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Kappler 2018 J Lab Med

From Bioblast
Kappler L, Hoene M, Kollipara L, Li J, Hu C, van Törne C, Hauck S, Irmler M, Beckers J, de Angelis MH, Peter A, Xu G, Häring H, Weigert C, Sickmann A (2018) Systems medicine approach combining Multi-omics and functional analyses in hepatic and skeletal muscle mitochondria of pre-diabetic mice. J Lab Med.

Link: German Congress of Laboratory Medicine

Kappler L, Hoene M, Kollipara L, Li J, Hu C, van Toerne C, Hauck S, Irmler M, Beckers J, de Angelis MH, Peter A, Xu G, Haering H, Weigert C, Sickmann A (2018)

Event: J Lab Med

Mitochondria are discussed as one of the key players in the pathogenesis of type 2 diabetes. However, experimental evidence is controversial, pointing towards a mitochondrial dysfunction associated with insulin resistance, but also towards compensatory mechanisms.

Comprehensive investigation of mitochondrial adaptions to high-energy diet (HED) by a multi-omics approach and functional studies to provide new insights into diabetes-associated pathomechanisms in a tissue-specific mitochondrial manner.

We applied a systems medicine approach combining transcriptomics and lipidomics, proteomics and functional analyses of isolated mitochondria and total tissue to elucidate the interplay between molecular fingerprints and their impact on mitochondrial function. Purified mitochondria (by ultracentrifugation) from skeletal muscle and liver were investigated by LC-coupled proteomic/lipidomic analyses (Q Exactive HF) in a pre-diabetic mouse model (6 weeks of control (10 fat/20 protein/70 carbohydrates [kJ%]) vs high-energy diet (45 fat/20 protein/35 carbohydrates [kJ%]), n = 8). Mitochondrial proteome was covered by non targeted DIA. Mitochondrial function was investigated by enzymatic assays and high-resolution respirometry, results were validated in human samples. Whole liver transcriptomics was performed.

More than 22.200 transcripts in whole liver, 950 hepatic mitochondrial proteins and above 540 proteins in isolated muscle mitochondria, as well as around 280 mitochondrial lipid species in both tissue mitochondria were quantified and linked to mitochondrial function. Mitochondria from muscle and liver showed distinctly different lipid and protein profiles, and also different substrate preferences in respirometry. After facing the HED, fatty acid metabolism pathways were enriched, both observed on protein but also transcript level and in fatty acid oxidation in muscle. Various tissue-specific mitochondrial adaptions were observed (assumed to be rescue mechanisms) to avoid substrate flooding of mitochondria, lowering the electron pressure on the electron transport chain and thereby providing protection from cell damage. However, alterations on mitochondrial lipid profile point towards the breakdown of these rescue mechanisms over time.

We linked the mitochondrial molecular composition with tissue-specific mitochondrial (dys)function generating a novel systems medicine tool for elucidating mitochondrial adaptions to e.g. diet-induced insulin resistance. Besides expected mitochondrial alterations due to the diet such as a changed fatty acid metabolism, rather unexpected rescue mechanisms were observed, providing further insights into the pathological remodeling of mitochondria in type 2 diabetes development.


Bioblast editor: Plangger M O2k-Network Lab: DE Tuebingen Weigert C


Labels: MiParea: Respiration, Exercise physiology;nutrition;life style  Pathology: Diabetes 

Organism: Mouse  Tissue;cell: Skeletal muscle, Liver  Preparation: Isolated mitochondria 



HRR: Oxygraph-2k 


Affiliations

Kappler L(1), Hoene M(1), Kollipara L(2), Li J(3), Hu C(3), van Törne C(4), Hauck S(4), Irmler M(5), Beckers J(5), de Angelis MH(5), Peter A(1), Xu G(3), Häring H(1), Weigert C(1), Sickmann A

  1. Univ Hospital Tuebingen, Clinical Chemistry Central Lab, Div Clinical Chemistry Pathobiochemistry, Dept Diagnostic Lab Med, Tübingen, Germany
  2. Leibniz-Inst Analytic Sciences, Protein Dynamics, Dortmund, Germany
  3. Dalian Inst Chemical Physics, Chinese Academy Sciences, Key Lab Separation Science Analytical Chemistry, Dalian, China
  4. Helmholtz Center München, Research Unit Protein Science, Munich, Germany
  5. Helmholtz Center München, German Research Center Environmental Health, Inst Experimental Genetics, Munich ,Germany
  6. Leibniz-Inst Analytic Sciences, Protein Dynamics, Dortmund, Germany