Correction: The genetic consequences of dog breed formation-Accumulation of deleterious genetic variation and fixation of mutations associated with myxomatous mitral valve disease in cavalier King Charles spaniels.

[This corrects the article DOI: 10.1371/journal.pgen.1009726.].

The genetic consequences of dog breed formation-Accumulation of deleterious genetic variation and fixation of mutations associated with myxomatous mitral valve disease in cavalier King Charles spaniels.

Selective breeding for desirable traits in strictly controlled populations has generated an extraordinary diversity in canine morphology and behaviour, but has also led to loss of genetic variation and random entrapment of disease alleles. As a consequence, specific diseases are now prevalent in certain breeds, but whether the recent breeding practice led to an overall increase in genetic load remains unclear. Here we generate whole genome sequencing (WGS) data from 20 dogs per breed from eight breeds and document a ~10% rise in the number of derived alleles per genome at evolutionarily conserved sites in the heavily bottlenecked cavalier King Charles spaniel breed (cKCs) relative to in most breeds studied here. Our finding represents the first clear indication of a relative increase in levels of deleterious genetic variation in a specific breed, arguing that recent breeding practices probably were associated with an accumulation of genetic load in dogs. We then use the WGS data to identify candidate risk alleles for the most common cause for veterinary care in cKCs-the heart disease myxomatous mitral valve disease (MMVD). We verify a potential link to MMVD for candidate variants near the heart specific NEBL gene in a dachshund population and show that two of the NEBL candidate variants have regulatory potential in heart-derived cell lines and are associated with reduced NEBL isoform nebulette expression in papillary muscle (but not in mitral valve, nor in left ventricular wall). Alleles linked to reduced nebulette expression may hence predispose cKCs and other breeds to MMVD via loss of papillary muscle integrity.

Dental home care in dogs - a questionnaire study among Swedish dog owners, veterinarians and veterinary nurses.

BACKGROUND: Periodontal disease remains one of the dog's most common health issues, even though it is largely preventable by tooth brushing. Implementation of daily tooth brushing would not only improve animal welfare, but also reduce veterinary costs for the owner. There is a paucity of studies investigating attitudes, opinions and practices of dog owners, veterinarians, and veterinary nurses regarding preventative dental home care in dogs. The objective of this study was to investigate these parameters in Sweden, thereby providing a basis for improved prophylactic strategies. METHODS: Validated questionnaire surveys were distributed to all Swedish dog owners (n = 209,263), veterinarians (n = 3657) and veterinary nurses (n = 1650) with e-mail addresses in the national registry. The response rates were 32% for dog owners and veterinarians, and 38% for veterinary nurses. The survey questions concerned attitudes, opinions and practices regarding dental home care, including whether dog owners received information concerning dental home care or not, and if this information resulted in implementation. RESULTS: Attitudes, opinions and practices regarding dental home care are presented for Swedish dog owners, veterinarians, and veterinary nurses. A fundamental finding was that the absolute majority of Swedish dog owners do not perform adequate prophylactic dental home care. Considerable discrepancies were identified in the opinions of veterinary health practitioners and dog owners regarding attitudes towards dental home care and conveying of information. Several areas for improvement in the communication between dog owners and veterinary health practitioners concerning dental home care were identified. CONCLUSION: Our results illustrates the need for validated methods to increase dog owner compliance with dental home care recommendations. We also see a need of further education, regarding canine dental home care, among veterinarians, veterinary nurses, and dog owners. The results from this unique study constitute an important foundation for future development of prophylactic strategies, with the ultimate goal to improve dental health, and thereby animal welfare, in dogs.

The epidemiology of osteochondrosis in an insured Swedish dog population.

