Targeting miR-181a/b in retinitis pigmentosa: implications for disease progression and therapy.

BACKGROUND: Retinitis pigmentosa (RP) is a genetically heterogeneous group of degenerative disorders causing progressive vision loss due to photoreceptor death. RP affects other retinal cells, including the retinal pigment epithelium (RPE). MicroRNAs (miRs) are implicated in RP pathogenesis, and downregulating miR-181a/b has shown therapeutic benefit in RP mouse models by improving mitochondrial function. This study investigates the expression profile of miR-181a/b in RPE cells and the neural retina during RP disease progression. We also evaluate how miR-181a/b downregulation, by knocking out miR-181a/b-1 cluster in RPE cells, confers therapeutic efficacy in an RP mouse model and explore the mechanisms underlying this process. RESULTS: Our findings reveal distinct expression profiles, with downregulated miR-181a/b in RPE cells suggesting a protective response and upregulated miR-181a/b in the neural retina indicating a role in disease progression. We found that miR-181a/b-2, encoded in a separate genomic cluster, compensates for miR-181a/b-1 ablation in RPE cells at late time points. The transient downregulation of miR-181a/b in RPE cells at post-natal week 6 (PW6) led to improved RPE morphology, retarded photoreceptor degeneration and decreased RPE aerobic glycolysis. CONCLUSIONS: Our study elucidates the underlying mechanisms associated with the therapeutic modulation of miR-181a/b, providing insights into the metabolic processes linked to its RPE-specific downregulation. Our data further highlights the impact of compensatory regulation between miR clusters with implications for the development of miR-based therapeutics.

Risk Factors of Mortality and Long-Term Survival in Burn Patients With/Without Extracorporeal Membrane Oxygenation: A 16 Year Real-World Study.

Burn patients face cardiopulmonary failure risks, with recent observational studies suggesting promising outcomes for extracorporeal membrane oxygenation (ECMO). However, the effectiveness and long-term survival remain unclear. Our study aims to assess mortality risk factors and long-term survival in burn patients with and without ECMO. This study used Taiwan's National Health Insurance Research Database and designed a case-control with onefold propensity score matching across variables including sex, age, total body surface area (TBSA) burned, and index date. We analyzed mortality and survival risk factors in each stratified group with/without ECMO. Finally, we analyze the mortality according to ECMO and TBSA burned, and the cause of death and long-term survival. From 2000 to 2015, 4,556 burn patients with ECMO compared to an equivalent number without ECMO. Primary mortality include male, age >65, TBSA ≥30%, escharotomy, hemodialysis, and bacteremia. The ECMO group showed lower survival across all stratified risk factors, with the primary cause of death being burn-related issues, followed by respiratory and heart failure. The overall mortality rate was 54.41% with ECMO and 40.94% without ECMO (p < 0.001). Additionally, long-term survival is lower in the group with ECMO. This research provides a valuable real-world gross report about ECMO efficacy and long-term survival among burn patients with/without ECMO.

A parametric bootstrap approach for computing confidence intervals for genetic correlations with application to genetically determined protein-protein networks.

Genetic correlation refers to the correlation between genetic determinants of a pair of traits. When using individual-level data, it is typically estimated based on a bivariate model specification where the correlation between the two variables is identifiable and can be estimated from a covariance model that incorporates the genetic relationship between individuals, e.g., using a pre-specified kinship matrix. Inference relying on asymptotic normality of the genetic correlation parameter estimates may be inaccurate when the sample size is low, when the genetic correlation is close to the boundary of the parameter space, and when the heritability of at least one of the traits is low. We address this problem by developing a parametric bootstrap procedure to construct confidence intervals for genetic correlation estimates. The procedure simulates paired traits under a range of heritability and genetic correlation parameters, and it uses the population structure encapsulated by the kinship matrix. Heritabilities and genetic correlations are estimated using the close-form, method of moment, Haseman-Elston regression estimators. The proposed parametric bootstrap procedure is especially useful when genetic correlations are computed on pairs of thousands of traits measured on the same exact set of individuals. We demonstrate the parametric bootstrap approach on a proteomics dataset from the Jackson Heart Study.

