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Animal Models and "Omics" Technologies for Identification of Novel Biomarkers and Drug Targets to Prevent Heart Failure.

Hou Y, Adrian-Segarra JM, Richter M, Kubin N, Shin J, Werner I, Walther T, Schönburg M, Pöling J, Warnecke H, Braun T, Kostin S, Kubin T - Biomed Res Int (2015)

Bottom Line: Despite its complexity, stressed cardiomyocytes often follow conserved patterns of structural remodelling in order to adapt, survive, and regenerate.To achieve this, various animal models in combination with an "omics" toolbox can be used.These approaches will ultimately lead to the identification of an arsenal of biomarkers and therapeutic targets which have the potential to shape the medicine of the future.

View Article: PubMed Central - PubMed

Affiliation: Department of Cardiac Development and Remodelling, Max Planck Institute for Heart and Lung Research, Ludwigstrasse 43, 61231 Bad Nauheim, Germany.

ABSTRACT
It is now accepted that heart failure (HF) is a complex multifunctional disease rather than simply a hemodynamic dysfunction. Despite its complexity, stressed cardiomyocytes often follow conserved patterns of structural remodelling in order to adapt, survive, and regenerate. When cardiac adaptations cannot cope with mechanical, ischemic, and metabolic loads efficiently or become chronically activated, as, for example, after infection, then the ongoing structural remodelling and dedifferentiation often lead to compromised pump function and patient death. It is, therefore, of major importance to understand key events in the progression from a compensatory left ventricular (LV) systolic dysfunction to a decompensatory LV systolic dysfunction and HF. To achieve this, various animal models in combination with an "omics" toolbox can be used. These approaches will ultimately lead to the identification of an arsenal of biomarkers and therapeutic targets which have the potential to shape the medicine of the future.

No MeSH data available.


Related in: MedlinePlus

The design of an “omics” technology research platform consisting of complementary core facilities for the future of a personalized medicine (adapted and modified from the “Venia legendi” work of J. Pöling, 2013). (a) Multiplex systems, Western blot, and confocal microscopy are antibody based methods. The Western blot shows a stained membrane resolving 1 μL of two serum samples (2&3) and 1 μL of pericardial fluid (PCF) from a patient with HF and high myocardial level of oncostatin M (size marker was run parallel in 1). A strong signal of TIMP-1 was detected in PCF after antibody hybridization of the membrane and subsequent chemiluminescence detection (WB). When targets are not known the separation of proteins lysates by 2-dimensional gel electrophoresis (2DE) reveals thousands of not yet defined protein spots after silver staining (proteome analysis). Here, a cytoplasmic cardiac 2DE of a 3-day-old rat is shown. Then, gels are scanned and compared and a computer based software program identifies regulated spots. (4) The protein spot is excised and identified by mass spectrometry combined with database searches. (1) In addition RNA might be extracted from the same sample and expression levels of ten thousands of genes can be simultaneously analyzed on DNA microarrays (transcriptome analysis). (5) Confocal microscopy verifies observations and provides further information about the protein. A confocal image shows FGF23-positive cardiomyocytes in a patient with aortic stenosis and high myocardial level of oncostatin M. (b) ANP, BNP, interleukin-6, and FGF-23 were identified as oncostatin M-regulated biomarkers on this platform. Further potential biomarkers such as radixin and moesin are indicated.
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fig2: The design of an “omics” technology research platform consisting of complementary core facilities for the future of a personalized medicine (adapted and modified from the “Venia legendi” work of J. Pöling, 2013). (a) Multiplex systems, Western blot, and confocal microscopy are antibody based methods. The Western blot shows a stained membrane resolving 1 μL of two serum samples (2&3) and 1 μL of pericardial fluid (PCF) from a patient with HF and high myocardial level of oncostatin M (size marker was run parallel in 1). A strong signal of TIMP-1 was detected in PCF after antibody hybridization of the membrane and subsequent chemiluminescence detection (WB). When targets are not known the separation of proteins lysates by 2-dimensional gel electrophoresis (2DE) reveals thousands of not yet defined protein spots after silver staining (proteome analysis). Here, a cytoplasmic cardiac 2DE of a 3-day-old rat is shown. Then, gels are scanned and compared and a computer based software program identifies regulated spots. (4) The protein spot is excised and identified by mass spectrometry combined with database searches. (1) In addition RNA might be extracted from the same sample and expression levels of ten thousands of genes can be simultaneously analyzed on DNA microarrays (transcriptome analysis). (5) Confocal microscopy verifies observations and provides further information about the protein. A confocal image shows FGF23-positive cardiomyocytes in a patient with aortic stenosis and high myocardial level of oncostatin M. (b) ANP, BNP, interleukin-6, and FGF-23 were identified as oncostatin M-regulated biomarkers on this platform. Further potential biomarkers such as radixin and moesin are indicated.