Osteochondrosis (OC) is a focal disturbance of endochondral ossification due to a failure of blood supply to the epiphyseal growth cartilage. In dogs, OC most commonly affects the shoulder joint, followed by the elbow, tarsal, and stifle joints. The condition is associated with clinical signs such as lameness and pain and the prognosis varies depending on the affected joint. Most epidemiologic studies of OC in dogs were performed over 20 years ago, and updated estimates of disease incidence are lacking. Therefore, the objectives of this study were to provide population-based estimates of the incidence rate, cause-specific mortality rate, and age at diagnosis of appendicular OC (AOC, including OC of the shoulder, elbow, stifle, and tarsal joints) and stifle and tarsal OC separately, using data from Agria Djurförsäkring in Sweden (2011-2016). Further, the study aimed to evaluate the risk of OC in subgroups divided by breed and sex and describe previous, concurrent, and subsequent diagnoses of the affected joint in dogs with stifle or tarsal joint OC. The study population included just over 600,000 dogs, of which 685 were affected by AOC. Stifle joint OC (n = 113) was more common than tarsal joint OC (n = 80). The incidence rate of AOC was 3.77 (95% confidence interval (CI): 3.49-4.07) cases per 10,000 dog-years at risk, while the incidence rate of stifle and joint tarsal OC was 0.64 (95% CI: 0.53-0.77) and 0.43 (95% CI: 0.34-0.54) cases per 10,000 dog-years at risk, respectively. All breeds at increased risk of AOC were large or giant, and male dogs had an increased risk of AOC compared to female dogs (RR 1.76, 95% CI: 1.50-2.07, p < 0.001). The median age at first diagnosis during the study period was 0.74 (0.32-11.5) years for AOC, 2.62 (0.45-8.82) years for stifle joint OC, and 0.73 (0.35-7.35) years for tarsal joint OC. Of the dogs with stifle or tarsal joint OC, 30.2% and 15.0% had a previous diagnosis of stifle/tarsal joint pain or other unspecific clinical signs, respectively, and 13.8% of the dogs with stifle joint OC suffered subsequent cruciate ligament rupture. Osteochondrosis was the most common reason for euthanasia in the affected dogs. In total, 77 dogs were euthanised due to AOC during the study period.

The epidemiology of patellar luxation in an insured Swedish dog population.

Patellar luxation (PL) is a common orthopaedic condition in dogs. This study aimed to evaluate the incidence and cause-specific mortality rate, age at diagnosis, and risk factors for medial PL (MPL), lateral PL (LPL), and bidirectional PL (BPL). Other diagnoses in dogs with PL were also explored. The study population included just over 600,000 dogs insured by Agria Pet Insurance in Sweden (2011-2016). There were 2726 dogs with PL. Medial patellar luxation affected 90 % of the dogs with PL, followed by LPL (5.9 %), BPL (2.4 %), and unspecified PL (1.6 %). The median age at first diagnosis during the study period was 2.8 years for MPL, 2.7 years for LPL, and 1.5 years for BPL. In total, 168 (6.2 %) of the dogs with PL had cruciate ligament rupture. There were substantial breed-specific differences in the risk of PL: almost all breeds at increased risk of MPL were small-sized, while several of the breeds at increased risk of LPL were large-sized. The breeds at high risk of BPL varied in size. Females had an increased risk of MPL (RR 1.2, 95 % CI: 1.1-1.3, p < 0.001) and a decreased risk of LPL (RR 0.72, 95 % CI: 0.51-1.0, p = 0.042) compared to males. In total, 116 dogs were euthanised due to PL and the breeds with the highest risk of PL-related euthanasia were the Pyrenean mountain dog, Dogue de Bordeaux, and German pinscher. The median age for PL-related euthanasia was 2.2 years.

The epidemiology of cruciate ligament rupture in an insured Swedish dog population.

