Base and Prime Editing in the Retina-From Preclinical Research toward Human Clinical Trials.

Inherited retinal diseases (IRDs) are a clinically and genetically heterogeneous group of diseases that are one of the leading causes of vision loss in young and aged individuals. IRDs are mainly caused by a loss of the post-mitotic photoreceptor neurons of the retina, or by the degeneration of the retinal pigment epithelium. Unfortunately, once these cells are damaged, it is irreversible and leads to permanent vision impairment. Thought to be previously incurable, gene therapy has been rapidly evolving to be a potential treatment to prevent further degeneration of the retina and preserve visual function. The development of clustered regularly interspaced short palindromic repeats (CRISPR)-CRISPR-associated protein 9 (Cas9) base and prime editors have increased the capabilities of the genome editing toolbox in recent years. Both base and prime editors evade the creation of double-stranded breaks in deoxyribonucleic acid (DNA) and the requirement of donor template of DNA for repair, which make them advantageous methods in developing clinical therapies. In addition, establishing a permanent edit within the genome could be better suited for patients with progressive degeneration. In this review, we will summarize published uses of successful base and prime editing in treating IRDs.

Single-nucleotide-level mapping of DNA regulatory elements that control fetal hemoglobin expression.

Pinpointing functional noncoding DNA sequences and defining their contributions to health-related traits is a major challenge for modern genetics. We developed a high-throughput framework to map noncoding DNA functions with single-nucleotide resolution in four loci that control erythroid fetal hemoglobin (HbF) expression, a genetically determined trait that modifies sickle cell disease (SCD) phenotypes. Specifically, we used the adenine base editor ABEmax to introduce 10,156 separate A•T to G•C conversions in 307 predicted regulatory elements and quantified the effects on erythroid HbF expression. We identified numerous regulatory elements, defined their epigenomic structures and linked them to low-frequency variants associated with HbF expression in an SCD cohort. Targeting a newly discovered γ-globin gene repressor element in SCD donor CD34+ hematopoietic progenitors raised HbF levels in the erythroid progeny, inhibiting hypoxia-induced sickling. Our findings reveal previously unappreciated genetic complexities of HbF regulation and provide potentially therapeutic insights into SCD.

Dynamic CTCF binding directly mediates interactions among cis-regulatory elements essential for hematopoiesis.

While constitutive CCCTC-binding factor (CTCF)-binding sites are needed to maintain relatively invariant chromatin structures, such as topologically associating domains, the precise roles of CTCF to control cell-type-specific transcriptional regulation remain poorly explored. We examined CTCF occupancy in different types of primary blood cells derived from the same donor to elucidate a new role for CTCF in gene regulation during blood cell development. We identified dynamic, cell-type-specific binding sites for CTCF that colocalize with lineage-specific transcription factors. These dynamic sites are enriched for single-nucleotide polymorphisms that are associated with blood cell traits in different linages, and they coincide with the key regulatory elements governing hematopoiesis. CRISPR-Cas9-based perturbation experiments demonstrated that these dynamic CTCF-binding sites play a critical role in red blood cell development. Furthermore, precise deletion of CTCF-binding motifs in dynamic sites abolished interactions of erythroid genes, such as RBM38, with their associated enhancers and led to abnormal erythropoiesis. These results suggest a novel, cell-type-specific function for CTCF in which it may serve to facilitate interaction of distal regulatory emblements with target promoters. Our study of the dynamic, cell-type-specific binding and function of CTCF provides new insights into transcriptional regulation during hematopoiesis.

Spatial Clustering of Dengue Fever Incidence and Its Association with Surrounding Greenness.

With more than 58,000 cases reported by the country's Centers for Disease Control, the dengue outbreaks from 2014 to 2015 seriously impacted the southern part of Taiwan. This study aims to assess the spatial autocorrelation of the dengue fever (DF) outbreak in southern Taiwan in 2014 and 2015, and to further understand the effects of green space (such as forests, farms, grass, and parks) allocation on DF. In this study, two different greenness indexes were used. The first green metric, the normalized difference vegetation index (NDVI), was provided by the long-term NASA MODIS satellite NDVI database, which quantifies and represents the overall vegetation greenness. The latest 2013 land use survey GIS database completed by the National Land Surveying and Mapping Center was obtained to access another green metric, green land use in Taiwan. We first used Spearman's rho to find out the relationship between DF and green space, and then three spatial autocorrelation methods, including Global Moran's I, high/low clustering, and Hot Spot were employed to assess the spatial autocorrelation of DF outbreak. In considering the impact of social and environmental factors in DF, we used generalized linear mixed models (GLMM) to further clarify the relationship between different types of green land use and dengue cases. Results of spatial autocorrelation analysis showed a high aggregation of dengue epidemic in southern Taiwan, and the metropolitan areas were the main hotspots. Results of correlation analysis and GLMM showed a positive correlation between parks and dengue fever, and the other five green space metrics and land types revealed a negative association with DF. Our findings may be an important asset for improving surveillance and control interventions for dengue.

Metabolic and toxicological considerations of newly approved prostate cancer drugs.

INTRODUCTION: Despite increasing early detection and treatment of prostate cancer, a subset of patients presents with or develops metastatic disease. Androgen deprivation therapy is effective but resistance eventually develops, resulting in a lethal phenotype known as castration-resistant prostate cancer (CRPC). Recently, several novel treatments, each with distinct mechanisms of actions, have been approved for the treatment of CRPC. Understanding of the metabolic and toxicological considerations of each treatment is crucial to the successful management of patients with this lethal disease. AREAS COVERED: The present review focuses on the metabolism and toxicology characteristics of recently approved therapies in the treatment of metastatic CRPC. Specifically, the authors review the mechanism of action of these therapies in addition to their, efficacy and usage recommendations for hepatic and renal impairment. The authors, furthermore, also consider their adverse effect profile. EXPERT OPINION: Despite the expanding armamentarium of effective treatments for CRPC, the exact choice, timing and sequence of various therapies remains an inexact science and requires further investigation. Variations in patient comorbidities, disease burden, organ functions and adverse events are all critical determinants in selection of treatment. Identification and validation of molecular pathways and specific targets that drives disease progression will be critical in the continued development of effective treatments in advanced prostate cancer.

Targeting the androgen receptor signalling axis in castration-resistant prostate cancer (CRPC).

What's known on the subject? and What does the study add? Castration resistance has been appreciated for decades, and several mechanisms theorising on this effect have been proposed. A rich pipeline of novel agents, including abiraterone and MDV3100, have provided proof of principle that novel agents targeting the AR signalling pathway with superior selectivity and activity than predecessors have yielded significant clinical benefit for patients with metastatic castration-resistant prostate cancer. Our review provides an update in the development of several novel agents targeting the AR signalling pathway now in clinical testing, as well as review novel therapies in development with distinct mechanisms of action showing promising preclinical activity. • Despite undergoing local therapy with curative intent, 20-30% of patients with prostate cancer will ultimately development metastatic disease, leading to morbidity and mortality. • Androgen-deprivation therapy (ADT) for men with metastatic prostate cancer results in transient clinical benefit, but ultimately, cancers progress despite castrate levels of serum testosterone, a clinical state classically referred to as 'hormone refractory' disease. • In this review, we examine mechanisms of resistance to ADT that have redefined our understanding of the more appropriately termed 'castration resistant' disease, and have paved the way for a new generation of therapeutics targeting the androgen signalling axis in advanced prostate cancer.