High-Level Expression of Alkaline Phosphatase by Adeno-Associated Virus Vector Ameliorates Pathological Bone Structure in a Hypophosphatasia Mouse Model.

Hypophosphatasia (HPP) is a systemic skeletal disease caused by mutations in the gene encoding tissue-nonspecific alkaline phosphatase (TNALP). We recently reported that survival of HPP model mice can be prolonged using an adeno-associated virus (AAV) vector expressing bone-targeted TNALP with deca-aspartate at the C terminus (TNALP-D10); however, abnormal bone structure and hypomineralization remained in the treated mice. Here, to develop a more effective and clinically applicable approach, we assessed whether transfection with TNALP-D10 expressing virus vector at a higher dose than previously used would ameliorate bone structure defects. We constructed a self-complementary AAV8 vector expressing TNALP driven by the chicken beta-actin (CBA) promoter (scAAV8-CB-TNALP-D10). The vector was injected into both quadriceps femoris muscles of newborn HPP mice at a dose of 4.5 × 1012 vector genome (v.g.)/body, resulting in 20 U/mL of serum ALP activity. The 4.5 × 1012 v.g./body-treated HPP mice grew normally and displayed improved bone structure at the knee joints in X-ray images. Micro-CT analysis showed normal trabecular bone structure and mineralization. The mechanical properties of the femur were also recovered. Histological analysis of the femurs demonstrated that ALP replacement levels were sufficient to promote normal, growth plate cartilage arrangement. These results suggest that AAV vector-mediated high-dose TNALP-D10 therapy is a promising option for improving the quality of life (QOL) of patients with the infantile form of HPP.

Incidence of canine glaucoma with goniodysplasia in Japan : a retrospective study.

The incidence of primary and secondary glaucoma in dogs was investigated. A total of 1244 dogs received ophthalmologic examinations, including tonometry and gonioscopy. Goniophotographs were taken using a goniolens to evaluate the iridocorneal angle (ICA) as well as pectinate ligament (PL). The anterior width of the ciliary cleft and the total distance from the origin of the PL to the anterior corneal surface were measured from the goniophotographs. Glaucoma was diagnosed based on the cupping of the optic nerve head, clinical signs, ocular changes, and high IOP, and it was synchronized with gonioscopic grades to differentiate between primary and secondary glaucoma. We investigated 1244 dogs of 29 breeds, including the mixed breed; among these, glaucoma was diagnosed in 127 dogs (162 eyes). Of 162 eyes, primary glaucoma was diagnosed in 129 eyes and secondary glaucoma in 33 eyes. Shiba Inu dogs (42 dogs, 33%) showed the highest incidence of glaucoma, followed by Shih-Tzu (21 dogs, 16.5%). Furthermore, all the glaucomatous Shiba Inu dogs had primary glaucoma with abnormal ICA grades and dysplastic PLs. The findings of our study reveal that the Shiba Inu breed in Japan may have a hereditary predisposition to glaucoma.