Cruciate ligament rupture (CLR) is a common orthopedic disorder in dogs. The study objectives were to evaluate incidence rate (IR), cause-specific mortality rate (CSMR) and risk factors for CLR in insured dogs. A single cohort study of dogs insured in Agria Pet Insurance in Sweden (2011-2016) was performed. Age at diagnosis, IR, CSMR and relative risk (RR) for CLR was calculated overall and per breed. The cohort included just over 600,000 dogs. The IR of CLR was 23.8 (95% confidence interval, 23.1-24.6) cases per 10,000 DYAR. The breeds with highest RR of CLR were Boerboel and Dogo Canario, while the breeds with lowest RR were Standard Dachshund and Miniature Pinscher. Dogue de Bordeaux had highest RR of euthanasia due to CLR. The median age at veterinary care claim for CLR was 7.1 (range 0.3-16.0) years and 6.6 (0.3-12) years at life insurance settlement. Large and giant breeds were generally diagnosed and euthanized due to CLR at a younger age compared to smaller breeds. The majority of the breeds with increased RR of CLR diagnosis and CLR-related euthanasia were large or giant. A pattern of increasing size and decreasing age at diagnosis/CLR-related euthanasia was observed.

The epidemiology of stifle joint disease in an insured Swedish dog population.

BACKGROUND: Stifle joint diseases (SJD) are common in dogs and include a variety of diagnoses. The objective of the study was to provide an overview of the epidemiology of SJD in insured dogs. METHODS: An historical single cohort study of dogs insured in Agria Pet Insurance (2011-2016) in Sweden was performed. Incidence and relative risk (RR) of SJD was calculated for the whole dog population and for subgroups divided by breed, breed group and sex. RESULTS: The study population included almost 600,000 insured dogs (>1.7 million dog-years). Ninety-three different stifle joint diagnoses were reported in 9624 dogs, and the most common were cruciate ligament rupture and patellar luxation. The incidence of SJD was 55.4 cases per 10,000 dog-years at risk. Bulldog and boerboel had the highest RR of SJD. The breeds that accounted for the highest proportion of all SJD claimed dogs were mixed breed and Labrador retriever. Female dogs had a slightly increased RR compared with male dogs (RR 1.06, p = 0.006). The incidence increased yearly during the observation period. CONCLUSION: The study demonstrates breed-specific differences in incidence of SJD in dogs, which may be of importance for breeders, dog owners and veterinarians.

A novel canine reference genome resolves genomic architecture and uncovers transcript complexity.

We present GSD_1.0, a high-quality domestic dog reference genome with chromosome length scaffolds and contiguity increased 55-fold over CanFam3.1. Annotation with generated and existing long and short read RNA-seq, miRNA-seq and ATAC-seq, revealed that 32.1% of lifted over CanFam3.1 gaps harboured previously hidden functional elements, including promoters, genes and miRNAs in GSD_1.0. A catalogue of canine "dark" regions was made to facilitate mapping rescue. Alignment in these regions is difficult, but we demonstrate that they harbour trait-associated variation. Key genomic regions were completed, including the Dog Leucocyte Antigen (DLA), T Cell Receptor (TCR) and 366 COSMIC cancer genes. 10x linked-read sequencing of 27 dogs (19 breeds) uncovered 22.1 million SNPs, indels and larger structural variants. Subsequent intersection with protein coding genes showed that 1.4% of these could directly influence gene products, and so provide a source of normal or aberrant phenotypic modifications.

Professional dental cleaning in dogs: clinical routines among Swedish veterinarians and veterinary nurses.