Defect Engineering Strategy for Superior Integration of Metal-Organic Framework and Halide Perovskite as a Fluorescence Sensing Material.

Combining halide perovskite quantum dots (QDs) and metal-organic frameworks (MOFs) material is challenging when the QDs' size is larger than the MOFs' nanopores. Here, we adopted a simple defect engineering approach to increase the size of zeolitic imidazolate framework 90 (ZIF-90)'s pores size to better load CH3NH3PbBr3 perovskite QDs. This defect structure effect can be easily achieved by adjusting the metal-to-ligand ratio throughout the ZIF-90 synthesis process. The QDs are then grown in the defective structure, resulting in a hybrid ZIF-90-perovskite (ZP) composite. The QDs in ZP composites occupied the gap of 10-18 nm defective ZIF-90 crystal and interestingly isolated the QDs with high stability in aqueous solution. We also investigated the relationship between defect engineering and fluorescence sensing, finding that the aqueous Cu2+ ion concentration was directly correlated to defective ZIF-90 and ZP composites. We also found that the role of the O-Cu coordination bonds and CH3NHCu+ species formation in the materials when they reacted with Cu2+ was responsible for this relationship. Finally, this strategy was successful in developing Cu2+ ion fluorescence sensing in water with better selectivity and sensitivity.

Porphyromonas gingivalis GroEL exacerbates orthotopic allograft transplantation vasculopathy via impairment of endothelial cell function.

Orthotopic allograft transplantation (OAT) is a significant approach to addressing organ failure. However, persistent immune responses to the allograft affect chronic rejection, which induces OAT vasculopathy (OATV) and organ failure. Porphyromonas gingivalis can infiltrate remote organs via the bloodstream, thereby intensifying the severity of cardiovascular, respiratory, and neurodegenerative diseases and cancer. GroEL, a virulence factor of P. gingivalis promotes pro-inflammatory cytokine production in host cells, which assumes to play a pivotal role in the pathogenesis of cardiovascular diseases. Although the aggravation of OATV is attributable to numerous factors, the role of GroEL remains ambiguous. Therefore, this study aimed to investigate the impact of GroEL on OATV. Aortic grafts extracted from PVG/Seac rats were transplanted into ACI/NKyo rats and in vitro human endothelial progenitor cell (EPC) and coronary artery endothelial cell (HCAEC) models. The experimental findings revealed that GroEL exacerbates OATV in ACI/NKyo rats by affecting EPC and smooth muscle progenitor cell (SMPC) function and enabling the anomalous accumulation of collagen. In vitro, GroEL spurs endothelial-mesenchymal transition in EPCs, reduces HCAEC tube formation and barrier function by downregulating junction proteins, accelerates HCAEC aging by lowering mitochondrial membrane potential and respiratory function, and impedes HCAEC migration by modulating cytoskeleton-associated molecules. This study suggests that P. gingivalis GroEL could potentially augment OATV by impacting vascular progenitor and endothelial cell functions.

Targeted therapy in glomerular diseases.

Targeted therapy has emerged as a more precise approach to treat glomerular diseases, focusing on specific molecular or cellular processes that contribute to disease development or progression. This approach complements or replaces traditional immunosuppressive therapy, optimizes supportive care, and provides a more personalized treatment strategy. In this review, we summarize the evolving understanding of pathogenic mechanisms in immune-mediated glomerular diseases and the developing targeted therapies based on these mechanisms. We begin by discussing pan-B-cell depletion, anti-CD20 rituximab, and targeting B-cell survival signaling through the BAFF/APRIL pathway. We also exam specific plasma cell depletion with anti-CD38 antibody. We then shift our focus to complement activation in glomerular diseases, which is involved in antibody-mediated glomerular diseases, such as IgA nephropathy, membranous nephropathy, ANCA-associated vasculitis, and lupus nephritis. Non-antibody-mediated complement activation occurs in glomerular diseases, including C3 glomerulopathy, complement-mediated atypical hemolytic uremic syndrome, and focal segmental glomerulosclerosis. We discuss specific inhibition of terminal, lectin, and alternative pathways in different glomerular diseases. Finally, we summarize current clinical trials targeting the final pathways of various glomerular diseases, including kidney fibrosis. We conclude that targeted therapy based on individualized pathogenesis should be the future of treating glomerular diseases.

