Difference between revisions of "Krylova 2017 MiPschool Obergurgl"

Revision as of 13:18, 10 July 2017

High-resolution respirometry in diagnostic of mitochondrial complex I deficiency.

Link: MITOEAGLE

Krylova TD, Tsygankova PG, Itkis YS, Sheremet NL, Nevinitsyna TA, Mikhaylova SV, Zakharova EY (2017)

Event: MiPschool Obergurgl 2017

Complex I (CI) deficiency is one of the most common defects in OXPHOS system and representing more than 30% cases of mitochondrial diseases. The group is characterized by clinical and genetic heterogeneity and comprise several nosological forms. The most prevalent phenotypes for complex I deficiency are LHON and Leigh syndrome.

In this study we analyzed skin fibroblasts from 11 patients with mutations in mtDNA, which cause LHON or Leigh-like phenotypes: m.11778 G>A (N=3), m.3460 A>G (N=2), m.3635 G>A (N=1), m.3308 T>G (N=2), m.3472 T>C (N=1) and 2 patients with unpublished substitutions m.3945 C>A and m.14441T>C. High-resolution respirometry (HRR) on the Oxygraph-2k (OROBOROS INSTRUMENTS, Austria) was performed for complex analysis of mitochondrial respiratory function in intact and permeabilized fibroblasts of patients and healthy controls.

Flux control rations in intact cells R/E, (R-L)/E (p<0,05) were raised compared to the control. Rates R, E, L normalized on the citrate synthase activity (CS) were statistically varied between patients and controls. In permeabilized fibroblasts we observed differences in СII/E, Rot/E, R/CII, CI/CII (p<0,05) between groups. These data highlight the dysfunction of OXPHOS system and particularly CI. Increased CS level and decreased CI/CII ratio indicate compensatory metabolic answer on respiratory chain dysfunction.

Our results show the facilities of HRR in revealing the biochemical abnormalities of complex I in patient’s fibroblasts with LHON and Leigh-like syndrome. We also suggest HRR to be useful method for inspection other mutations causing complex I deficiency.

• Bioblast editor: Kandolf G

Labels: MiParea: Respiration, mtDNA;mt-genetics, mt-Medicine

Tissue;cell: Endothelial;epithelial;mesothelial cell, Fibroblast Preparation: Intact cells, Permeabilized cells

Coupling state: LEAK, ROUTINE, ETS"ETS" is not in the list (LEAK, ROUTINE, OXPHOS, ET) of allowed values for the "Coupling states" property. Pathway: N, S HRR: Oxygraph-2k

Affiliations

(1)Krylova TD, (1)Tsygankova PG, (1)Itkis YS, (2)Sheremet NL, (2)Nevinitsyna TA, (3)Mikhaylova SV, (1)Zakharova EY1

Res Centre Med Gen
Res Inst Eye Dis
Rus Child Clin Hosp

Moscow, Russia. – [email protected]

@@ Line 1: / Line 1: @@
 {{Abstract
-|title=[[File:MITOEAGLE-representation.jpg|left|60px|link=http://www.mitoglobal.org/index.php/MITOEAGLE|COST Action MITOEAGLE]]
+|title=[[File:MITOEAGLE-representation.jpg|left|60px|link=http://www.mitoglobal.org/index.php/MITOEAGLE|COST Action MITOEAGLE]] High-resolution respirometry in diagnostic of mitochondrial complex I deficiency.
 |info=[[MITOEAGLE]]
+|authors=Krylova TD, Tsygankova PG, Itkis YS, Sheremet NL, Nevinitsyna TA, Mikhaylova SV, Zakharova EY
 |year=2017
 |event=MiPschool Obergurgl 2017
 |abstract=[[Image:MITOEAGLE-logo.jpg|left|100px|link=http://www.mitoglobal.org/index.php/MITOEAGLE|COST Action MITOEAGLE]]
+Complex I (CI) deficiency is one of the most common defects in OXPHOS system and representing more than 30% cases of mitochondrial diseases. The group is characterized by clinical and genetic heterogeneity and comprise several nosological forms. The most prevalent phenotypes for complex I deficiency are LHON and Leigh syndrome.
+In this study we analyzed skin fibroblasts from 11 patients with mutations in mtDNA, which cause LHON or Leigh-like phenotypes: m.11778 G>A (N=3), m.3460 A>G (N=2), m.3635 G>A (N=1), m.3308 T>G (N=2), m.3472 T>C (N=1) and 2 patients with unpublished substitutions m.3945 C>A and m.14441T>C. High-resolution respirometry (HRR) on the Oxygraph-2k (OROBOROS INSTRUMENTS, Austria) was performed for complex analysis of mitochondrial respiratory function in intact and permeabilized fibroblasts of patients and healthy controls.
+Flux control rations in intact cells R/E, (R-L)/E (p<0,05) were raised compared to the control. Rates R, E, L normalized on the citrate synthase activity (CS) were statistically varied between patients and controls. In permeabilized fibroblasts we observed differences in СII/E, Rot/E, R/CII, CI/CII (p<0,05) between groups. These data highlight the dysfunction of OXPHOS system and particularly CI. Increased CS level and decreased CI/CII ratio indicate compensatory metabolic answer on respiratory chain dysfunction.
+Our results show the facilities of HRR in revealing the biochemical abnormalities of complex I in patient’s fibroblasts with LHON and Leigh-like syndrome. We also suggest HRR to be useful method for inspection other mutations causing complex I deficiency.
+|editor=[[Kandolf G]],
+}}
+{{Labeling
+|area=Respiration, mtDNA;mt-genetics, mt-Medicine
+|tissues=Endothelial;epithelial;mesothelial cell, Fibroblast
+|preparations=Intact cells, Permeabilized cells
+|couplingstates=LEAK, ROUTINE, ETS
+|pathways=N, S
+|instruments=Oxygraph-2k
 }}
-{{Labeling}}
 == Affiliations ==
-:::: (1)
+:::: (1)Krylova TD, (1)Tsygankova PG, (1)Itkis YS, (2)Sheremet NL, (2)Nevinitsyna TA, (3)Mikhaylova SV, (1)Zakharova EY1
+::::#Res Centre Med Gen
-::::#
+::::#Res Inst Eye Dis
+::::#Rus Child Clin Hosp
-== Figure 1 ==
+::::::Moscow, Russia. – [email protected]