BACKGROUND: Dental disease is very common in dogs and veterinary professional dental cleaning and examination, together with daily dental home care, is the foundation for good dental health. To our knowledge, no previous study has investigated professional dental cleaning routines in small animal veterinary practice. A validated questionnaire survey was distributed to all veterinarians and veterinary nurses with registered e-mail addresses in the Swedish national registry (veterinarians; n = 3657, veterinary nurses; n = 1650). Response rates were 32% for veterinarians (V) and 38% for veterinary nurses (VN). RESULTS: In total, 73% (V)/96% (VN) of respondents reported that professional dental cleaning was performed at their work place under general anesthesia, and 27% (V)/18% (VN) that dental cleaning was performed under sedation. Of the respondents, 43% (V)/96% (VN) considered regular dental cleaning under general anesthesia fairly or very important, and 49% (V)/47% (VN) stated that it was sometimes important for good dental health in dogs. A majority of respondents, 84% (V)/97% (VN), reported that dental extractions were performed at their clinic, and 72% (V)/90% (VN) had access to dental radiography equipment. CONCLUSION: A majority of Swedish veterinarians and veterinary nurses perform professional dental cleaning under general anesthesia with access to dental radiography equipment, in accordance with national and international recommendations. However, a considerable proportion of professional dental cleanings were performed under sedation only, and extractions performed without access to dental radiography equipment were common, suggesting several areas of improvement in the routines in Swedish veterinary clinics and hospitals. Our results clearly indicate the need for improved educational efforts to increase the awareness among veterinary health professionals regarding guidelines and official recommendations in canine dental care.

Disease-related and overall survival in dogs with cranial cruciate ligament disease, a historical cohort study.

OBJECTIVE: To analyse the effect of treatment method and other risk factors on survival in dogs with cranial cruciate ligament disease (CCLD). METHODS: A historical cohort study of 333 dogs presenting with CCLD at two University Hospitals (2011-2016) was performed. Signalment, history, treatment and follow-up details were retrieved from medical records, dog owners and referring veterinarians. Treatment groups were defined; conservative or surgical with either lateral fabellotibial suture (LFS) or osteotomy procedures. Multivariable Cox proportional hazards models were applied to evaluate risk factors for disease-related and overall survival. RESULTS: Sixty-five dogs were conservatively managed, 125 treated with LFS and 143 with osteotomy techniques. At follow-up (autumn 2018), 164 dogs (49.3 %) were alive and 169 (50.7 %) were dead. Both final Cox proportional hazards models included variables for treatment, age, weight and hospital. In addition, the final disease-related model included a variable for orthopaedic comorbidity, while non-orthopaedic comorbidities and a time-varying effect for age on a linear scale were included in the overall survival model. Treatment method was found to have an effect on both disease-related and overall survival and surgical treatment was associated with a lower hazard than conservative treatment. CONCLUSION: Survival in dogs with CCLD is influenced by treatment strategy, comorbidities, age and weight.

Dog Owners' Perspectives on Canine Dental Health-A Questionnaire Study in Sweden.

Periodontal disease is one of the most common diseases affecting dogs, with a reported prevalence of at least 80% in dogs over 3 years of age. However, there is a lack of studies regarding dog owners' assessment of their dog's dental health, and whether they perceive clinical signs often associated with periodontal disease, i.e., dental calculus, halitosis or mobile or lost teeth. A validated questionnaire survey was distributed to all Swedish dog owners with email addresses in the national registry (n = 209,263). The response rate was 32%. The survey questions concerned opinions and practices regarding canine dental health, including assessment of dental health parameters and dog owners' ability to examine their dog's mouth. A construct (α = 0.76) was used to investigate dog owners' assessed symptoms of their dog's dental health in relation to background factors. Half of the respondents rated their dog's dental health as very good. However, one in four dog owners experienced difficulties when inspecting the dog's teeth. The most common reason for this difficulty was stated to be an uncooperative dog. Almost half of the dog owners reported halitosis to some degree in their dog, and almost four in ten owners reported dental calculus. One in eight dogs had been previously anesthetized for dental cleaning, and one in 12 dogs had experienced problems with gum disease, according to the owners. Owners' assessment varied significantly with the dog's age, weight, breed, breed group, sex, and concurrent disease. Owner-related factors that influenced the assessment of the dog's dental health were age, gender, education, county (urban/rural), and whether they were breeders or not. Dog owners with smaller dogs, older dogs and certain breeds predisposed to periodontal disease assessed their dog's dental health as worse than their counterparts, which is in agreement with previously reported higher prevalence of dental disease in these groups. This indicates that dog owners are able to perform relative assessment of their dog's dental health status. Our results also highlight the need for routine professional assessment of periodontal health, as well as education of dog owners and training of dogs to accept dental care procedures.