GroEL of Porphyromonas gingivalis-induced microRNAs accelerate tumor neovascularization by downregulating thrombomodulin expression in endothelial progenitor cells.

We found that GroEL in Porphyromonas gingivalis accelerated tumor growth and increased mortality in tumor-bearing mice; GroEL promoted proangiogenic function, which may be the reason for promoting tumor growth. To understand the regulatory mechanisms by which GroEL increases the proangiogenic function of endothelial progenitor cells (EPCs), we explored in this study. In EPCs, MTT assay, wound-healing assay, and tube formation assay were performed to analyze its activity. Western blot and immunoprecipitation were used to study the protein expression along with next-generation sequencing for miRNA expression. Finally, a murine tumorigenesis animal model was used to confirm the results of in vitro. The results indicated that thrombomodulin (TM) direct interacts with PI3 K/Akt to inhibit the activation of signaling pathways. When the expression of TM is decreased by GroEL stimulation, molecules in the PI3 K/Akt signaling axis are released and activated, resulting in increased migration and tube formation of EPCs. In addition, GroEL inhibits TM mRNA expression by activating miR-1248, miR-1291, and miR-5701. Losing the functions of miR-1248, miR-1291, and miR-5701 can effectively alleviate the GroEL-induced decrease in TM protein levels and inhibit the proangiogenic abilities of EPCs. These results were also confirmed in animal experiments. In conclusion, the intracellular domain of the TM of EPCs plays a negative regulatory role in the proangiogenic capabilities of EPCs, mainly through direct interaction between TM and PI3 K/Akt to inhibit the activation of signaling pathways. The effects of GroEL on tumor growth can be reduced by inhibiting the proangiogenic properties of EPCs through the inhibition of the expression of specific miRNAs.

Advancing Near-Infrared Light Sources: Enhancing Chromium Emission through Cation Substitution in Ultra-Broadband Near-Infrared Phosphors.

The growing interest in the use of near-infrared (NIR) radiation for spectroscopy, optical communication, and medical applications spanning both NIR-I (700-900 nm) and NIR-II (900-1700 nm) has driven the need for new NIR light sources. NIR phosphor-converted light-emitting diodes (pc-LEDs) are expected to replace traditional lamps mainly due to their high efficiency and compact design. Broadband NIR phosphors activated by Cr3+ and Cr4+ have attracted significant research interest, offering emission across a wide range from 700 to 1700 nm. In this work, we synthesized a series of Sc2(1-x)Ga2xO3:Cr3+/4+ materials (x = 0-0.2) with broadband NIR-I (Cr3+) and NIR-II (Cr4+) emission. We observed a substantial increase in the intensity of Cr3+ (approximately 77 times) by incorporating Ga3+ ions. Additionally, our investigation revealed that energy transfer occurred between Cr3+ and Cr4+ ions. Configuration diagrams are presented to elucidate the behavior of Cr3+ and Cr4+ ions within the Sc2O3 matrix. We also observed a phase transition at a pressure of 20.2 GPa, resulting in a new unknown phase where Cr3+ luminescence exhibited a high-symmetry environment. Notably, this study presents the pressure-induced shift of NIR Cr4+ luminescence in Sc2(1-x)Ga2xO3:Cr3+/4+. The linear shifts were estimated at 83 ± 3 and 61 ± 6 cm-1/GPa before and after the phase transition. Overall, our findings shed light on the synthesis, luminescent properties, temperature, and high-pressure behavior within the Sc2(1-x)Ga2xO3:Cr3+/4+ materials. This research contributes to the understanding and potential applications of these materials in the development of efficient NIR light sources and other optical devices.

A parametric bootstrap approach for computing confidence intervals for genetic correlations with application to genetically-determined protein-protein networks.

