Academic Senate - Meeting Minutes, 4/18/2017

<p>All values are presented with SD. Differences between <i>LDLR−/−</i> and the other two genotypes are significant where indicated, ANOVA: *p<0.05, **p<0.01.</p

Weekly change in % of lesion score.

<p>The data in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0132092#pone.0132092.g001" target="_blank">Fig 1</a> is shown as the mean rate of change for each treatment. Data are figured as the % change per week in Lesion score. Means with the same subscript do not differ significantly. Both NAC and GSH treated animals showed a significant decrease in lesion scores over time.</p

Antioxidant Therapies for Ulcerative Dermatitis: A Potential Model for Skin Picking Disorder

<div><p>Skin Picking Disorder affects 4% of the general population, with serious quality of life impacts, and potentially life threatening complications. Standard psychoactive medications do not help most patients. Similarly, Mouse Ulcerative Dermatitis (skin lesions caused by excessive abnormal grooming behavior) is very common in widely used inbred strains of mice, and represents a serious animal welfare issue and cause of mortality. Treatment options for Ulcerative Dermatitis are largely palliative and ineffective. We have proposed mouse Ulcerative Dermatitis as a model for human Skin Picking Disorder based on similar epidemiology, behavior, and its comorbidity and mechanistic overlap with hair pulling (trichotillomania). We predicted that mouse Ulcerative Dermatitis would be treated by N-Acetylcysteine, as this compound is highly effective in treating both Skin Picking Disorder and Trichotillomania. Furthermore, we hypothesized that N-Acetylcysteine’s mode of action is as a precursor to the production of the endogenous antioxidant glutathione in the brain, and therefore intranasal glutathione would also treat Ulcerative Dermatitis. Accordingly, we show in a heterogenous prospective trial, the significant reduction in Ulcerative Dermatitis lesion severity in mice receiving either N-acetylcysteine (oral administration) or glutathione (intranasal). The majority of mice treated with N-acetylcysteine improved slowly throughout the course of the study. Roughly half of the mice treated with glutathione showed complete resolution of lesion within 2-4 weeks, while the remainder did not respond. These findings are the first to show that the use of N-acetylcysteine and Glutathione can be curative for mouse Ulcerative Dermatitis. These findings lend additional support for mouse Ulcerative Dermatitis as a model of Skin Picking Disorder and also support oxidative stress and glutathione synthesis as the mechanism of action for these compounds. As N-Acetylcysteine is poorly tolerated by many patients, intranasal glutathione warrants further study as potential therapy in Skin Picking, trichotillomania and other body-focused repetitive behavior disorders.</p></div

Mean time to Cure.

<p>For animals that were completely cured (i.e. had final UD lesions scores of zero), GSH achieved this outcome significantly faster than NAC.</p

Mean UD lesion score as a function of time.

<p>Probands did not differ in mean score at week 0. However, cagemates of probands who later developed UD are included, reducing the mean score for ‘None’ animals at week 0. Data are shown as the arithmetic mean +/- SE of the lesion scores corrected for subject, averaged at each week (horizontal error bars show the SE of the day of observation). Post-hoc tests of the slopes are shown in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0132092#pone.0132092.g002" target="_blank">Fig 2</a>.</p

Ulcerative Dermatitis Lesion Severity Scale.

<p>Lesions were graded for severity on a five point scale as shown, and drawn on a standardized ‘mouse map’. The UD score for each mouse was calculated as the area of the lesion weighted by the severity.</p

Percentage of Mice Cured Over Time.

<p>The raw event data behind <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0132092#pone.0132092.g003" target="_blank">Fig 3</a> are shown. Euthanasia and cure events cannot be show on the same graph, however the final percentage of animals cured also reflects the cumulative proportion of animals euthanized by the end of the study (i.e. every non-cured animal was euthanized at some point during the study according to our human endpoints).</p

Summary of <i>LDLR</i> gene targeting and SCNT activity.

<p>*Gene targeting efficiency reported as percentage of G418^R clones that were properly targeted, as determined by PCR.</p>†<p>Pregnancy rate refers to full-term gestation.</p

Fasting Insulin Levels and Bergman Si Values.

<p>*Wilcoxon signed rank statistic for change from baseline within group.</p><p>^†Wilcoxon rank sum statistic for differences between groups.</p><p>Fasting Insulin Levels and Bergman Si Values.</p

Targeted Disruption of <i>LDLR</i> Causes Hypercholesterolemia and Atherosclerosis in Yucatan Miniature Pigs

<div><p>Recent progress in engineering the genomes of large animals has spurred increased interest in developing better animal models for diseases where current options are inadequate. Here, we report the creation of Yucatan miniature pigs with targeted disruptions of the low-density lipoprotein receptor (<i>LDLR</i>) gene in an effort to provide an improved large animal model of familial hypercholesterolemia and atherosclerosis. Yucatan miniature pigs are well established as translational research models because of similarities to humans in physiology, anatomy, genetics, and size. Using recombinant adeno-associated virus-mediated gene targeting and somatic cell nuclear transfer, male and female <i>LDLR+/−</i> pigs were generated. Subsequent breeding of heterozygotes produced <i>LDLR−/−</i> pigs. When fed a standard swine diet (low fat, no cholesterol), <i>LDLR+/−</i> pigs exhibited a moderate, but consistent increase in total and LDL cholesterol, while <i>LDLR−/−</i> pigs had considerably elevated levels. This severe hypercholesterolemia in homozygote animals resulted in atherosclerotic lesions in the coronary arteries and abdominal aorta that resemble human atherosclerosis. These phenotypes were more severe and developed over a shorter time when fed a diet containing natural sources of fat and cholesterol. <i>LDLR</i>-targeted Yucatan miniature pigs offer several advantages over existing large animal models including size, consistency, availability, and versatility. This new model of cardiovascular disease could be an important resource for developing and testing novel detection and treatment strategies for coronary and aortic atherosclerosis and its complications.</p></div