Development and validation of two questionnaires: Dental home care and dental health in Swedish dogs.

BACKGROUND: Dental disease is one of the most common health problems in dogs. However, no studies have investigated the attitudes, opinions and practices of dog owners, veterinarians and veterinary nurses regarding dental health and preventative dental home care in dogs. The objective of this study was therefore to develop and validate questionnaires for this purpose, in accordance with survey methodology guidelines. METHODS: Questionnaire items were determined based on the authors' academic knowledge and clinical experience, and modified throughout the validation process. Several measures were taken to reduce sampling, coverage, measurement and non-response errors. Content validity was assessed by Subject-Matter Experts (SME) and cognitive interviews were conducted in accordance with the "think-aloud protocol". Non-response analysis was performed using several methods. RESULTS: Constructs were identified using exploratory factor analysis and two predefined constructs from the dog owner questionnaire were confirmed "Dog owners' attitudes towards brushing their dog's teeth" (Cronbach's α = 0.86) and "Dog owners' assessment of their dog's dental health" (α = 0.76). Additionally, exploratory factor analysis identified three potential constructs. In the veterinary health practitioner questionnaire, two constructs were identified: "Veterinary health practitioners' attitudes towards dental chews and dental feed" (α = 0.78) and "Veterinary health practitioners' attitudes and opinions on dental problems and dental cleaning" (α = 0.73). Non-response analysis showed a higher proportion of women in the sample of dog owners and veterinarians compared to the target populations. Veterinarians in the sample were also younger. In contrast, gender and age distributions in veterinary nurses did not differ between sample and target. CONCLUSION: The validation presented in this work showed that the developed questionnaires could be used as accurate and reliable tools for measuring attitudes and practices regarding dental home care in dogs among Swedish dog owners, veterinarians and veterinary nurses.

Prognostic factors for outcome after transsphenoidal hypophysectomy in dogs with pituitary-dependent hyperadrenocorticism.

OBJECT: The aim of this study was to determine prognostic factors for outcome after transsphenoidal hypophysectomy in dogs with pituitary-dependent hyperadrenocorticism (PDH). METHODS: One veterinary neurosurgeon performed transsphenoidal hypophysectomies in 181 dogs with PDH over a 12-year period. Survival analysis was performed with the Kaplan-Meier method. Prognostic factors were analyzed with the univariate Cox proportional hazard analysis followed by stepwise multivariate analysis. The log-rank test was used to assess disease-free fractions in three groups categorized according to early postoperative urinary corticoid/creatinine (C/C) ratios. RESULTS: Multivariate analysis revealed that old age, large pituitary size, and high preoperative concentrations of plasma adrenocorticotropic hormone were associated with an increased risk of PDH-related death. In addition, large pituitary size, thick sphenoid bone, high C/C ratio, and high concentration of plasma alpha-melanocyte-stimulating hormone (alpha-MSH) before surgery were associated with an increased risk of disease recurrence in the dogs that went into remission after hypophysectomy. Disease-free fractions were significantly higher in dogs with postoperative urinary C/C ratios in the lower normal range (< 5 x 10(-6)) than in dogs with postoperative C/C ratios in the upper normal range (5-10 x 10(-6)). CONCLUSIONS: The results of this study indicate that pituitary size, sphenoid bone thickness, plasma alpha-MSH concentration, and preoperative level of urinary cortisol excretion are predictors of long-term remission after transsphenoidal hypophysectomy for PDH in dogs. Urinary C/C ratios measured 6 to 10 weeks after surgery can be used as a guide for predicting the risk of tumor recurrence.

Catestatin, vasostatin, cortisol, and pain assessments in dogs suffering from traumatic bone fractures.