Genetic correlation refers to the correlation between genetic determinants of a pair of traits. When using individual-level data, it is typically estimated based on a bivariate model specification where the correlation between the two variables is identifiable and can be estimated from a covariance model that incorporates the genetic relationship between individuals, e.g., using a pre-specified kinship matrix. Inference relying on asymptotic normality of the genetic correlation parameter estimates may be inaccurate when the sample size is low, when the genetic correlation is close to the boundary of the parameter space, and when the heritability of at least one of the traits is low. We address this problem by developing a parametric bootstrap procedure to construct confidence intervals for genetic correlation estimates. The procedure simulates paired traits under a range of heritability and genetic correlation parameters, and it uses the population structure encapsulated by the kinship matrix. Heritabilities and genetic correlations are estimated using the close-form, method of moment, Haseman-Elston regression estimators. The proposed parametric bootstrap procedure is especially useful when genetic correlations are computed on pairs of thousands of traits measured on the same exact set of individuals. We demonstrate the parametric bootstrap approach on a proteomics dataset from the Jackson Heart Study.

Inflammatory Biomarkers and Blood Physical Property Transformations Following On-Pump Coronary Artery Bypass Graft Surgery.

OBJECTIVE: This study aimed to compare the hemorheological and inflammatory changes before and after coronary artery bypass graft (CABG) surgery, as factors such as hypothermia, hemodilution, transfusion, and other variables affect blood viscosity and inflammation during the procedure. METHODS: A total of 25 patients who underwent CABG surgery were enrolled in this study. Whole blood was collected just before the CABG (D0), 2 days after surgery (D2), and 5 days after surgery (D5). The plasma viscosity (PV) and whole blood viscosity (WBV) were measured at shear rates ranging from 0.1 to 1000 s-1 using a rheometer, and the mean values were compared. Inflammatory markers were also assessed and analyzed in relation to the hemorheological changes. RESULTS: Compared with the baseline values, the PV significantly increased after 5 days. WBV showed a significant increase on day 2 and after 5 days. The WBV and fibrinogen were significantly correlated on day 2 and day 5 but not before surgery. Inflammatory markers such as CRP, WBC, platelets, and fibrinogen also demonstrated notable changes in relation to the hemorheological alterations. CONCLUSIONS: This study highlights the crucial finding that hyperviscosity, characterized by elevated PV and WBV, persists for almost one week after on-pump CABG surgery. Understanding the interplay between inflammation and hemorheological properties during the postoperative period is crucial for optimizing patient care. Future research should focus on exploring the underlying mechanisms and potential therapeutic interventions to mitigate the impact of inflammation on blood viscosity and improve patient outcomes following CABG surgery.

Chain-Kinked Design: Improving Stretchability of Polymer Semiconductors through Nonlinear Conjugated Linkers.

The manipulation of the polymer backbone structure has a profound influence on the crystalline behavior and charge transport characteristics of polymers. These strategies are commonly employed to optimize the performance of stretchable polymer semiconductors. However, a universal method that can be applied to conjugated polymers with different donor-acceptor combinations is still lacking. In this study, we propose a universal strategy to boost the stretchability of polymers by incorporating the nonlinear conjugated linker (NCL) into the main chain. Specifically, we incorporate meta-dibromobenzene (MB), characterized by its asymmetric linkage sites, as the NCL into the backbone of diketopyrrolopyrrole-thiophene-based (DPP-based) polymers. Our research demonstrates that the introduction of MB prompts chain-kinking, thereby disrupting the linearity and central symmetry of the DPP conjugated backbone. This modification reshapes the polymer conformation, decreasing the radius of gyration and broadening the free volume, which consequently adjusts the level of crystallinity, leading to a considerable increase in the stretchability of the polymer. Importantly, this method increases stretchability without compromising mobility and exhibits broad applicability across a wide range of donor-acceptor pair polymers. Leveraging this strategy, fully stretchable transistors were fabricated using a DPP polymer that incorporates 10 mol % of MB. These transistors display a mobility of approximately 0.5 cm2 V-1 s-1 and prove remarkably durable, maintaining 90% of this mobility even after enduring 1000 cycles at 25% strain. Overall, we propose a method to systematically control the main-chain conformation, thereby enhancing the stretchability of conjugated polymers in a widely applicable manner.

