Therapeutic homology-independent targeted integration in retina and liver

Challenges to the widespread application of gene therapy with adeno-associated viral (AAV) vectors include dominant conditions due to gain-of-function mutations which require allele-specific knockout, as well as long-term transgene expression from proliferating tissues, which is hampered by AAV DNA episomal status. To overcome these challenges, we used CRISPR/Cas9-mediated homology-independent targeted integration (HITI) in retina and liver as paradigmatic target tissues. We show that AAV-HITI targets photoreceptors of both mouse and pig retina, and this results in significant improvements to retinal morphology and function in mice with autosomal dominant retinitis pigmentosa. In addition, we show that neonatal systemic AAV-HITI delivery achieves stable liver transgene expression and phenotypic improvement in a mouse model of a severe lysosomal storage disease. We also show that HITI applications predominantly result in on-target editing. These results lay the groundwork for the application of AAV-HITI for the treatment of diseases affecting various organs

Exercise-induced rhabdomyolysis and transient loss of deambulation as outset of partial carnitine palmityl transferase II deficiency.

We report the case of a 13-year-old boy with an abrupt onset of leg pain and muscle weakness, incapability of deambulation and a laboratory picture of exercise-induced acute rhabdomyolysis. Intravenous hyperhydration and forced diuresis were adopted to avoid renal complications. No evidence of articular or residual muscular damage was appreciated in the short-term. The recurrence of rhabdomyolysis required a muscular biopsy showing a disturbance of fatty acid beta-oxidation pathway

Details of cytoplasm-membranous CD56 positivity in a benign lesion diagnosed as BN (avidin-biotin-peroxidase complex x400);

<p>b) Details of diffuse cytoplasm and membrane CD56 expression on the histological sample for the same case (avidin-biotin-peroxidase complex x300).</p

Details of a thyroid lesion diagnosed as positive for malignancy favoring papillary thyroid carcinoma on LBC, in the upper left a detail of nuclear inclusion (PAPx400).

<p>Details of a thyroid lesion diagnosed as positive for malignancy favoring papillary thyroid carcinoma on LBC, in the upper left a detail of nuclear inclusion (PAPx400).</p

Clinicopathologic features of 85 thyroid lesions with BN and PM diagnoses and histological follow-up.

<p>BN: Benign Nodule; PM: Positive of Malignancy favoring Papillary Thyroid cancer; M/: male/female; Lfn Mets: Lymph nodes metastases; PTC: papillary thyroid carcinoma; FVPC: follicular variant of PTC; TCV: Tall cell variant of PTC; °FVPCs included 11encapsulated and 10 infiltrative variants.</p><p>Clinicopathologic features of 85 thyroid lesions with BN and PM diagnoses and histological follow-up.</p

Descriptive statistics for immunocytochemical markers used in double and triple combinations in the concordant cyto-histological series^§.

<p>Sens: Sensitivity; Spec: Specificity; Diagn. Acc.: Diagnostic accuracy; OR (95% CI): Odds ratio (95% Confidence intervals). A p-value less than 0.05 has been considered significant.</p><p>^§Data for the discordant ICC cases were reported in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0132939#pone.0132939.s001" target="_blank">S1 Table</a></p><p>Descriptive statistics for immunocytochemical markers used in double and triple combinations in the concordant cyto-histological series<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0132939#t005fn002" target="_blank">^§</a>.</p

Details of cytoplasm-membranous CD56 positivity in a benign lesion diagnosed as BN (avidin-biotin-peroxidase complex x400);

<p>b) Details of diffuse cytoplasm and membrane CD56 expression on the histological sample for the same case (avidin-biotin-peroxidase complex x300).</p

Descriptive statistics for each immunocytochemical marker in the cyto-histological series.

<p>Sens: Sensitivity; Spec: Specificity; Diagn. Acc.: Diagnostic accuracy; OR (95% CI): Odds ratio (95% Confidence intervals). A p-value less than 0.05 is considered significant.</p><p>Descriptive statistics for each immunocytochemical marker in the cyto-histological series.</p