Synthetic AAV/CRISPR vectors for blocking HIV-1 expression in persistently infected astrocytes.

Astrocytes, the most abundant cells in the mammalian brain, perform key functions and are involved in several neurodegenerative diseases. The human immunodeficiency virus (HIV) can persist in astrocytes, contributing to the HIV burden and neurological dysfunctions in infected individuals. While a comprehensive approach to HIV cure must include the targeting of HIV-1 in astrocytes, dedicated tools for this purpose are still lacking. Here we report a novel Adeno-associated virus-based vector (AAV9P1) with a synthetic surface peptide for transduction of astrocytes. Analysis of AAV9P1 transduction efficiencies with single brain cell populations, including primary human brain cells, as well as human brain organoids demonstrated that AAV9P1 targeted terminally differentiated human astrocytes much more efficiently than neurons. We then investigated whether AAV9P1 can be used to deliver HIV-inhibitory genes to astrocytes. To this end we generated AAV9P1 vectors containing genes for HIV-1 proviral editing by CRISPR/Cas9. Latently HIV-1 infected astrocytes transduced with these vectors showed significantly diminished reactivation of proviruses, compared with untransduced cultures. Sequence analysis identified mutations/deletions in key HIV-1 transcriptional control regions. We conclude that AAV9P1 is a promising tool for gene delivery to astrocytes and may facilitate inactivation/destruction of persisting HIV-1 proviruses in astrocyte reservoirs.

T cells with low CD2 levels express reduced restriction factors and are preferentially infected in therapy naïve chronic HIV-1 patients.

INTRODUCTION: Restriction factors (RFs) suppress HIV-1 in cell lines and primary cell models. Hence, RFs might be attractive targets for novel antiviral strategies, but their importance for virus control in vivo is controversial. METHODS: We profiled the expression of RFs in primary blood-derived mononuclear cells (PBMC) from therapy-naïve HIV-1 patients and quantified infection. RESULTS: Overall, there was no correlation between individual RF expression and HIV-1 status in total PBMC. However, we identified a T cell population with low levels of intracellular CD2 and reduced expression of SAMHD1, p21 and SerinC5. CD2low T cells with reduced RF expression were markedly positive for HIV-1 p24. In contrast, CD2+ T cells were less infected and expressed higher levels of RFs. CD2low T cell infection correlated with viral loads and was associated with HIV-1 disease progression. CONCLUSIONS: In untreated therapy naïve chronic HIV-1 patients, RF expression in T cells is associated with CD2 expression and seems to influence viral loads. Our study suggests that RFs help to control HIV-1 infection in certain T cells in vivo and supports the potential for RFs as promising targets for therapeutic intervention.

Diffuse idiopathic skeletal hyperostosis (DISH) of the elbow: a controlled radiological study.

BACKGROUND: Extraspinal manifestations of diffuse idiopathic skeletal hyperostosis (DISH) have been described previously. We aimed to assess the prevalence of elbow hyperostotic spurs, to search for sites discriminating for elbow DISH and to analyze the effect of physical activities, handedness and sex. METHODS: Out of 284 patients hospitalized for extraskeletal disorders, 85 patients (33 with and 52 without thoracospinal DISH) agreed to bilateral elbow X-rays in two projections. Clinical information was collected by a standardized questionnaire and X-rays were graded blindly. RESULTS: A total of 400 hyperostotic spurs (210 unilateral, 95 bilateral) were present at 11 predefined sites. The most frequent sites affected were the olecranon (20.8%), lateral epicondyle (17.8%) and medial epicondyle (15.5%). In carriers of thoracospinal DISH significantly more hyperostotic spurs were present at the lateral and medial epicondyle compared to non-DISH carriers (OR 4.01 [95% CI 1.35-12.34] and 2.88 [1.03-8.24], respectively). The olecranon, lateral and medial epicondyle contributed significantly to the classification of elbow DISH (OR 22.2 [4.1-144.7], 9.6 [1.9-61.2] and 10.1 [2.2-52.1], respectively). The prevalence of elbow hyperostotic spurs was higher in 45 patients with a history of heavy physical activities (24.4% versus 18.0%, OR 1.48 [1.17-1.86]), at the right elbow (24.2% versus 18.6%, OR 1.39 [1.11-1.75]) and in 62 males (22.8% versus 17.6%, OR 1.38 [1.06-1.81]). CONCLUSIONS: Hyperostotic spurs at the olecranon, lateral and medial epicondyle had the highest prevalence and disclosed the most pronounced discrimination for elbow DISH. Mechanical factors such as physical activities and handedness, and sex influenced the formation of these spurs.