CRISPR/Cas9 generated knockout mice lacking phenylalanine hydroxylase protein as a novel preclinical model for human phenylketonuria.

Phenylketonuria (PKU) is an autosomal recessive inborn error of L-phenylalanine (Phe) metabolism. It is caused by a partial or complete deficiency of the enzyme phenylalanine hydroxylase (PAH), which is necessary for conversion of Phe to tyrosine (Tyr). This metabolic error results in buildup of Phe and reduction of Tyr concentration in blood and in the brain, leading to neurological disease and intellectual deficits. Patients exhibit retarded body growth, hypopigmentation, hypocholesterolemia and low levels of neurotransmitters. Here we report first attempt at creating a homozygous Pah knock-out (KO) (Hom) mouse model, which was developed in the C57BL/6 J strain using CRISPR/Cas9 where codon 7 (GAG) in Pah gene was changed to a stop codon TAG. We investigated 2 to 6-month-old, male, Hom mice using comprehensive behavioral and biochemical assays, MRI and histopathology. Age and sex-matched heterozygous Pah-KO (Het) mice were used as control mice, as they exhibit enough PAH enzyme activity to provide Phe and Tyr levels comparable to the wild-type mice. Overall, our findings demonstrate that 6-month-old, male Hom mice completely lack PAH enzyme, exhibit significantly higher blood and brain Phe levels, lower levels of brain Tyr and neurotransmitters along with lower myelin content and have significant behavioral deficit. These mice exhibit phenotypes that closely resemble PKU patients such as retarded body growth, cutaneous hypopigmentation, and hypocholesterolemia when compared to the age- and sex-matched Het mice. Altogether, biochemical, behavioral, and pathologic features of this novel mouse model suggest that it can be used as a reliable translational tool for PKU preclinical research and drug development.

Obesity Increases Prevalence of Colonic Adenomas at Screening Colonoscopy: A Canadian Community-Based Study.

BACKGROUND AND AIMS: Obesity is a risk factor for colorectal neoplasia. We examined the influence of obesity and metabolic syndrome (MetS) on prevalence of neoplasia at screening colonoscopy. METHODS: We evaluated 2020 subjects undergoing first screening colonoscopy. Body mass index (BMI) was calculated at enrolment. Hyperlipidemia (HL), hypertension (HT), and diabetes mellitus (DM) were identified. Details of colonoscopy, polypectomy, and histology were recorded. Odds for adenomas (A) and advanced adenomas (ADV) in overweight (BMI 25.1-30) and obese (BMI > 30) subjects were assessed by multinomial regression, adjusted for covariates. Analyses included relationships between HL, HT, DM, age, tobacco usage, and neoplasia. Discriminatory power of HT, HL, DM, and BMI for neoplasia was assessed by binary logistic regression. Odds were calculated for neoplasia in each colonic segment related to BMI. RESULTS: A and ADV were commoner in overweight and obese males, obese females, older subjects, and smokers. HL, HT, and DM were associated with increased odds for neoplasia, significantly for A with hypertension. BMI alone predicted neoplasia as well as HT, HL, DM, or combinations thereof. All segments of the colon were affected. Multiple polyps were particularly prevalent in the obese. CONCLUSIONS: Obesity and MetS are risk factors for colonic neoplasia in a Canadian population.