Isolation of Next-Generation Gene Therapy Vectors through Engineering, Barcoding, and Screening of Adeno-Associated Virus (AAV) Capsid Variants.

Gene delivery vectors derived from Adeno-associated virus (AAV) are one of the most promising tools for the treatment of genetic diseases, evidenced by encouraging clinical data and the approval of several AAV gene therapies. Two major reasons for the success of AAV vectors are (i) the prior isolation of various naturally occurring viral serotypes with distinct properties, and (ii) the subsequent establishment of powerful technologies for their molecular engineering and repurposing in high throughput. Further boosting the potential of these techniques are recently implemented strategies for barcoding selected AAV capsids on the DNA and RNA level, permitting their comprehensive and parallel in vivo stratification in all major organs and cell types in a single animal.

Boosters for adeno-associated virus (AAV) vector (r)evolution.

Adeno-associated virus (AAV) is one of the most exciting and most versatile templates for engineering of gene-delivery vectors for use in human gene therapy, owing to the existence of numerous naturally occurring capsid variants and their amenability to directed molecular evolution. As a result, the field has witnessed an explosion of novel "designer" AAV capsids and ensuing vectors over the last two decades, which have been isolated from comprehensive capsid libraries generated through technologies such as DNA shuffling or peptide display, and stratified under stringent positive and/or negative selection pressures. Here, we briefly highlight a panel of recent, innovative and transformative methodologies that we consider to have exceptional potential to advance directed AAV capsid evolution and to thereby accelerate AAV vector revolution.

Auto-Regulation of the Thyroid Gland Beyond Classical Pathways.

This mini-review asks how self-regulation of the thyroid gland is realized at the cellular and molecular levels by canonical and non-canonical means. Canonical pathways of thyroid regulation comprise thyroid stimulating hormone-triggered receptor signaling. As part of non-canonical regulation, we hypothesized an interplay between protease-mediated thyroglobulin processing and thyroid hormone release into the circulation by means of thyroid hormone transporters like Mct8.

Treatment of rat thyrocytes in vitro with cathepsin B and L inhibitors results in disruption of primary cilia leading to redistribution of the trace amine associated receptor 1 to the endoplasmic reticulum.

Taar1 is a G protein-coupled receptor (GPCR) confined to primary cilia of rodent thyroid epithelial cells. Taar1-deficient mouse thyroid follicles feature luminal accumulation of thyroglobulin suggesting that Taar1 acts as a regulator of extra- and pericellular thyroglobulin processing, which is mediated by cysteine cathepsin proteases present at the apical plasma membrane of rodent thyrocytes. Here, by immunostaining and confocal laser scanning microscopy, we demonstrated co-localization of cathepsin L, but only little cathepsin B, with Taar1 at primary cilia of rat thyrocytes, the FRT cells.

Canonical TSH Regulation of Cathepsin-Mediated Thyroglobulin Processing in the Thyroid Gland of Male Mice Requires Taar1 Expression.

Trace amine-associated receptor 1 (Taar1) has been suggested as putative receptor of thyronamines. These are aminergic messengers with potential metabolic and neurological effects countering their contingent precursors, the thyroid hormones (THs). Recently, we found Taar1 to be localized at the primary cilia of rodent thyroid epithelial cells and .

Trace Amine-Associated Receptor 1 Localization at the Apical Plasma Membrane Domain of Fisher Rat Thyroid Epithelial Cells Is Confined to Cilia.

Background: The trace amine-associated receptor 1 (Taar1) is one member of the Taar family of G-protein-coupled receptors (GPCR) accepting various biogenic amines as ligands. It has been proposed that Taar1 mediates rapid, membrane-initiated effects of thyronamines, the endogenous decarboxylated and deiodinated relatives of the classical thyroid hormones T4 and T3.

Objectives: Although the physiological actions of thyronamines in general and 3-iodothyronamine (T1AM) in particular are incompletely understood, studies published to date suggest that synthetic T1AM-activated Taar1 signaling antagonizes thyromimetic effects exerted by T3.

Expression of pre-selected TMEMs with predicted ER localization as potential classifiers of ccRCC tumors.

Background: VHL inactivation is the most established molecular characteristic of clear cell renal cell carcinoma (ccRCC), with only a few additional genes implicated in development of this kidney tumor. In recently published ccRCC gene expression meta-analysis study we identified a number of deregulated genes with limited information available concerning their biological role, represented by gene transcripts belonging to transmembrane proteins family (TMEMs). TMEMs are predicted to be components of cellular membranes, such as mitochondrial membranes, ER, lysosomes and Golgi apparatus.

[Cerebral functional asymmetry in face perception by the patients with focal brain damage].

The results are reported of the study of face perception in a group of 18 healthy subjects and 10 patients with lesions in the right or left cerebral hemisphere. The task given to these subjects included identification of a face photograph exposed for 20 msec in the right or left field of vision. The index of perception coefficient was the number of errors made while addressing the material to the left or right cerebral hemisphere.

[The process of learning verbal material in patients with various damaged cerebral structures].

The results are described of a study of the process of verbal material learning and its recall after distraction in 20 patients with involuntary movements treated by thalamotomy, and in 20 epileptic patients treated by amygdalotomy and hippocampotomy. In both groups learning difficulties had been noted already before the operation. In postoperative tests these difficulties were found to have increased immediately after the surgical intervention in patients with lesions in the left cerebral hemisphere.

Cerebral hemispheric differences in visual perception of various types of verbal material.

Hemispheric differences in perception of various verbal material were sought in two groups of Ss: 10 normal adults (a control group) and 10 patients of the Neurosurgery Clinic with focal brain damage either to the left or right hemisphere. The stimuli were letters projected singly, in three-letter nonsense sequences or in three letter Polish words on a screen in the left or right visual half-field. The results obtained from the control groups indicate that the dominance of the left hemisphere is limited only to the meaningful verbal material.

[Use of haemodilution in the treatment of cranio-cerebral injuries (author's transl)].

By decreasing blood viscosity and vascular resistance normovolemic haemodilution increases cerebral blood flow and improves microcirculation. This eliminates the symptoms of anoxia of cerebral tissue. The extent of disturbances of the blood brain barrier is decreased.