Modeling human protein aggregation cardiomyopathy using murine induced pluripotent stem cells

Autor: Paul Cheng, Pattraranee Limphong, Dennis R. Winge, Katie A. Mitzelfelt, Elisabeth S. Christians, Deepak Srivastava, Qiang Liu, Huali Zhang, Michael Riedel, Ivor J. Benjamin, Kathryn N. Ivey, Graydon Taylor
Rok vydání: 2013
Předmět:
Cellular differentiation
Medical Biotechnology
Protein aggregation
Cardiovascular
Transgenic
Mice
Natriuretic Peptide
Brain
Myocyte
2.1 Biological and endogenous factors
Myocytes
Cardiac
alpha-Crystallins
Induced pluripotent stem cell
Cardiomyocytes
Stem Cell Research - Induced Pluripotent Stem Cell - Human
Reverse Transcriptase Polymerase Chain Reaction
Brain
Cell Differentiation
General Medicine
beta-Crystallins
Cardiac hypertrophy
Heart Disease
Cardiac
Atrial Natriuretic Factor
Genetically modified mouse
Cardiomyopathy
Transgene
1.1 Normal biological development and functioning
Induced Pluripotent Stem Cells
Clinical Sciences
Mice
Transgenic
Biology
Cell Line
Troponin T
Natriuretic Peptide
Heat shock protein
Genetics
Animals
Humans
Actin
Embryonic Stem Cells/Induced Pluripotent Stem (iPS) Cells
Myocytes
alpha B-Crystallin
Stem Cell Research - Induced Pluripotent Stem Cell
Cell Biology
Cardiomyopathy
Hypertrophic
Stem Cell Research
Molecular biology
Actins
Gene Expression Regulation
Hypertrophic
Biochemistry and Cell Biology
Protein Multimerization
Developmental Biology
Zdroj: Stem cells translational medicine, vol 2, iss 3
Limphong, P; Zhang, H; Christians, E; Liu, Q; Riedel, M; Ivey, K; et al.(2013). Modeling human protein aggregation cardiomyopathy using murine induced pluripotent stem cells. Stem Cells Translational Medicine, 2(3), 161-166. doi: 10.5966/sctm.2012-0073. UCSF: Retrieved from: http://www.escholarship.org/uc/item/5kh5r965
DOI: 10.5966/sctm.2012-0073.
Popis: Several mutations in αB-crystallin (CryAB), a heat shock protein with chaperone-like activities, are causally linked to skeletal and cardiac myopathies in humans. To better understand the underlying pathogenic mechanisms, we had previously generated transgenic (TG) mice expressing R120GCryAB, which recapitulated distinguishing features of the myopathic disorder (e.g., protein aggregates, hypertrophic cardiomyopathy). To determine whether induced pluripotent stem cell (iPSC)-derived cardiomyocytes, a new experimental approach for human disease modeling, would be relevant to aggregation-prone disorders, we decided to exploit the existing transgenic mouse model to derive iPSCs from tail tip fibroblasts. Several iPSC lines were generated from TG and non-TG mice and validated for pluripotency. TG iPSC-derived cardiomyocytes contained perinuclear aggregates positive for CryAB staining, whereas CryAB protein accumulated in both detergent-soluble and insoluble fractions. iPSC-derived cardiomyocytes identified by cardiac troponin T staining were significantly larger when expressing R120GCryAB at a high level in comparison with TG low expressor or non-TG cells. Expression of fetal genes such as atrial natriuretic factor, B-type natriuretic peptide, and α-skeletal α-actin, assessed by quantitative reverse transcription-polymerase chain reaction, were increased in TG cardiomyocytes compared with non-TG, indicating the activation of the hypertrophic genetic program in vitro. Our study demonstrates for the first time that differentiation of R120G iPSCs into cardiomyocytes causes protein aggregation and cellular hypertrophy, recapitulating in vitro key pathognomonic hallmarks found in both animal models and patients. Our findings pave the way for further studies exploiting this cell model system for mechanistic and therapeutic investigations. © AlphaMed Press.
Databáze: OpenAIRE
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