Hidden Hexavalent Chromium Ions with Subtle Structural Evolution in Near-Infrared Phosphors.

Cr-doped inorganic materials are pivotal in developing near-infrared optical materials; however, multivalent Cr ions and their respective distribution in the materials remain ambiguous. Herein, a series of Li(Sc1-xInx)O2:Cr phosphors containing both Cr3+/Cr6+ ions are prepared. High-resolution synchrotron X-ray diffraction (XRD) reveals two similar phases in Li(Sc1-xInx)O2. Raman spectra further confirm distinct scattering patterns for the two end-member compositions, corroborating the findings from the synchrotron XRD analysis. Cr K-edge X-ray absorption near-edge structure and extended X-ray absorption fine structure demonstrate that most Cr ions in the as-prepared samples are Cr6+, while Cr3+ becomes dominant after washing with water. Moreover, the source and distribution of Cr3+ and Cr6+ ions in the as-prepared and washed samples are revealed through X-ray fluorescence and X-ray excited optical luminescence techniques, which indicate that Cr6+ ions aggregate within the sample, while Cr3+ ions are evenly distributed. Photoluminescence, decay curves, and line shape analyses are implemented to resolve the electron-lattice interactions, and the corresponding mechanisms are provided to explain the asymmetry between photoluminescence and photoluminescence excitation spectra. Overall, this study provides valuable insights into the distribution of low-concentration multivalence ions in solid-state materials and offers a deeper understanding of the approaches to precisely resolve the subtle changes in the crystal structure.

Interpretable speech features vs. DNN embeddings: What to use in the automatic assessment of Parkinson's disease in multi-lingual scenarios.

Speech-based approaches for assessing Parkinson's Disease (PD) often rely on feature extraction for automatic classification or detection. While many studies prioritize accuracy by using non-interpretable embeddings from Deep Neural Networks, this work aims to explore the predictive capabilities and language robustness of both feature types in a systematic fashion. As interpretable features, prosodic, linguistic, and cognitive descriptors were adopted, while x-vectors, Wav2Vec 2.0, HuBERT, and TRILLsson representations were used as non-interpretable features. Mono-lingual, multi-lingual, and cross-lingual machine learning experiments were conducted leveraging six data sets comprising speech recordings from various languages: American English, Castilian Spanish, Colombian Spanish, Italian, German, and Czech. For interpretable feature-based models, the mean of the best F1-scores obtained from each language was 81% in mono-lingual, 81% in multi-lingual, and 71% in cross-lingual experiments. For non-interpretable feature-based models, instead, they were 85% in mono-lingual, 88% in multi-lingual, and 79% in cross-lingual experiments. Firstly, models based on non-interpretable features outperformed interpretable ones, especially in cross-lingual experiments. Specifically, TRILLsson provided the most stable and accurate results across tasks and data sets. Conversely, the two types of features adopted showed some level of language robustness in multi-lingual and cross-lingual experiments. Overall, these results suggest that interpretable feature-based models can be used by clinicians to evaluate the deterioration of the speech of patients with PD, while non-interpretable feature-based models can be leveraged to achieve higher detection accuracy.

Multiresponsive Self-Healing Lanthanide Fluorescent Hydrogel for Smart Textiles.

Lanthanide organometallic complexes exhibit strong luminescence characteristics, owing to their antenna effects. The f-d energy level transition causes this phenomenon, which occurs when ligands and the external electrons of lanthanide metals coordinate. Based on this phenomenon, we used two lanthanide metals, europium (Eu) and terbium (Tb), in the present study as the metal center for iminodiacetic acid ligands. Further, we developed the resulting fluorescent organometallic complex as a smart material. The ligand-metal bond in the material functioned as a metal chelating agent and a cross-linking agent in a dynamically coordinated form, thereby prompting the material to self-heal. Temperature-sensitive poly-N-isopropylacrylamide was incorporated into the material as the polymer backbone. Afterward, we combined it with water-soluble poly(vinyl alcohol) and an additional ligand from poly(acrylic acid) to fabricate a high-performance hydrogel composite material. The shrinkage and expansion of the polymer form a grid between the materials. Because of the different coordination stabilities of Eu3+ and Tb3+, the corresponding material exhibits environmental responses toward excitation wavelength, temperature, and pH, thus generating different colors. When used in fabrics, the cross-linking mechanism of the material effectively looped the material between fabric fibers; furthermore, the temperature sensitivity of the polymer adjusted the size of pores between fabric fibers. At relatively higher temperatures (>32 °C), the polymer structure shrank, fiber pores expanded, and air permeability improved. Thus, this material appears to be promising for use in smart textiles.

