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Generating Double Knockout Mice to Model Genetic Intervention for Diabetic Cardiomyopathy in Humans

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Mouse Genetics

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1194))

Abstract

Diabetes is a rapidly increasing disease that enhances the chances of heart failure twofold to fourfold (as compared to age and sex matched nondiabetics) and becomes a leading cause of morbidity and mortality. There are two broad classifications of diabetes: type1 diabetes (T1D) and type2 diabetes (T2D). Several mice models mimic both T1D and T2D in humans. However, the genetic intervention to ameliorate diabetic cardiomyopathy in these mice often requires creating double knockout (DKO). In order to assess the therapeutic potential of a gene, that specific gene is either overexpressed (transgenic expression) or abrogated (knockout) in the diabetic mice. If the genetic mice model for diabetes is used, it is necessary to create DKO with transgenic/knockout of the target gene to investigate the specific role of that gene in pathological cardiac remodeling in diabetics. One of the important genes involved in extracellular matrix (ECM) remodeling in diabetes is matrix metalloproteinase-9 (Mmp9). Mmp9 is a collagenase that remains latent in healthy hearts but induced in diabetic hearts. Activated Mmp9 degrades extracellular matrix (ECM) and increases matrix turnover causing cardiac fibrosis that leads to heart failure. Insulin2 mutant (Ins2+/−) Akita is a genetic model for T1D that becomes diabetic spontaneously at the age of 3–4 weeks and show robust hyperglycemia at the age of 10–12 weeks. It is a chronic model of T1D. In Ins2+/− Akita, Mmp9 is induced. To investigate the specific role of Mmp9 in diabetic hearts, it is necessary to create diabetic mice where Mmp9 gene is deleted. Here, we describe the method to generate Ins2+/−/Mmp9−/− (DKO) mice to determine whether the abrogation of Mmp9 ameliorates diabetic cardiomyopathy.

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Acknowledgement

This work is supported by National Institute of Health grants HL-113281 and HL116205 to Paras Kumar Mishra.

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Author notes

    Authors and Affiliations

    1. Department of Cellular and Integrative Physiology, University of Nebraska Medical Center, 985850 Nebraska Medical Center, Omaha, NE, 68198, USA

      Vishalakshi Chavali, Shyam Sundar Nandi & Paras Kumar Mishra Ph.D.

    2. Stem Cell Regulation and Animal Aging Section, Basic Research Laboratory, National Cancer Institute, National Institutes of Health, Frederick, MD, 21702, USA

      Shree Ram Singh

    3. Department of Anesthesiology, University of Nebraska Medical Center, 984455 Nebraska Medical Center, Omaha, NE, 68198, USA

      Paras Kumar Mishra Ph.D.

    Authors
    1. Vishalakshi Chavali
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    2. Shyam Sundar Nandi
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    3. Shree Ram Singh
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    4. Paras Kumar Mishra Ph.D.
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    Corresponding author

    Correspondence to Paras Kumar Mishra Ph.D. .

    Editor information

    Editors and Affiliations

    1. Stem Cell Regulation and Animal Aging Section, Basic Research Laboratory,, National Cancer Institute, National Institutes of Health, Frederick, MD, USA

      Shree Ram Singh

    2. Genetically Engineered Mouse Modeling Core/Department of Molecular Virology, Immunology and Medical Genetics, Wexner Medical Center, Comprehensive Cancer Center, The Ohio State University, Columbus, OH, USA

      Vincenzo Coppola

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    © 2014 Springer Science+Business Media New York

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    Chavali, V., Nandi, S.S., Singh, S.R., Mishra, P.K. (2014). Generating Double Knockout Mice to Model Genetic Intervention for Diabetic Cardiomyopathy in Humans. In: Singh, S., Coppola, V. (eds) Mouse Genetics. Methods in Molecular Biology, vol 1194. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-1215-5_22

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    • DOI: https://doi.org/10.1007/978-1-4939-1215-5_22

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    • Publisher Name: Humana Press, New York, NY

    • Print ISBN: 978-1-4939-1214-8

    • Online ISBN: 978-1-4939-1215-5

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