Mentions: An ideal “omics” platform consists of complementary core facilities in order to obtain as much information as possible from fluids, cell cultures, and tissue samples (Figure 2(a)). Fluids are flush-frozen in liquid nitrogen after centrifugation and removal of insoluble material. Usually fluids do not need further processing and can be directly analysed by commercially available ELISA kits (single protein detection by specific antibodies in one sample) or kits using multiplex systems. Multiple cytokine and chemokine assays are multiplex bead-based assays able to simultaneously quantify up to 32 (or even more) targets in one sample. The main advantage of multiplex systems lies in the speed (data are obtained on the same day with little working effort) and sensitivity of determinations (down to picogram levels of proteins). Furthermore, a much lower amount of sample is needed compared to an ELISA due to simultaneous detection of multiple proteins. “Multiplexing” is especially powerful when using highly diluted samples, such as fluids, but might have limitations in the study of tissue biopsies that have to be homogenized in buffers containing detergents. Limitations are further seen in the number of commercially available kits containing panels of different antibodies recognizing predefined protein targets, and the specificity and accuracy of these multiplex systems might be sometimes a matter of concern.


Animal Models and "Omics" Technologies for Identification of Novel Biomarkers and Drug Targets to Prevent Heart Failure.

Hou Y, Adrian-Segarra JM, Richter M, Kubin N, Shin J, Werner I, Walther T, Schönburg M, Pöling J, Warnecke H, Braun T, Kostin S, Kubin T - Biomed Res Int (2015)

The design of an “omics” technology research platform consisting of complementary core facilities for the future of a personalized medicine (adapted and modified from the “Venia legendi” work of J. Pöling, 2013). (a) Multiplex systems, Western blot, and confocal microscopy are antibody based methods. The Western blot shows a stained membrane resolving 1 μL of two serum samples (2&3) and 1 μL of pericardial fluid (PCF) from a patient with HF and high myocardial level of oncostatin M (size marker was run parallel in 1). A strong signal of TIMP-1 was detected in PCF after antibody hybridization of the membrane and subsequent chemiluminescence detection (WB). When targets are not known the separation of proteins lysates by 2-dimensional gel electrophoresis (2DE) reveals thousands of not yet defined protein spots after silver staining (proteome analysis). Here, a cytoplasmic cardiac 2DE of a 3-day-old rat is shown. Then, gels are scanned and compared and a computer based software program identifies regulated spots. (4) The protein spot is excised and identified by mass spectrometry combined with database searches. (1) In addition RNA might be extracted from the same sample and expression levels of ten thousands of genes can be simultaneously analyzed on DNA microarrays (transcriptome analysis). (5) Confocal microscopy verifies observations and provides further information about the protein. A confocal image shows FGF23-positive cardiomyocytes in a patient with aortic stenosis and high myocardial level of oncostatin M. (b) ANP, BNP, interleukin-6, and FGF-23 were identified as oncostatin M-regulated biomarkers on this platform. Further potential biomarkers such as radixin and moesin are indicated.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4508378&req=5