BACKGROUND: Traumatic bone fractures cause moderate to severe pain, which needs to be minimized for optimal recovery and animal welfare, illustrating the need for reliable objective pain biomarkers for use in a clinical setting. The objectives of this study were to investigate catestatin (CST) and vasostatin (VS) concentrations as two new potential biomarkers, and cortisol concentrations, scores of the short form of the Glasgow composite measure pain scale (CMPS-SF), and visual analog scale (VAS) in dogs suffering from traumatic bone fractures before and after morphine administration in comparison with healthy dogs. METHODS: Fourteen dogs with hind limb or pelvic fractures and thirty healthy dogs were included. Dogs with fractures were divided into four groups according to analgesia received before participation. Physical examination, CMPS-SF, pain and stress behavior VAS scores were recorded in all dogs. Saliva and blood were collected once in healthy dogs and in dogs with fractures before and 35-70 min after morphine administration. Blood samples were analyzed for CST, VS, and cortisol. Saliva volumes, however, were insufficient for analysis. RESULTS: Catestatin and cortisol concentrations, and CMPS-SF, and VAS scores differed significantly between dogs with fractures prior to morphine administration and healthy dogs. After morphine administration, dogs with fractures had significantly decreased CMPS-SF and VAS scores and, compared to healthy dogs, CST concentrations, CMPS-SF, and VAS scores still differed significantly. However, CST concentrations remained largely within the normal range. Absolute delta values for CST significantly correlated with delta values for CMPS-SF. Catestatin and cortisol did not differ significantly before and after morphine administration. Vasostatin concentrations did not differ significantly between groups. CONCLUSIONS: Catestatin and cortisol concentrations, CMPS-SF, and VAS scores differed significantly in the dogs with traumatic bone fractures compared to the healthy dogs. Morphine treatment partially relieved pain and stress according to the subjective but not according to the objective assessments performed. However, because of the large degree of overlap with normal values, our results suggest that plasma CST concentrations have a limited potential as a clinically useful biomarker for pain-induced stress.

Efficacy of transsphenoidal hypophysectomy in treatment of dogs with pituitary-dependent hyperadrenocorticism.

The long-term survival, disease-free fractions, and the complications of hypophysectomy in 150 dogs with pituitary-dependent hyperadrenocorticism (PDH) were examined in a prospective study. Long-term survival and disease-free fractions in relation to pituitary size were analyzed by the Kaplan-Meijer estimate procedure. The 1-, 2-, 3-, and 4-year estimated survival rates were 84% (95% confidence interval [CI], 76-89%), 76% (67-83%), 72% (62-79%), and 68% (55-77%), respectively. Treatment failures included procedure-related mortalities (12 dogs) and incomplete hypophysectomies (9 dogs). The 1-, 2-, 3-, and 4-year estimated relapse-free fractions were 88% (CI: 80-93%), 75% (64-83%), 66% (54-76%), and 58% (45-70%), respectively. Postoperative reduction of tear production (58 eyes in 47 dogs) was often reversible but remained low until death in 11 eyes of 10 dogs. Central diabetes insipidus (CDI) occurred more frequently (62%) in dogs with enlarged pituitaries than in dogs with nonenlarged pituitaries (44%). Survival and disease-free fractions after hypophysectomy were markedly higher in dogs with nonenlarged pituitaries than in dogs with enlarged pituitaries. Transsphenoidal hypophysectomy is an effective treatment for PDH in dogs. The survival and disease-free fractions after hypophysectomy decrease and the incidence of CDI increases with increasing pituitary size. Therefore, early diagnosis of PDH is important and transsphenoidal hypophysectomy is expected to have the best outcome when used as primary treatment for dogs with nonenlarged or moderately enlarged pituitaries.

Stem cells in the canine pituitary gland and in pituitary adenomas.

