.6) nmol/min/mg protein, respectively. The intra-assay coefficient of variation for DGDH and GR activity was 8.2 and 11.5%, respectively. The BMC lysates from al adult subjects and patients with PH1 showed similar GR and DGDH activities. This was confirmed by the presence of immunoreactive GRHPR protein by western blot analysis. In contrast, PH2 BMC lysates did not exhibit DGDH or GR activity, and showed no immunoreactive GRHPR by western blot analysis. Conclusion. These results suggest that the assay of DGDH or GR activity in BMC could be used as a minimally invasive diagnostic test for PH2.

KW - Blood mononuclear cells

KW - D-glycerate dehydrogenase

KW - Glyoxylate reductase

KW - Primary hyperoxaluria type 2

UR - http://www.scopus.com/inward/record.url?scp=33748059855&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=33748059855&partnerID=8YFLogxK

U2 - 10.1093/ndt/gfl142

DO - 10.1093/ndt/gfl142

M3 - Article

VL - 21

SP - 2292

EP - 2295

JO - Nephrology Dialysis Transplantation

JF - Nephrology Dialysis Transplantation

SN - 0931-0509

IS - 8

ER -