fig2: The design of an “omics” technology research platform consisting of complementary core facilities for the future of a personalized medicine (adapted and modified from the “Venia legendi” work of J. Pöling, 2013). (a) Multiplex systems, Western blot, and confocal microscopy are antibody based methods. The Western blot shows a stained membrane resolving 1 μL of two serum samples (2&3) and 1 μL of pericardial fluid (PCF) from a patient with HF and high myocardial level of oncostatin M (size marker was run parallel in 1). A strong signal of TIMP-1 was detected in PCF after antibody hybridization of the membrane and subsequent chemiluminescence detection (WB). When targets are not known the separation of proteins lysates by 2-dimensional gel electrophoresis (2DE) reveals thousands of not yet defined protein spots after silver staining (proteome analysis). Here, a cytoplasmic cardiac 2DE of a 3-day-old rat is shown. Then, gels are scanned and compared and a computer based software program identifies regulated spots. (4) The protein spot is excised and identified by mass spectrometry combined with database searches. (1) In addition RNA might be extracted from the same sample and expression levels of ten thousands of genes can be simultaneously analyzed on DNA microarrays (transcriptome analysis). (5) Confocal microscopy verifies observations and provides further information about the protein. A confocal image shows FGF23-positive cardiomyocytes in a patient with aortic stenosis and high myocardial level of oncostatin M. (b) ANP, BNP, interleukin-6, and FGF-23 were identified as oncostatin M-regulated biomarkers on this platform. Further potential biomarkers such as radixin and moesin are indicated.
Mentions: An ideal “omics” platform consists of complementary core facilities in order to obtain as much information as possible from fluids, cell cultures, and tissue samples (Figure 2(a)). Fluids are flush-frozen in liquid nitrogen after centrifugation and removal of insoluble material. Usually fluids do not need further processing and can be directly analysed by commercially available ELISA kits (single protein detection by specific antibodies in one sample) or kits using multiplex systems. Multiple cytokine and chemokine assays are multiplex bead-based assays able to simultaneously quantify up to 32 (or even more) targets in one sample. The main advantage of multiplex systems lies in the speed (data are obtained on the same day with little working effort) and sensitivity of determinations (down to picogram levels of proteins). Furthermore, a much lower amount of sample is needed compared to an ELISA due to simultaneous detection of multiple proteins. “Multiplexing” is especially powerful when using highly diluted samples, such as fluids, but might have limitations in the study of tissue biopsies that have to be homogenized in buffers containing detergents. Limitations are further seen in the number of commercially available kits containing panels of different antibodies recognizing predefined protein targets, and the specificity and accuracy of these multiplex systems might be sometimes a matter of concern.

Bottom Line: Despite its complexity, stressed cardiomyocytes often follow conserved patterns of structural remodelling in order to adapt, survive, and regenerate.To achieve this, various animal models in combination with an "omics" toolbox can be used.These approaches will ultimately lead to the identification of an arsenal of biomarkers and therapeutic targets which have the potential to shape the medicine of the future.

View Article: PubMed Central - PubMed

Affiliation: Department of Cardiac Development and Remodelling, Max Planck Institute for Heart and Lung Research, Ludwigstrasse 43, 61231 Bad Nauheim, Germany.

ABSTRACT
It is now accepted that heart failure (HF) is a complex multifunctional disease rather than simply a hemodynamic dysfunction. Despite its complexity, stressed cardiomyocytes often follow conserved patterns of structural remodelling in order to adapt, survive, and regenerate. When cardiac adaptations cannot cope with mechanical, ischemic, and metabolic loads efficiently or become chronically activated, as, for example, after infection, then the ongoing structural remodelling and dedifferentiation often lead to compromised pump function and patient death. It is, therefore, of major importance to understand key events in the progression from a compensatory left ventricular (LV) systolic dysfunction to a decompensatory LV systolic dysfunction and HF. To achieve this, various animal models in combination with an "omics" toolbox can be used. These approaches will ultimately lead to the identification of an arsenal of biomarkers and therapeutic targets which have the potential to shape the medicine of the future.

No MeSH data available.


Related in: MedlinePlus