Cushing's disease (CD) or pituitary-dependent hypercortisolism is a common endocrinopathy in dogs, with an estimated prevalence of 1 or 2 in 1000 dogs per year. It is caused by an adrenocorticotropic hormone secreting adenoma in the pars distalis or pars intermedia of the pituitary gland. The pituitary gland is a small endocrine gland located in the pituitary fossa. In the postnatal individual, the hypothalamus-pituitary axis plays a central role in maintaining homeostatic functions, like control of metabolism, reproduction, and growth. Stem cells are suggested to play a role in the homeostatic adaptations of the adult pituitary gland, such as the rapid specific cell-type expansion in response to pregnancy or lactation. Several cell populations have been suggested as pituitary stem cells, such as Side Population cells and cells expressing Sox2 or Nestin. These cell populations are discussed in this review. Also, stem and progenitor cells are thought to play a role in pituitary tumorigenesis, such as the development of pituitary adenomas in dogs. There are limited reports on the role of stem cells in pituitary adenomas, especially in dogs. Further studies are needed to identify and characterize this cell population and to develop specific cell targeting therapeutic strategies as a new way of treating canine CD.

Thorough investigation of a canine autoinflammatory disease (AID) confirms one main risk locus and suggests a modifier locus for amyloidosis.

Autoinflammatory disease (AID) manifests from the dysregulation of the innate immune system and is characterised by systemic and persistent inflammation. Clinical heterogeneity leads to patients presenting with one or a spectrum of phenotypic signs, leading to difficult diagnoses in the absence of a clear genetic cause. We used separate genome-wide SNP analyses to investigate five signs of AID (recurrent fever, arthritis, breed specific secondary dermatitis, otitis and systemic reactive amyloidosis) in a canine comparative model, the pure bred Chinese Shar-Pei. Analysis of 255 DNA samples revealed a shared locus on chromosome 13 spanning two peaks of association. A three-marker haplotype based on the most significant SNP (p<2.6×10(-8)) from each analysis showed that one haplotypic pair (H13-11) was present in the majority of AID individuals, implicating this as a shared risk factor for all phenotypes. We also noted that a genetic signature (F ST) distinguishing the phenotypic extremes of the breed specific Chinese Shar-Pei thick and wrinkled skin, flanked the chromosome 13 AID locus; suggesting that breed development and differentiation has played a parallel role in the genetics of breed fitness. Intriguingly, a potential modifier locus for amyloidosis was revealed on chromosome 14, and an investigation of candidate genes from both this and the chromosome 13 regions revealed significant (p<0.05) renal differential expression in four genes previously implicated in kidney or immune health (AOAH, ELMO1, HAS2 and IL6). These results illustrate that phenotypic heterogeneity need not be a reflection of genetic heterogeneity, and that genetic modifiers of disease could be masked if syndromes were not first considered as individual clinical signs and then as a sum of their component parts.

Role of Brg1 and HDAC2 in GR trans-repression of the pituitary POMC gene and misexpression in Cushing disease.

Negative feedback regulation of the proopiomelanocortin (POMC) gene by the glucocorticoid (Gc) receptor (GR) is a critical feature of the hypothalamo-pituitary-adrenal axis, and it is in part exerted by trans-repression between GR and the orphan nuclear receptors related to NGFI-B. We now show that Brg1, the ATPase subunit of the Swi/Snf complex, is essential for this trans-repression and that Brg1 is required in vivo to stabilize interactions between GR and NGFI-B as well as between GR and HDAC2. Whereas Brg1 is constitutively present at the POMC promoter, recruitment of GR and HDAC2 is ligand-dependent and results in histone H4 deacetylation of the POMC locus. In addition, GR-dependent repression inhibits promoter clearance by RNA polymerase II. Thus, corecruitment of repressor and activator at the promoter and chromatin modification jointly contribute to trans-repression initiated by direct interactions between GR and NGFI-B. Loss of Brg1 or HDAC2 should therefore produce Gc resistance, and we show that approximately 50% of Gc-resistant human and dog corticotroph adenomas, which are the hallmark of Cushing disease, are deficient in nuclear expression of either protein. In addition to providing a molecular basis for Gc resistance, these deficiencies may also contribute to the tumorigenic process.