The proposed studies will use a mouse model to investigate causes for two conditions, infertility and arthropathy. Infertility affects ~ 10% of couples, with about half of this related to male infertility. Abnormal formation of bone-like substances in tissues that are normally not calcified (arthropathy) interferes with tissue function limits motion, and is often painful. Forms of ectopic calcification range from heterotopic ossification (occurs with aging, following trauma, and in genetic and idiopathic forms), to age-related aortic stenosis, and aortic valve calcification (which is a prominent complication limiting surgical repair of the heart and cardiac valves). Understanding the process of ectopic calcification thus has significant potential therapeutic benefit. Mice with disruption of two genes involved in circadian rhythm generation, CLOCK and NPAS2, not only lose their circadian rhythms, but they are also infertile and developed a severe, age-related and progressive ectopic calcification. CLOCK and NPAS2 normally partner with BMAL1 in regulating circadian rhythms;BMAL1- deficient mice are also have reduced fertility and progressive arthropathy. These pathological phenotypes do not occur in animals lacking circadian rhythms due to other genetic lesions, indicating that it is disruption of the CLOCK/NPAS2:BMAL1 transcriptional complexes, likely unrelated to circadian clock function, that allows these pathologies to develop. Studies of animals with gene disruption throughout the body are complicated by the multiple potential sites of action of these genes, ranging from potential effects on reproductive endocrinology and behavior to defects in spermatogenesis (for infertility), and multiple potential cell types of the musculo-skeletal system (for arthropathy). We will thus use advanced genetics approaches to produce tissue-specific genetic lesions, to identify and isolate the cell types involved in fertiliy regulation and in arthropathy prevention, allowing assessment of the molecular events that lead to the pathological conditions. Our overall hypothesis is that local dysregulation of gene expression in animals lacking CLOCK and NPAS2 (or BMAL1) allows these pathological conditions (calcification in specialized but poorly defined cell types at the junction of cartilage and tendon to bone, and infertility). Our overall objective is to define the cell types involved using an unbiased, genetic approach in vivo, and then use these same approaches to label the cell types involved and to isolate them for molecular analysis at times when the genes are critically important for preventing these phenotypes. Identification of the cellular and molecular events occurring in this very reproducible, genetic model in mice should contribute to understanding and ultimately regulating fertility, and for understanding and ultimately preventing pathological ectopic calcification in humans.

Public Health Relevance

The proposed studies use a mouse model to investigate causes for two conditions, infertility and abnormal formation of bone-like substances in tissues that are normally not calcified (arthropathy). Infertility affects ~ 10% of couples, while inappropriate calcification interferes with tissue function, limits motion, and is painful, and is prevalent aging-related condition with significant health impact. Understanding the processes by which bony substances are deposited in soft tissues thus has significant potential therapeutic benefit. We will investigate a mouse model that has infertility and also age- dependent deposition of bony material in soft tissues, seeking to identify cell types affected and the genes that are abnormally expressed in this pathological process. Identification of the cellular and molecular events occurring in this very reproducible, genetic model in mice should contribute to understanding and ultimately improving treatment for these conditions in humans.

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Research Project (R01)
Project #
2R01NS056125-06
Application #
8506154
Study Section
Neuroendocrinology, Neuroimmunology, Rhythms and Sleep Study Section (NNRS)
Program Officer
He, Janet
Project Start
2006-06-01
Project End
2018-03-31
Budget Start
2013-04-01
Budget End
2014-03-31
Support Year
6
Fiscal Year
2013
Total Cost
$363,946
Indirect Cost
$145,196
Name
University of Massachusetts Medical School Worcester
Department
Biology
Type
Schools of Medicine
DUNS #
603847393
City
Worcester
State
MA
Country
United States
Zip Code
01655
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Dallmann, Robert; DeBruyne, Jason P; Weaver, David R (2011) Photic resetting and entrainment in CLOCK-deficient mice. J Biol Rhythms 26:390-401
Dallmann, Robert; Weaver, David R (2010) Altered body mass regulation in male mPeriod mutant mice on high-fat diet. Chronobiol Int 27:1317-28
Etchegaray, Jean-Pierre; Yu, Elizabeth A; Indic, Premananda et al. (2010) Casein kinase 1 delta (CK1delta) regulates period length of the mouse suprachiasmatic circadian clock in vitro. PLoS One 5:e10303
Etchegaray, Jean-Pierre; Machida, Kazuhiko K; Noton, Elizabeth et al. (2009) Casein kinase 1 delta regulates the pace of the mammalian circadian clock. Mol Cell Biol 29:3853-66
Keaney Jr, John F; Weaver, David R (2009) Vascular rhythms and adaptation: do your arteries know what time it is? Circulation 119:1463-6
Kim, Soo Mi; Power, Andrea; Brown, Timothy M et al. (2009) Deletion of the secretory vesicle proteins IA-2 and IA-2beta disrupts circadian rhythms of cardiovascular and physical activity. FASEB J 23:3226-32

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