A common East-Asian ALDH2 mutation causes metabolic disorders and the therapeutic effect of ALDH2 activators.

Obesity and type 2 diabetes have reached pandemic proportion. ALDH2 (acetaldehyde dehydrogenase 2, mitochondrial) is the key metabolizing enzyme of acetaldehyde and other toxic aldehydes, such as 4-hydroxynonenal. A missense Glu504Lys mutation of the ALDH2 gene is prevalent in 560 million East Asians, resulting in reduced ALDH2 enzymatic activity. We find that male Aldh2 knock-in mice mimicking human Glu504Lys mutation were prone to develop diet-induced obesity, glucose intolerance, insulin resistance, and fatty liver due to reduced adaptive thermogenesis and energy expenditure. We find reduced activity of ALDH2 of the brown adipose tissue from the male Aldh2 homozygous knock-in mice. Proteomic analyses of the brown adipose tissue from the male Aldh2 knock-in mice identifies increased 4-hydroxynonenal-adducted proteins involved in mitochondrial fatty acid oxidation and electron transport chain, leading to markedly decreased fatty acid oxidation rate and mitochondrial respiration of brown adipose tissue, which is essential for adaptive thermogenesis and energy expenditure. AD-9308 is a water-soluble, potent, and highly selective ALDH2 activator. AD-9308 treatment ameliorates diet-induced obesity and fatty liver, and improves glucose homeostasis in both male Aldh2 wild-type and knock-in mice. Our data highlight the therapeutic potential of reducing toxic aldehyde levels by activating ALDH2 for metabolic diseases.

Risks after Gestational Diabetes Mellitus in Taiwanese Women: A Nationwide Retrospective Cohort Study.

OBJECTIVE: An increasing trend in the prevalence of gestational diabetes mellitus (GDM) has been reported in Taiwan. GDM has been linked to various adverse maternal outcomes over a long period, including cardiovascular disease (CVD) and chronic kidney disease (CKD). However, evidence implies that the effects of GDM on the mid-term surrogate risk factors for these diseases are limited. Furthermore, data from nationwide cohort studies are limited. The primary aim of this study was to investigate the risk of developing type 2 diabetes mellitus (T2DM), arterial hypertension (aHTN), and hyperlipidemia (HL) through a 5-year follow-up post-delivery of women with GDM in a nationwide cohort study in Taiwan. The second objective was to investigate the risk of developing insulin resistance syndrome (IRS)-related diseases, including CVD, acute myocardial infarction (AMI), peripheral artery occlusive disease (PAOD), non-alcoholic fatty liver diseases (NAFLD), and CKD. METHODS: This was a retrospective, population-based nationwide cohort study. The data source comprises a merge of the Birth Certificate Application Database (BCA) and the National Health Insurance Research Database in Taiwan. Women aged between 15 and 45 years who gave birth in Taiwan between 2004 and 2011 were included. Women who were enrolled and had a GDM diagnosis were assigned to the exposure group. Women who were enrolled without a GDM diagnosis were assigned to the comparison group. The relative risk of developing T2DM, aHTN, HL, and IRS-related diseases, including CVD, AMI, PAOD, NAFLD, and CKD, were analyzed and presented as hazard ratio (HR) through Cox regression and log-rank regression analyses. RESULTS: A total of 1,180,477 women were identified through the BCA database between 2004 and 2011. Of those, 71,611 GDM-diagnosed women and 286,444 women without GDM were included in the final analysis. After adjusting for age, pre-existing cancer, and parity, developing T2DM, aHTN, and HL were still significantly increased in the GDM group (HR and interquartile range (IQR): 2.83 (2.59, 3.08), 1.09 (1.01, 1.06), and 1.29 (1.20, 1.38), accordingly). CVD, NAFLD, and CKD had a very low incidence and showed insignificant results. CONCLUSION: Our findings provide nationwide cohort data showing that GDM increased the risk of developing T2DM, aHTN, and HL 5 years after delivery within the same group. The GDM complications and risk of CVD, AMI, PAOD, NAFLD, and CKD need further investigation.

Rege-1 promotes C. elegans survival by modulating IIS and TOR pathways.

Metabolic pathways are known to sense the environmental stimuli and result in physiological adjustments. The responding processes need to be tightly controlled. Here, we show that upon encountering P. aeruginosa, C. elegans upregulate the transcription factor ets-4, but this upregulation is attenuated by the ribonuclease, rege-1. As such, mutants with defective REGE-1 ribonuclease activity undergo ets-4-dependent early death upon challenge with P. aeruginosa. Furthermore, mRNA-seq analysis revealed associated global changes in two key metabolic pathways, the IIS (insulin/IGF signaling) and TOR (target of rapamycin) kinase signaling pathways. In particular, failure to degrade ets-4 mRNA in activity-defective rege-1 mutants resulted in upregulation of class II longevity genes, which are suppressed during longevity, and activation of TORC1 kinase signaling pathway. Genetic inhibition of either pathway way was sufficient to abolish the poor survival phenotype in rege-1 worms. Further analysis of ETS-4 ChIP data from ENCODE and characterization of one upregulated class II gene, ins-7, support that the Class II genes are activated by ETS-4. Interestingly, deleting an upregulated Class II gene, acox-1.5, a peroxisome ß-oxidation enzyme, largely rescues the fat lost phenotype and survival difference between rege-1 mutants and wild-types. Thus, rege-1 appears to be crucial for animal survival due to its tight regulation of physiological responses to environmental stimuli. This function is reminiscent of its mammalian ortholog, Regnase-1, which modulates the intestinal mTORC1 signaling pathway.

Prime Editing Strategy to Install the PRPH2 c.828+1G>A Mutation.

Mutations in peripherin 2 (PRPH2) are associated with a spectrum of inherited retinal diseases (IRDs) including retinitis pigmentosa (RP) and macular degeneration. As PRPH2 is localized to cone and rod outer segments, mutations in PRPH2 lead the disorganization or absence of photoreceptor outer segments. Here, we report on a patient with PRPH2-linked RP who exhibited widespread RPE atrophy with a central area of macular atrophy sparing the fovea. In future studies, we plan to model the pathobiology of PRPH2-based RP using induced pluripotent stem cell (iPSC)-derived retinal organoids. To effectively model rare mutations using iPSC-derived retinal organoids, we first require a strategy that can install the desired mutation in healthy wild-type iPSC, which can efficiently generate well-laminated retinal organoids. In this study, we developed an efficient prime editing strategy for the installation of the pathogenic PRPH2 c.828+1 G>A splice-site mutation underlying our patient's disease.

Generation of an Avian Myeloblastosis Virus (AMV) Reverse Transcriptase Prime Editor.

Prime editing (PE) is a novel, double-strand break (DSB)-independent gene editing technology that represents an exciting avenue for the treatment of inherited retinal diseases (IRDs). Given the extensive and heterogenous nature of the 280 genes associated with IRDs, genome editing has presented countless complications. However, recent advances in genome editing technologies have identified PE to have tremendous potential, with the capability to ameliorate small deletions and insertions in addition to all twelve possible transition and transversion mutations. The current PE system is based on the fusion of the Streptococcus pyogenes Cas9 (SpCas9) nickase H840A mutant and an optimized Moloney murine leukemia virus (MMLV) reverse-transcriptase (RT) in conjunction with a PE guide RNA (pegRNA). In this study, we developed a prime editor based on the avian myeloblastosis virus (AMV)-RT and showed its applicability for the installation of the PRPH2 c.828+1G>A mutation in HEK293 cells.